News

Results - Run 61, B31, H19

Run 61 (General module), B31 (Breast module) and H19 (HER2 ISH) were accomplished from January to April 2021. At present >550 participants from more than 60 different countries are registered in one or more of the modules offered.

The number of laboratories returning slides in due time for the assessment sessions decreased in this run 61 compared to previous assessments, and mainly caused to the COVID-19 pandemic and associated postal delays. All slides returned after the assessment were assessed and received advice if the result being insufficient but were not be included in the individual reports.

A short summary of the results is given below. Click on the epitope name to see the complete and detailed assessment report for the individual markers evaluated. In each assessment report, NordiQC intended to identify best practice methods and reliable and recommendable immunohistochemical critical assay performance controls (iCAPCs) to both guide the IHC protocol set-up and to monitor the IHC performance within the laboratories. 

General module - run 61

CD15: 305 laboratories participated and obtained a pass rate of 86% (53% optimal) being the highest level in all six assessments performed in NordiQC. 100% of insufficient results were characterized by generally too weak or false negative results for CD15.   

CD56: 324 laboratories participated and obtained a pass rate of 62% (36% optimal). The pass rate was significantly reduced compared to the latest assessment, run 37 2013. In this assessment, the rmAb clone MRQ-42 provided superior performance with a total pass rate of 99% (136/137) for both concentrated and RTU formats. This clone was very robust and provided a high proportion of optimal results on all main IHC platforms and by several different protocol settings. Due to the low pass rate, CD56 will be repeated in 2022, run 64 and thus no reassessment will be offered.

CDX2: 325 laboratories participated and obtained a pass rate of 91% (79% optimal) being the highest level in all six assessments performed in NordiQC. The extended use of robust and successful Abs as rmAb clone EPR2765Y, access to optimally performing RTU systems from the main IHC system vendors and identification of pancreas as iCAPC seem to contribute to the improvement.  

MSH6: 278 laboratories participated and obtained a pass rate of 91% (79% optimal) and a significant improvement from run 52, 2018 with a pass rate of 52%. The access to several high quality RTU systems for MSH6 was instrumental for the improved pass rate. Especially the recently introduced RTU system of rmAb clone SP93 (Ventana/Roche) showed a significantly superior performance compared to the terminated system based on clone 44, being widely used in run 52.

p63: 324 laboratories participated and obtained a pass rate of 79% (37% optimal). 97% of insufficient results were characterized by generally too weak or false negative results for p63. Efficient HIER preferable in combination with use of 3-step detection systems were main prerequisites for optimal p63 results. The widely use mAb clones DAK-p63 and 4A4 were significantly more successful compared to clone 7JUL.

Breast module - run B31

Estrogen receptor (ER): 378 laboratories participated and obtained a pass rate of 85% (48% optimal). rmAb clones SP1 and EP1 and mAb clone 6F11 could all be used to provide an optimal result. 86% of the participants used RTU systems for the demonstration of ER. The RTU systems from Ventana/Roche and Dako/Agilent for BenchMark and Omnis respectively, were most successful with pass rates of 93 and 96%. Too weak or false negative staining reactions were the predominant features of insufficient results. Tonsil was found to be recommendable as positive and negative tissue control.

Progesterone receptor (PR): 377 laboratories participated and obtained a pass rate of 92% (68% optimal). mAb clones 16, PgR 636, PgR 1294 and rmAb clone 1E2 could all be used to provide an optimal result. 77% of the participants used RTU systems for the demonstration of PR. The RTU systems from Ventana/Roche, Dako/Agilent and Leica Biosystems used as “plug-and-play” assays provided an overall pass rate of 99%. In this assessment, the insufficient results were mainly characterized by false negative results. Tonsil and uterine cervix in combination were found to be recommendable negative and positive tissue controls. 

HER2 IHC: 362 laboratories participated and obtained a pass rate of 92% (75% optimal). In this assessment, the CE-IVD approved HER2 IHC assays PATHWAY®, 4B5 (Ventana/Roche) and the recently launched HercepTest™ Dako/Agilent for Omnis were most successful providing a pass rate of 100%. LDTs based on a concentrate or RTU with no predictive claim were used by 24% giving a pass rate of 81% (39% optimal). The insufficient results were primarily characterized by false negative results or a poor signal-to-noise ratio compromising the interpretation.   

HER2 ISH module - run H19

HER2 ISH: 143 laboratories participated in the technical assessment of HER2 ISH (BRISH) and obtained a pass rate of 70% (41% optimal) similar to the level obtained in most recent assessments. The VENTANA HER2 Dual ISH DNA Probe Cocktail, 800-6043 and INFORM™ HER2 Dual ISH assay, 800-4422/780-4422 were most widely used with pass rates of 75% and 64%, respectively. Insufficient results were mainly caused by large negative areas and missing signals.

Laboratories performing FISH (n=65) achieved a slightly higher consensus rate for the interpretation of HER2 amplification status compared to laboratories performing BRISH.

 

April 20th 2021,
Søren Nielsen
Scheme director

Results - Run 60, B30, H18, C8

Run 60 (General module), B30 (Breast module), C8 (Companion modules) and H18 (HER2 ISH) module were completed from October to December 2020. In total, 485 laboratories participated in one or more of the modules offered.

Similar to the spring assessment, shipping of slides to and from the participants was challenged by the Covid-19 pandemic causing delays. Consequently, the overall participation rates were slightly reduced compared to normal levels. All slides returned after the specified assessment dates were assessed but not included in the general reports. The composition of assessor teams was also affected by travel restrictions limiting the possibility to include international assessors. Despite the challenge, all assessor teams were anchored on representation of both IHC expert pathologists and biomedical scientists primarily from Denmark. NordiQC is proud and grateful for the extensive support from our assessors and willingness to secure our core functions in such critical situations. 

As described previously in news on 7th October, we were notified by some participants observing aberrant IHC results on slides circulated for especially p40 and PAX8. Completely false negative or uneven staining reactions were reported. This was most likely caused by quality issues on some of the SuperFrost Plus slides preventing reagents to spread. The issue was not seen by the internal validation process by NordiQC for any of the markers offered and in total during the assessments about 70 slides out of more than 2.600 slides assessed potentially were affected by technical quality of the slides circulated. For each assessment, slides characterized by uneven staining or a completely false negative result that might be related to the quality of the slide and not the protocol submitted, was given a comment in the individual assessment feed-back. We are in the process of evaluating if other glass types are superior to SuperFrost Plus slides for our EQA program with particular focus on the most common IHC platforms used by our participants, and we will circulate slides based on these findings in 2021.

As usual, a short summary of the results is given below. Click on the epitope name to see the complete and detailed assessment report for the markers evaluated. In each assessment report, NordiQC intended to identify best practice methods and reliable and recommendable immunohistochemical critical assay performance controls (iCAPCs) to both guide the IHC protocol set-up and to monitor the IHC performance within the laboratories.

General module - run 60

CD68: 329 laboratories participated and obtained a pass rate of 83% (32 % optimal). The mAb clones PG-M1, KP1, 514H12 and rmAb clone GR021 could all be used to provide an optimal result for CD68. PG-M1 was most widely used and gave the highest proportion of optimal results both as concentrate within a laboratory developed (LD) assay and as Ready-To-Use (RTU) system. The Intended purpose of the assessment focused on general pathology to identify macrophages and lung pathology to differentiate macrophages from epithelial tumour cells as support for accurate read-out of PD-L1 TPS.

MLA: 312 laboratories participated and obtained a pass rate of 88% (26% optimal) being the highest level seen in the NordiQC runs performed. The assessment focused on the use of MLA in the diagnosis of melanoma. The mAb clone A103 was the most widely used antibody for demonstration of MLA and applied by 88% of the laboratories. The pass rate was high both as concentrated format and as RTU system, but the level of optimal results was low at 18%. The relatively recently launched rmAb clone EP43, was most successful with a pass rate of 100%, 97% optimal.

p40: 262 laboratories participated.  This was the third assessment of p40 in NordiQC and a pass rate of 86% was obtained, which is a significant increase compared to the result obtained in the previous runs. The increased pass rate may be explained by an extended use of the highly sensitive and robust mAb clone BC28 both as concentrate and as RTU format. Additionally, the proportion of laboratories using less successful polyclonal antibodies has been reduced from 41% in run 44 to 7% in this run. The identification of tonsil and placenta as reliable iCAPCS has facilitated the process to calibrate and monitor the performance of IHC assays for p40.

PAX8: 271 laboratories participated and obtained a pass rate of 39% (11% optimal). The pass rate has thus in two successive runs been at a very low level. The low pass rate was especially related to extended use of the less successful mAb clone MRQ-50 providing a poor performance on the Ventana BenchMark and Dako Omnis platforms and in addition cross-reaction with PAX5 in B-cells. In contrast, the rmAbs clones EP298 and SP348 gave encouraging results and a high proportion of sufficient results on the main fully automated platforms and no cross-reaction with e.g. PAX5 was observed. Due to the low pass rate in the two runs, PAX8 will be repeated in run 62, 2021 and consequently no re-assessment offered for run 60.

SOX10: 250 laboratories participated and obtained a pass rate of 92% (67% optimal) being the highest level seen in the four NordiQC runs performed. The assessment focused on the use of SOX10 identifying malignant melanomas and triple negative breast carcinoma in the characterization of tumours of unknown origin. The Ventana RTU system based on the rmAb clone SP267 for BenchMark was most successful providing a pass rate of 98%, 95% optimal when applied by recommended protocol settings.  

Breast module - run B30

ER: 363 laboratories participated and obtained a pass rate of 94% (58% optimal). rmAb clones SP1 and EP1 and mAb clone 6F11 could all be used to provide an optimal result. 86% of the participants used RTU systems for the demonstration of ER. In this assessment, low analytical sensitivity giving a too weak or false negative staining reaction was the predominant feature of insufficient results. Tonsil was found to be recommendable as positive and negative tissue control with particular possibility to ensure an appropriate low limit of demonstration of ER – concordant to the ASCO/CAP 2020 recommendations on ER IHC testing.

HER2 IHC: 343 laboratories participated and obtained a pass rate of 92% (63% optimal). In this assessment, the FDA-/CE-IVD approved HER2 IHC assay PATHWAY® /4B5 Ventana/Roche was the most accurate and successful assays providing a pass rate of 99% and 97%, respectively. LD assays based on concentrated Abs or generic RTU Abs without predictive claim were used by 25% and for this group a pass rate of 80% was obtained. The insufficient results primarily characterized by false negative results or a poor signal-to-noise ratio compromising the interpretation.   

HER2 ISH module - run H18

HER2 ISH: 141 laboratories participated in the technical assessment of HER2 ISH and obtained a pass rate of 69% (40% optimal). In this assessment an increased number of participants used the VENTANA HER2 Dual ISH DNA Probe Cocktail, 800-6043 providing a pass rate of 80% on the expense of the less successful INFORM™ HER2 Dual ISH assay, 800-4422/780-4422 with a pass rate of only 38%. In total 94% of the participants used one of these two, Ventana ISH systems. The insufficient results were mainly characterized by large negative areas and missing signals. 61 laboratories submitted a FISH result and participated in the interpretation segment. Laboratories performing FISH achieved a slightly higher consensus rate for the interpretation of HER2 amplification status compared to laboratories performing BRISH.

Companion module - run C8

PD-L1 KEYTRUDA: 200 laboratories participated and obtained a pass rate of 86% (49% optimal). This assessment for PD-L1 KEYTRUDA® primarily focused on the evaluation of the analytical accuracy of the IHC assays to identify patients with NSCLCs and urothelial carcinomas to be treated with KEYTRUDA® as immune therapy. Similar to observations seen in previous runs, the insufficient PD-L1 IHC results were most frequently characterized by a reduced proportion of PD-L1 positive cells compared to the level expected and defined by the NordiQC reference standard methods. This resulted in a too low TPS/CPS changing PD-L1 status in one or more of the carcinomas included. The companion diagnostic PD-L1 IHC assays, 22C3 GE006, Dako/Agilent and SP263 741-4905, Ventana/Roche performed in concordance to the product guidelines, were most successful providing a high proportion of sufficient and optimal results and superior to LD assays.

PD-L1 TECENTRIQ: 123 laboratories participated and obtained a pass rate of 63% (33% optimal), being slightly improved compared to run C7 with a pass rate of 55%. The assessment for PD-L1 TECENTRIQ® primarily focused on evaluation of the analytical accuracy of the PD-L1 IHC assays to identify patients with urothelial carcinomas or triple negative breast carcinomas (TNBC) to be treated with TECENTRIQ® as immune therapy. The PD-L1 SP142 CDx IHC assay product no. 741-4860 and the IHC assay 790-4860 from Ventana/Roche were the most successful assays. Other PD-L1 CDx assays as SP263 (741-4905, Ventana) and 22C3 (SK006/GE006, DakoAgilent) being very successful in the NordiQC PD-L1 KEYTRUDA® assessments provided no sufficient staining results. The insufficient results were typically characterized by a too strong staining reaction in tumour cells in one or more of the carcinomas compromising the interpretation of PD-L1 reaction in immune cells.

 

December 14th, 2020

Søren Nielsen,
Scheme director

 

Important updates on run 60, B30, H17 and C8

Run 60 (General module), B30 (Breast module), C8 (Companion module) and H17 (HER2 ISH) is now being executed.

Due to the Covid-19 pandemic and transportation challenges for postal services, we anticipate delays for slides to be returned to NordiQC for the official assessment dates. We will do our very best to assess all slides returned, but due to the extensive data analysis, only slides received and available for the assessment of the individual modules will be integrated in the assessment reports.

We have been informed by some laboratories, that slides for p40 and PAX8 have revealed aberrant properties producing uneven staining and/or completely negative results. During the internal NordiQC validation of the circulated material, we did not observe any issues and subsequent retests have not provided any transparency of the problem. However, the problems are most likely related to change in the hydrophilic character of the SuperFrost Plus slides to be more hydrophobic and hereby preventing reagents to spread. We are now in the process to validate new slides for our EQA program and hopefully will identify slides with improved consistency.

In this assessment, slides returned to NordiQC with clear indication of aberrant results caused by the slide properties will be marked as “technically insufficient” and not assessed. We will shortly after the assessment explore the option to resend new slides to the laboratories being affected by this issue. We apologize for any inconvenience.

 

Søren Nielsen Scheme director

Results - Run 59, C7

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 Run 59 (General module) and C7 (Companion modules) were accomplished from April to July 2020. In total, 366 laboratories participated in one or more of the modules offered. 

The assessments were affected by the Covid-19 pandemic and especially the shipping of slides to and from the participants was challenged. The overall participation rates were reduced to 80-90% compared to 95-98%, normally seen. However, for all markers adequate number of laboratories and data points were accomplished to generate assessment reports and conclusions. All slides returned will be assessed but not be included in the online available reports.        

As usual, a short summary of the results is given below. Click on the epitope name to see the complete and detailed assessment report for the individual markers evaluated. In each assessment report, NordiQC intended to identify best practice methods and reliable and recommendable immunohistochemical critical assay performance controls (iCAPCs) to both guide the IHC protocol set-up and to monitor the IHC performance within the laboratories.

General module - run 59

ASMA: 260 laboratories participated and obtained a pass rate of 69% (28 % optimal) being the highest level in all six NordiQC assessment runs for ASMA. The Ready-To-Use (RTU) systems from Dako and Leica based on the mAb clones 1A4 and asm-1, respectively were most successful. The mAb clone 1A4 provided an inferior performance on the Ventana BenchMark platforms both as concentrate within a laboratory developed (LD) assay and as Ready-To-Use (RTU) system. The recently launched mAb clone BS66 showing promising performance in previous assessments and used within the NordiQC reference standard methods for ASMA, was less successful giving unreproducible and unspecific results. The product was during the assessment terminated by the vendor due to lot-to-lot inconsistency.

CD10: 295 laboratories participated and obtained a pass rate of 79% (57% optimal). The assessment focused on CD10 as part on an IHC panel discriminating Diffuse Large B-Cell Lymphoma (DLBCL) of Germinal centre B-cell like (GCB) from non-GCB subtype. The well-established and widely used mAb clone 56C6 was most successful both within a LD assay and especially as RTU system from Dako and Leica. HIER in alkaline buffer, precise calibration of the primary Ab and use of a 3-step polymer or multimer based detection system were the main prerequisites for an optimal result.  

CD45: 296 laboratories participated.  This was the third assessment of CD45 in NordiQC and a pass rate of 94% was obtained, which is a significant increase compared to the result obtained in the previous runs. The use of robust primary Abs (e.g. mAb clones 2B11+PD7/26) but also well calibrated RTU systems accounted for the superior performance obtained in this assessment. The identification of tonsil and liver as reliable iCAPCS has facilitated the process to calibrate and monitor the performance of IHC assays for CD45. 

p16: 291 laboratories participated and obtained a pass rate of 83% (50% optimal). This was the second assessment of p16 in NordiQC and the pass rate increased from 70% seen in run 26, 2009. The assessment focused on p16 identifying HPV associated cervical lesions in histological uterine cervical samples. 78% of the participants used the established Ventana CINtec® RTU products based on mAb clone E6H4 giving a pass rate of 100% when applied as “Plug-and-Play” system. In 2019 and 2020 many new Abs have been launched and especially the mAb clone MX007 was found successful. The mAb clone G175-405 showed an inferior and unsatisfactory performance.  

S100: 299 laboratories participated and obtained a pass rate of 82% (48% optimal) and identical to the pass rate in the recent run 50, 2017.  The assessment focused on S100 used to identify the origin of melanomas of cancer of unknown primary (CUP) and differentiate schwannoma from other spindle cell lesions as leiomyoma. In general, pAbs were more successful than mAbs both within LD assays based on concentrated formats or as RTU systems. Especially the Dako RTU systems provided a high pass rate and proportion of optimal results. An increased number of participants use RTU systems, 58% in this run compared to 46% in run 50, most likely caused by the “gold-standard” S100 pAb Z0311, Dako not being commercially available anymore.   

URO II/III: 66 laboratories participated and obtained a pass rate of 45% (21% optimal). This was the first assessment of URO II/III in NordiQC and focused on identification of urothelial origin of cancer of unknown primary (CUP). The relatively low pass rate was especially related to extended use of less successful Abs and in particular related to Abs against URO III. In contrast the mAb clone BC21 was very successful and in total used by 47% of the participants either as concentrated format or as RTU giving an overall pass rate of 87% (45% optimal). Urethra and tonsil in combination seem appropriate as central positive and negative tissue controls to calibrate and monitor the performance of IHC assay for URO II/III.

Companion module - run C7

PD-L1 KEYTRUDA®: 191 laboratories participated and obtained a pass rate of 83% (55% optimal). This assessment for PD-L1 KEYTRUDA® primarily focused on the evaluation of the analytical accuracy of the IHC assays to identify patients with NSCLCs and urothelial carcinomas to be treated with KEYTRUDA® as immune therapy. Similar to the observations seen in previous runs, the insufficient PD-L1 IHC staining results were most frequently characterized by a reduced proportion of PD-L1 positive cells compared to the level expected and defined by the PD-L1 IHC pharmDx assay, SK006 (Dako/Agilent). This resulted in a too low TPS/CPS changing the PD-L1 status in one or more of the carcinomas included in the assessment material. The companion diagnostic PD-L1 IHC assays, 22C3 GE006, Dako and SP263 741-4905, Ventana performed in concordance to the product guidelines, were most successful providing a high proportion of sufficient and optimal results and superior to LD assays.

PD-L1 TECENTRIQ®: 102 laboratories participated and obtained a pass rate of 55% (30% optimal), being significantly reduced from the results in the first run C7 with a pass rate of 76%. The assessment for PD-L1 TECENTRIQ® primarily focused on evaluation of the analytical accuracy of the PD-L1 IHC assays to identify patients with urothelial carcinomas or triple negative breast carcinomas (TNBC) to be treated with TECENTRIQ® as immune therapy. In concordance to run C6, the PD-L1 SP142 companion diagnostic IHC assay 741-4860 from Ventana was most successful providing a pass rate of 93%. Other companion diagnostic and LD assays provided significantly lower pass rates - grouped together 18%.  insufficient results were most frequently characterized by a reduced proportion or too weak specific staining reaction of immune cells combined with an excessive staining reaction of tumour cells compromising the scoring and PD-L1 status in the immune cells.

July 10th 2020
Søren Nielsen, Scheme director

Results - Run 58, B29, H17

Run 58 (General module), B29 (Breast module) and H17 (HER2 ISH) were accomplished from January to April 2020. In total 448 laboratories participated in one or more of the modules offered. 

A short summary of the results is given below. Click on the epitope name to see the complete and detailed assessment report for the individual markers evaluated. In each assessment report, NordiQC intended to identify best practice methods and reliable and recommendable immunohistochemical critical assay performance controls (iCAPCs) to both guide the IHC protocol set-up and to monitor the IHC performance within the laboratories. 

Please note, table 1 has been adjusted. The table now displays the percentage of sufficient and optimal results for a specific antibody (last two rows right). Due to the expanded use of antibodies for Ready-To-Use systems, table 1 now shows the performance of these systems when used by vendor recommended protocol settings (VRPS) or by laboratory modified protocol settings (LMPS).

General module - run 58

CK-PAN: 326 laboratories participated and obtained a pass rate of 75% (52% optimal). The pass rate and performance was highly influenced by choice of the antibody (Ab),  retrieval method and IHC platform used. The mAb clone cocktail AE1/AE3 provided a high pass rate and proportion of optimal results when used with HIER both as concentrate within a laboratory developed (LD) assay and as Ready-To-Use (RTU) system assay, whereas the clone cocktail AE1/AE3/PCK26 required a combined pre-treatment with HIER and gentle proteolysis for optimal performance. The clones MNF116 and Oscar showed an inferior performance and cannot be recommended as CK-PAN primary Ab.    

TTF1: 322 laboratories participated and obtained a pass rate of 80% (56% optimal) being the highest level in all seven NordiQC assessment runs for TTF1. In concordance with previous NordiQC assessments for TTF1, the mAb clone SPT24 and the rmAb clone SP141 were most successful providing an overall pass rate of 87% and 99%, respectively (all protocol settings). In contrast, mAb clone 8G7G3/1 was less successful with a significantly lower pass rate of 20%. In particular, the increased use of the successful clones at the expense of 8G7G3/1 seemed to have a positive impact on the pass rate in this assessment. The RTU systems from Ventana (rmAb SP141) and Leica (mAb SPT24) was most successful, both providing a pass rate of 100% when used in compliance with vendor recommended protocol settings.  

MUM1: 259 laboratories participated and obtained a pass rate of 73% (46% optimal). The pass rate was highly influenced by the choice of the antibody (Ab). The well established clone MUMp1 was most successful both within a LD assay and as RTU system from Dako/Agilent. The Dako/Agilent RTU system provided an overall pass rate of 98% when used as “plug-and-play”. In contrast, a pass rate of 1% was observed for the mAb clone MRQ-43 being used by 13% of the participants.    

NKX3.1: 107 laboratories participated and obtained a pass rate of 82% (55% optimal). This was the second assessment of NKX3.1 in NordiQC and the pass rate increased from 65% seen in run 49, 2017. The rmAb clone EP356 was used by 66% of all participants and an optimal result could be obtained both within a LD assay and as a RTU system. Efficient HIER in an alkaline buffer and use of a sensitive and specific 3-step polymer / multimer based detection system gave the highest proportion of optimal results. The identification and use of testis as iCAPC for NKX3.1 seem to be supportive to monitor the level of analytical sensitivity as Sertoli cells express low-levels NKX3.1.    

SATB2: 105 laboratories participated and obtained a pass rate of 58% (33% optimal). This was the first assessment of SATB2 in NordiQC and focused on identification and characterization of colorectal carcinomas and neuroendocrine tumours. The relatively low pass rate was related to extended use of less successful Abs and not adequately calibrated IHC protocols. The rmAb clones EP281 and SP281 were most successful providing an optimal performance by the use of efficient HIER in an alkaline buffer and a 3-step detection system. Appendix, tonsil and testis were observed to be recommendable positive and negative tissue controls to monitor the level of analytical sensitivity and specificity.   

Breast module - run B29

Estrogen receptor (ER): 367 laboratories participated and obtained a pass rate of 94% (66% optimal). rmAb clones SP1 and EP1 and mAb clone 6F11 could all be used to provide an optimal result. 83% of the participants used RTU systems for the demonstration of ER. The RTU systems from Ventana, Dako and Leica used as “plug-and-play” assays provided an overall pass rate of 96%. In this assessment, a too weak or false negative staining reaction was the predominant feature of insufficient results. Tonsil was found to be recommendable as positive and negative tissue control.

Progesterone receptor (PR): 365 laboratories participated and obtained a pass rate of 90% (53% optimal). mAb clones 16, PgR 636, PgR 1294 and rmAb clone 1E2 could all be used to provide an optimal result. 79% of the participants used RTU systems for the demonstration of PR. The RTU systems from Ventana, Dako and Leica used as “plug-and-play” assays provided an overall pass rate of 98%. In this assessment, the insufficient results were mainly characterized by false positive results. This in particular being observed for off-label use of the Ventana RTU system based on rmAb 1E2.  Tonsil and uterine cervix in combination were found to be recommendable negative and positive tissue controls. 

HER2 IHC: 344 laboratories participated and obtained a pass rate of 93% (78% optimal). In this assessment, the FDA-/CE-IVD approved HER2 IHC assays HercepTest™ and PATHWAY® /4B5 were most the most accurate and successful assays providing a pass rate of 100% and 97%, respectively. LD assays were used by 25% and for this group a pass rate of 79% was obtained. The insufficient results primarily characterized by false negative results or a poor signal-to-noise ratio compromising the interpretation.   

HER2 ISH module - run H17

HER2 ISH: 133 laboratories participated in the technical assessment of HER2 (BR)ISH and obtained a pass rate of 75% (43% optimal). In this assessment, an increased pass rate especially compared to the latest run H16, but also to the recent runs, was seen. One of the contributing factors seems to be related to the expanded use of the VENTANA HER2 Dual ISH DNA Probe Cocktail, 800-6043 providing a pass rate of 80% on the expense of the less successful INFORM™ HER2 Dual ISH assay, 800-4422/780-4422 with a pass rate of 66%. In total 92% of the participants used one of these two, Ventana ISH systems. The insufficient results were mainly characterized by large negative areas and missing signals.

 

April 20th 2020

Søren Nielsen
Scheme director

NordiQC conference cancelled

It is with severe despondency and disappointment we hereby inform that we have decided to cancel the 4th NordiQC International conference planned 2-5th of June 2020. The global uncertainty and extensive travel restrictions related to covid-19 have had a tremendous impact on registrations for the conference, which has been status quo for more than 3 weeks now. The uncertainty has unfortunately and of central decisive importance for the cancellation also affected many of our speakers not being able to provide a definite confirmation of their presence. At present the incidence rate increases dramatically in Denmark and Europe and expected to continue with more restrictions to come concerning travel and meetings. We hereby have been put in a very risky situation to either postpone the cancellation of the conference to last minute due to no improvements for the covid-19 epidemic or to have a conference with a very reduced number of participants/speakers and hereby an abridged scientific meeting compared to the level expected from all stakeholders of our conference.

We hope that the conclusion to cancel the conference at this point is considered as due diligence, as we want to reduce the uncertainty and negative impacts related to the decision.

NordiQC reimburses all registration fees and sponsorship payments. If payment has been performed by bank account transfer, please send bank account data to us with registration number, IBAN and SWIFT codes.

Please receive our deepest apologies for all inconveniences.

Søren Nielsen

NordiQC director

Results - Run 57, B28, H16, C6 - ASMA Revision

Run 57, B28, H16 and C6 were accomplished from August to December 2019. In total, 450 laboratories participated in one or more of the modules offered. For the first time PD-L1 IHC assessment was segmented in PD-L1 KEYTRUDA® and PD-L1 TECENTRIQ® to align IHC test to treatment choice, cancer indications and scoring algorithms.

A short summary of the results is given below. Click on the epitope name to see the complete and detailed assessment report for the individual markers evaluated. In each assessment report, NordiQC intended to identify best practice methods and reliable and recommendable immunohistochemical critical assay performance controls (iCAPCs) to both guide the IHC protocol set-up and to monitor the IHC performance.

General module - run 57

ALK (lung): In total, 201 laboratories participated and obtained a pass rate of 84% (59% optimal). The mAb clone OTI1A4 and the rmAb clone D5F3 could both be used to provide an optimal staining result. To low concentration of the primary Ab and use of detection systems with low sensitivity were the main causes of insufficient staining reactions. The mAb clone ALK1, although useful when detecting translocations in lymphomas, cannot be used when detecting translocations in lung adenocarcinomas (and Merkel cell carcinomas) since it provides too low analytical sensitivity.

Bcl-2: In total, 319 laboratories participated and obtained a pass rate of 89% (64% optimal). Overall a very high pass rate and proportion of optimal results were observed. The well established clones as 124 and BCL2/100/D5 were most successful especially when applied within Ready-To-Use (RTU) systems e.g. from Dako/Agilent and Leica providing a pass rate of 100%. The rmAb clones E17 and EP36 unexpectedly showed an inferior performance and gave a negative reaction in one of the follicular lymphomas with t(14;18)(q32;q21) translocation.

Cytokeratin 8/18: In total, 238 laboratories participated and obtained a pass rate of 66% (41% optimal). The pass rate was highly influenced by the choice of the antibody (Ab). The clone cocktails against CK8/18 B22.1/B23.1 and EP17/30 and clone EP17 against CK8 were most successful both used within a laboratory developed (LD) assay or as RTU system. Abs towards CK18 showed an inferior performance in the included breast carcinoma due to partial loss of CK18 emphasizing the benefit to use Abs towards both CK8 and 18 for the identification of adenocarcinoma origin in tumour classification. 

MSH2: 258 laboratories participated and obtained a pass rate of 81% (53% optimal). The RTU systems for MSH2 from the two main vendors, Dako and Ventana both provided higher pass rates compared to LD assays. For optimal performance using these two RTU systems, protocols based on the vendor recommended settings were more successful compared to laboratory modified settings. This was seen e.g.for the Dako RTU system IR085 based on mAb clone FE11 with intended use for Autostainer Link 48. Using the RTU system according to the vendor recommended protocol, a pass rate of 100% was seen (86% optimal). If the RTU format IR085 was used by modified protocol settings, the pass rate was 87%, 56% optimal.    

SMAD4: 52 laboratories participated and obtained a pass rate of 42% (31% optimal). This was the first assessment of SMAD4 in NordiQC and focused on the use of IHC for identification of loss of SMAD4 expression caused by inactivation of the SMAD4 gene seen in e.g. pancreatic adenocarcinomas. The rmAb clone EP618Y was most successful and provided an optimal result on most IHC platforms, whereas the mAb clone B8 showed an inferior performance on the fully automated platforms from Dako (Omnis) and Ventana (BenchMark). Efficient HIER in an alkaline buffer and use of 3-step detection systems were main prerequisites for optimal demonstration of SMAD4.  

Breast module - run B28

Estrogen receptor (ER): 367 laboratories participated and obtained a pass rate of 93% (51% optimal). rmAb clones SP1 and EP1 and mAb clone 6F11 could all be used to provide an optimal result. The RTU systems from Ventana, Dako and Leica used as “plug-and-play” assays provided an impressive pass rate of 99%. In this assessment, a too weak or false negative staining reaction was the predominant feature of insufficient results, but also false positive results were observed.

HER2 IHC: 346 laboratories participated and obtained a pass rate of 86% (72% optimal). In this assessment, the FDA-/CE-IVD approved HER2 IHC assays HercepTest™ and PATHWAY® /4B5 were most successful and especially when used in concordance with the recommended protocol settings providing an overall pass rate of 90%. LD assays were used by 25% and for this group a pass rate of 65% was obtained. The insufficient results were characterized by both false negative and false positive results.   

HER2 ISH module - run H16

HER2 ISH: 137 laboratories participated and obtained a pass rate of 54% (39% optimal). Despite optimal protocol settings, a high proportion of technical insufficient results were seen, indicating that technical issues are influencing the quality and reproducibility of the BRISH assays. The most commonly used HER2 BRISH assay, INFORM™ HER2 Dual ISH 800-4422 / 780-4422 (Ventana/Roche), only provided a pass rate of 54% despite using appropriate, well-characterized protocol settings. The newly launched HER2 BRISH assay INFORM™ HER2 Dual ISH 800-6043 (Ventana/Roche) was more successful giving a pass rate of 84% when used by optimal protocol settings.

Companion module – C6

PD-L1 KEYTRUDA®: 182 laboratories participated and obtained a pass rate of 75% (46% optimal). This assessment for PD-L1 KEYTRUDA® primarily focused on the evaluation of the analytical accuracy of the IHC assays performed by the NordiQC participants to identify patients with NSCLCs and urothelial carcinomas to be treated with KEYTRUDA® as immune therapy. The CE IVD approved companion diagnostic assays as 22C3 SK006/GE006 Dako, 28-8 SK005 Dako and SP263 Ventana provided an overall pass rate of 89%, when used accordingly to package inserts. LD assays were widely applied by 60% of the participants. The overall pass rate for LD assays was 66%. The prevalent feature of insufficient results was characterized by false negative staining results changing the expected TPS or CPS status. 

PD-L1 TECENTRIQ®: 84 laboratories participated and obtained a pass rate of 76% (40% optimal). This assessment for PD-L1 TECENTRIQ® primarily focused on evaluation of the analytical accuracy of the PD-L1 IHC assays performed by the participating laboratories to identify patients with urothelial carcinomas or triple negative breast carcinomas (TNBC) to be treated with TECENTRIQ® as immune therapy. The PD-L1 SP142 companion diagnostic IHC assays from Ventana were most successful assays providing a pass rate of 94%. Other companion diagnostic assays and LD tests provided significantly lower pass rates of 67% and 65%, respectively. Insufficient results were typically characterized by a too strong staining reaction in tumour cells compromising the interpretation of immune cells.

ASMA

NordiQC has performed a revised assessment of ASMA run 55, 2019. In the original assessment, we observed a very low pass rate of only 17% and significantly inferior to the expected and obtained in previous runs. The assessment criteria were primarily focusing on the level of technical sensitivity and to a less extent on the diagnostic utility of IHC for ASMA. In the assessment a sufficient result required a positive staining in the GIST specimen and a negative staining reaction in this tumour thus lead to in an insufficient mark despite an otherwise optimal result obtained in all other tissues including the NordiQC recommended positive and negative tissue controls.   

The revised assessment of ASMA addressed both the technical level of sensitivity and primary diagnostic purpose of IHC for ASMA e.g. demonstration of ASMA in leiomyogene neoplasias. The pass rate in the revised assessment for ASMA was 58% and comparable to previous assessments for ASMA. However, only 9% were optimal and this still indicates an improved performance for ASMA should be required. 

The revised requirements and criteria for optimal performance were as applied in the original assessment but accepting a negative staining reaction of the GIST tumour as sufficient (good) provided that the IHC assays performed as expected (good or optimal) in all other tissue cores. 

Due to the low pass rate in run 55, many laboratories have asked for re-assessment of their IHC protocol in case of an insufficient result. These laboratories will receive a revised assessment mark for both the original submitted slide in run 55 and for the slide for re-assessment. 

Do not hesitate to contact NordiQC in need for clarifying questions.

At this point we also want to inform that we will repeat assessment for ASMA in run 59, 2020.

December 13th 2019
Søren Nielsen
Scheme director

Results - Run 56, C5

Run 56 (General module) and C5 (Companion module for PD-L1 NSCLC and Urothelial carcinoma) were accomplished from April to July 2019. In total 356 laboratories participated in one or both of the modules offered. For the first time, PD-L1 IHC testing also focused on Urothelial carcinoma in addition to NSCLC.  

A short summary of the results is given below. Click on the epitope name to see the complete and detailed assessment report for the individual markers evaluated. In each assessment report, NordiQC intended to identify best practice methods and reliable and recommendable immunohistochemical critical assay performance controls (iCAPCs) to both guide the IHC protocol set-up and to monitor the IHC performance.    

General module - run 56

C-MYC: 173 laboratories participated and obtained a pass rate of 57% (24% optimal). This was the first assessment of C-MYC in NordiQC and focused on the use of IHC for C-MYC in lymphoma classification. The rmAb clones EP121 and Y69 could both be used to provide optimal results. Efficient HIER in an alkaline buffer and use of 3-step detection systems were main prerequisites for optimal demonstration of C-MYC.  Laboratory developed (LD) assays based on one of these two Abs were more successful than corresponding Ready-To-Use (RTU) systems.   

CD117: 312 laboratories participated and obtained a pass rate of 87% (48% optimal). As seen in the previous assessments, the recently launched rmAbs EP10 and YR145 were most successful providing an overall pass rate of 97%, 78% optimal. The extended use of these Abs and implementation of best practice protocols for well established Abs such as the pAb 4502 have contributed to an increased pass rate compared to previous assessments.   

EpCAM: 256 laboratories participated and obtained a pass rate of 57% (34% optimal). Despite an increased pass rate compared to previous runs, the level was still low. For optimal performance, the selection of Ab must be made with focus on the IHC platform and reagents available in the laboratory. The most commonly used mAb clone Ber-EP4 require HIER in a special formulated buffer available for Dako IHC platforms and here provides high pass rates both within LD assays or as RTU system. On e.g. the Ventana BenchMark IHC platforms, mAb clones BS14 and VU-1D9 had significantly higher pass rates compared to mAb clone Ber-EP4.  

Melan A (MLA): 286 laboratories participated and obtained a pass rate of 65% (24% optimal). This assessment primarily addressed the performance in melanocytic lesions and not steroid hormone related lesions, only being relevant for mAb clone A103. mAb clone A103 was the most widely used Ab both within LD assays or RTU systems. For optimal performance, efficient HIER in an alkaline buffer and use of a sensitive and specific 3-step detection system were found to be essential. The RTU system IR/IS633 Dako based on mAb clone A103 for Autostainer Link 48 was most successful providing a pass rate of 100%, 58% optimal when using recommended protocol settings. The newly launched rmAb clone EP43 gave a promising performance and optimal results on all main IHC platforms.    

Mismatch repair protein 1 (MLH1): 259 laboratories participated and obtained a pass rate of 90% (64% optimal). A significantly increased pass rate was seen in this run compared to run 49, 2017 (59%). The RTU systems for MLH1 from the three main vendors, Dako, Leica and Ventana all provided higher pass rates compared to LD assays. For optimal performance using RTU systems, protocols based on the vendor recommended settings were more successful compared to laboratory modified settings. This was seen e.g.for the Dako RTU system IR/IS079 based on mAb clone ES05 with intended use for Autostainer Link 48. Using the RTU system according to the vendor recommended protocol, a pass rate of 100% was seen (93% optimal). If the RTU format IR/IS079 was migrated to Dako Omnis using same protocol settings, the pass rate was 77%, 27% optimal.   

Paired box gene-8 protein (PAX8): 264 laboratories participated and obtained a pass rate of 37% (12% optimal.) A surprisingly low pass rate was observed. 98% of the insufficient results were characterized by a generally too weak or false negative staining reaction in cells expected to be demonstrated. The combination of using challenging tissues with critical clinically relevant PAX8 expression levels and an extended use of Abs (e.g. clone MRQ-50) being less successful on the Ventana BenchMark and Dako Omnis platforms (used by the majority of all participants) had a negative impact on the pass rates.  

Companion module - run C5

PD-L1 (lung): 176 laboratories participated and obtained a pass rate of 80% (51% optimal). The assessment primarily focused on the accuracy of the PD-L1 IHC assays for TPS status in NSCLCs to guide treatment with KEYTRUDA®. The CE IVD approved companion diagnostic assays as 22C3 SK006/GE006 Dako and SP263 Ventana provided a pass rate of 92% and 91%, respectively, when used accordingly to package inserts. LD assays were widely applied by 63% of the participants. The overall pass rate for LD assays was 74%. The prevalent feature of insufficient results was characterized by false negative staining results changing the TPS status from high to low or low to negative.  

PD-L1 (uro): 93 laboratories participated and obtained a pass rate of 76% (50% optimal). The assessment focused on the accuracy of the PD-L1 IHC assays for CPS status in Urothelial carcinomas to guide treatment with KEYTRUDA®. Similar to PD-L1 IHC for NSCLC, companion diagnostic assays provided a superior pass rate compared to LD assays. LD assays was used by 66% of the participants and an overall pass rate of 70% was seen. This was in contrast to the Dako pharmDx 22C3 PD-L1 SK006 that provided a pass rate of 91%. 

 

July 9th 2019
Søren Nielsen
Scheme director

 

Segmentation of Companion PD-L1 module focusing on intended use of PD-L1 IHC

PD-L1 IHC testing has a significant impact on patient eligibility for tailored specific immune oncology and require diagnostic accuracy. This also underlines the need of appropriate EQA methods to evaluate the performance of PD-L1 IHC testing among the participating laboratories. 

At present, PD-L1 IHC testing is based on the alignment of treatment choice / cancer indication / IHC test method including scoring algorithms. This means that is mandatory to focus on the purpose of a PD-L1 test to evaluate the diagnostic accuracy and adequacy. Based on this NordiQC intends, from run C6 and onwards, to offer 2 segments in the companion module for PD-L1 IHC testing;

  • One segment focusing on PD-L1 IHC in NSCLC and Urothelial carcinoma for treatment with KEYTRUDA®. Laboratories must score the stained slides using the associated TPS- and CPS scoring systems.
  • One segment focusing on PD-L1 IHC in triple negative breast cancer and Urothelial carcinoma for treatment with TECENTRIQ®. Laboratories must score the stained slides using the associated IC-scoring system. 

The diagnostic accuracy will be evaluated on the participants performance compared to “gold standard / reference standard” methods and results characterized by the respective associated and approved companion diagnostic PD-L1 IHC assays and relevant scoring methods / cut-offs. The participants can use both laboratory developed and companion diagnostic assays for PD-L1 but need to specify for which purpose. The modification and updated segmentation of PD-L1 IHC testing will enable NordiQC to give more valuable information to the participants on their diagnostic accuracy. There are many additional purposes including different treatment options for PD-L1. At this point NordiQC has decided to focus on the most widely tested areas in PD-L1 IHC testing but will update the segmentation accordingly to any changes being required.

Søren Nielsen
Scheme director

Results - Run 55, B27, H15

Run 55 (General module), B27 (Breast cancer IHC module), and H15 (HER2 ISH module) were accomplished December 2018 – April 2019. A very short summary of the results is given below. Click on the epitope name to see the complete general assessment results for each marker, including major causes of insufficient results, immunohistochemical critical assay performance controls (iCAPCs), and recommendable clones and protocols. Individual results are available after login on the homepage.

General module - run 55

Alpha-smooth mucle antigen (ASMA): 281 labs participated, 17% sufficient, 9% optimal based on a very challenging test material. mAb clone BS66 as concentrate gave optimal results in all of 7 cases. The most used mAb clone 1A4 could only give optimal results with the Dako Autostainer and Leica Bond platforms. rmAb clone EP188 could give optimal results on the Ventana platform with a combined proteolytic and heat induced pretreatment. 

Bcl-6: 279 labs participated, 77% sufficient, 45% optimal. The mAb clones PG-B6p and LN22 provided the highest proportion of sufficient and optimal results applied as an RTU format (Dako and Leica, respectively). The RTU system based on the mAb clone GI191E/A8 (Ventana) provided a lower pass rate and was challenged by poor signal-to-noise ratio and false positive results.

Cytokeratin 5 (CK5): 263 labs participated, 44% sufficient, 25% optimal. Optimal staining results could be obtained with the mAb clone XM26, and the rmAbs clones BSR55, EP24and SP27used as concentrates. However, SP27 has in NordiQC studies (unpublished) shown positive reaction in lung adenocarcinomas being negative for other CK5 antibodies as well as p40. The significance of this is currently uncertain. The ascites produced mAb clone D5/16 B4 was less successful due to low analytical sensitivity in combination with the risk of false positive “Mouse Ascites Golgi” staining reaction.

SOX10: 204 labs participated, 89% sufficient, 68% optimal. The mAb clones BC34, BS7, ZM10, and the rmAb clones EP268 and SP267 could all be used to obtain optimal results for SOX10 with appropriate protocols. The RTU system 760-4968 (Ventana) based on the rmAb clone SP267, showed superior performance. 

Wilms’ tumour-1 protein (WT1): 291 labs participated, 90% sufficient, 60% optimal. The mAb clones 6F-H2, WT49 and the rmAb clones EP122 and D8I7F are all recommendable antibodies with appropriate protocols. A combined HIER–protease retrieval protocol provided a significant improvement in the signal-to-noise ratio for the mAb clone 6F-H2 on the Ventana platform (both as concentrate and RTU system).

Breast cancer IHC module - run B27

Estrogen receptor (ER): 348 labs participated, 90% sufficient, 54% optimal. rmAb clones SP1 and EP1 and mAb clone 6F11 could all be used to provide an optimal result. The corresponding Ready-To-Use (RTU) system from Ventana/Roche (rmAb SP1) provided the highest proportion of sufficient and optimal results.

HER2 IHC: 324 labs participated, 86% sufficient, 83% optimal. The laboratory modified protocols generally gave better results than vendor recommended protocols for the Ventana PATHWAY® and Dako HercepTest™. However, modifications must be meticulously validated by the end-users on a large cohort of breast carcinomas (see ASCO/CAP 2013 guidelines).

HER2 ISH module - run H15

HER2 ISH: 122 labs participated, 72% sufficient, 50% optimal. Despite optimal protocol settings, a high proportion of technical insufficient results were seen, indicating that technical issues are influencing the quality of the BRISH assays. Thus, the most commonly used HER2 BRISH assay, INFORM™ HER2 Dual ISH (Ventana/Roche), only provided a pass rate of 71% despite using appropriate, well-characterized protocol settings.

 

April 17th 2019

 

Mogens Vyberg

Scheme director

 

Results - Run 54, B26, H14, C4

Run 54 (General module), B26 (Breast cancer IHC module), H14 (HER2 ISH module) and C4 (Companion module) was accomplished September to December 2018. A very short summary of the results is given below. Click on the epitope name to see the complete general assessment results for each marker, including major causes of insufficient results, immunohistochemical critical assay performance controls (iCAPCs), and recommendable clones and protocols. Individual results are available after login on the homepage.

General module - run 54

Carcinoembryonic antigen (CEA): 272 labs participated, 63% sufficient, 37% optimal. The mAb clones CEA31 and COL-1 give high performance irrespective of stainer platform. While the mAb clone II-7 showed significantly lower analytical sensitivity. Heat induced epitope retrieval in an alkaline buffer, careful calibration of the primary antibody anduse of sensitive 3-step multimer/polymer detection systems were generally important prerequisites for an optimal staining result.The mAb clone TF3H8-1 (Ventana, 760-2507) cross reacts with other protein and should not be used.

CD8: 262 labs participated, 65% sufficient, 52% optimal. Many mouse monoclonal antibodies may give optimal results, provided an optimally calibrated protocol.  In contrast,the majority of assays based on the rmAb clones SP16 and SP57, both as concentrated formats or RTU systems, were challenged by false positive staining reaction in epithelial cells hindering interpretation of the specific signal for CD8.

GATA3: 245 labs participated, 76% sufficient, 50% optimal. Used within a LD-assay, the performance of the mAb L50-823 was significantly influenced by the company/distributor of the primary Ab). Efficient HIER, preferable in an alkaline buffer, careful calibration of the primary antibody concentration and use of a sensitive detection system were the most important prerequisites for optimal staining results.

Pan Cytokeratin (CK-PAN): 296 labs participated, 62% sufficient, 38% optimal. The mAb clone cocktails AE1/AE3, alone or combined with 5D3 or PCK26, and mAb clone BS5 can all be recommended but the epitope retrieval method must be specifically tailored to the clone/cocktail applied. Applying the AE1/AE3 on the Bond platform (Leica) is challenging, and the mAb clone MNF116 should not be used due to poor performance. 

Podoplanin (Podop): 225 labs participated, 64% sufficient, 36% optimal. The concentrated format of the mAb clone D2-40 could provide optimal staining results on all four main stainer platforms – Dako Autostainer, Dako Omnis, Leica Bond and Ventana BenchMark, if the protocols were optimized. In general, the performance of the Ready-To-Use systems was inferior to the laboratory developed assays The Ventana 760-4395 RTU showed low analytical sensitivity whereas the DAKO IR/IS072 RTU was troubled by false positive staining reactions on the Dako Autostainer. 

Breast cancer IHC module - run B26

Estrogen receptor (ER): 355 labs participated, 70% sufficient, 31% optimal, which are markedly lower scores than in the previous four runs, probably due to more challenging tumours in the TMA. rmAb clone SP1 is a highly robust Ab performing better than rmAb clone EP1 and mAb clone 6F11.       

HER2 IHC: 335 labs participated, 76% sufficient, 71% optimal. This is a decline compared to the previous run, probably due to more challenging tumours in the TMA. While the Ventana Pathway is very robust, about half of the laboratories using the Dako pAb A0485 either in the Herceptest or in a laboratory developed test, got an insufficicent resultat. Interestingly, all labs using mAb clone CB11 in a laboratory developed test with an optimal protocol got sufficient results, while those using the Oracle™ mAb clone CB11 assay, all got insufficient results.

Progesterone receptor (PR): 349 las participated, 85% sufficient, 66% optimal. The most widely used Ab clones 16, PgR 636, PgR 1294 and 1E2 could all be used to obtain an optimal result. Irrespective of the clone applied, efficient HIER and careful calibration of the primary antibody were mandatory for optimal performance. Ready-To-Use (RTU) systems and laboratory developed tetst showed a similar performance.

HER2 ISH module - run H13

HER2 ISH: 193 participated either by sending 127 labs participated, 71% sufficient, 41% optimal. Optimal demonstration and evaluation of the HER2 gene amplification status in all five cores of the multi-tissue block could be obtained by all the applied dual-colour systems and the Ventana/Roche HER2 SISH one colour system. Artefacts as silver precipitates, excessive background staining or negative areas were most likely caused by technical issues as slides drying out during the staining process or inadequate washing. Laboratories performing FISH achieved a significantly higher consensus rate for the interpretation of HER2 amplification status compared to laboratories performing BRISH. 

Companion module - run C4

PD-L1 (lung): 163 laboratories participated, 86% sufficient, 57% optimal. Both the companion diagnostic PD-L1 IHC assay 22C3, SK006 Dako/Agilent, and the complementary diagnostics 28-8, SK005 Dako/Agilent, and SP263, 790-4905 Ventana/Roche as well as the LD assays for PD-L1 provided a high pass rate. 

December 14th 2018


Mogens Vyberg

Scheme director

Results - general module run 53

B-cell specific activator protein (BSAP, PAX5): 198 labs participated, 86% sufficient, 46% optimal. Many clones could give optimal results, best performance was seen with DAK-Pax5 (Dako/Agilent). In contrast, rmAb clone 34 (Ventana/Roche, Cell Marque) gave a very low proportion of optimal results due to a generally poor signal-to-noise ratio.

Chromogranin (CGA): 296 labs participated, 76% sufficient, 51% optimal. mAb clone LK2H10 gave superior performance, both as Conc (many companies) and RTU systems (760-2519, Ventana/Roche, and others). In contrast, mAb clone DAK-A3 as Conc (Dako/Agilent), and mAb clone 5H7, both as Conc (Leica) and RTU (PA0430/PA0515, Leica), gave unsatisfactory results.

E-Cadherin (ECAD): 298 labs participated, 89% sufficient, 65% optimal. The RTU assays based on mAb clone NCH-38 (IS/IR/GA059, Dako/Agilent) and mAb clone 36 (790-4497, Ventana/Roche) gave a high proportion of optimal results. In contrast, RTU assays based on rmAb clone EP700Y (760-4440, Ventana/Roche, and 246R-18, Cell Marque) revealed poor signal-to-noise ratios or false positive staining reactions hampering interpretation of the specific signal for ECAD.

Mismatch repair protein PMS2 (PMS2): 246 labs participated, 89% sufficient, 58% optimal. The highest proportion of sufficient staining results was seen with rmAb clone EP51 based Autostainer RTU system (Dako/Agilent) and the rmAb clone EPR3947 based Benchmark RTU system (Ventana/Roche). However, as for the latter it is noteworthy that 80 out of 83 labs (96%) modified the protocol significantly in order obtain a better staining reaction, in fact changing the RTU system to a lab developed system.

Octamer transcription factor-3/4 (OCT3/4): 189 labs participated, 94% sufficient, 83% optimal. The Conc format of the mAbs clones MRQ-10 and N1NK provided a high proportion of optimal staining results on all the four main stainer platforms: Omnis (Dako/Agilent), Autostainer (Dako/Agilent), Bond (Leica) and BenchMark (Ventana/Roche). Also the mAb clone C-10 could provide optimal staining results on all four main stainer platforms, but the proportion of optimals were significantly lower on the BenchMark and Bond compared to the Omnis and Autostainer platforms.

Reassesment for run 53 will open together with run 54, B26, H14, C4 on 1st August.

July 2018
Mogens Vyberg
NordiQC director

Results - run 52, B25, H13 & C3

Run 52 (general module), B25 (breast cancer IHC module), H13 (HER2 ISH module) and C3 (Companion module) was accomplished December 2017 to April 2018. A very short summary of the results is given below. Click on the epitope name to see the complete general assessment results for each marker, including major causes of insufficient results, immunohistochemical critical assay performance controls (iCAPCs), and recommendable clones and protocols. Individual results are available after login on the homepage.

General module - run 52

Calretinin (CR): 269 labs participated, 72% sufficient, 45% optimal. Many clones may give optimal results.  Ready-to-Use (RTU) assays generally performed better than lab developed (LD) assays indicating a need for better calibration of the latter. RTU assays should only be used on the platform for which they are developed!

MSH6: 242 labs participated, 52% sufficient, 33% optimal. Rabbit monoclonal antibodies (rmAbs) generally performed well, while the mouse monoclonal antibodies (mAbs), clone 44 and PU29 consistently failed.

Synaptophysin (SYP): 308 labs participated, 75% sufficient, 33% optimal. The lowest proportion of acceptable results both as RTU and LD assays were seen with mAbs clone SY38 (1 out of 3 sufficient) and clone MRQ-40 (9 out of 61 optimal).

Terminal deoxynucleotidyl Transferase (TdT): 225 labs participated, 82% sufficient, 39% optimal. The mAb clone SEN28 and the rmAb clone EP266 were the most successful antibodies for TdT. Efficient HIER, preferable in an alkaline buffer in combination with a sensitive and specific 3-step polymer/multimer based detection system gave the highest proportion of optimal results. Polyclonal antibodies gave virtually no optimal results.

Vimentin (VIM): 308 labs participated, 74% sufficient, 43% optimal. The mAb clones V9 and 3B4 and rmAb clone SP20, HIER in an alkaline buffer, careful calibration of the primary Ab and application of a sensitive 3-step polymer/multimer based detection system were the most important parameters for an optimal performance. The clone V9 RTU assay from Roche/Ventana (790-2917) performed less well than V9 RTU assays from other companies.

Breast cancer IHC module - run B25

Estrogen receptor (ER): 361 labs participated, 92% sufficient, 57% optimal. Both LD and RTU assays generally perform well with the mAb clone 6F11 and rmAb clones SP1 and EP1. However, as for the Leica Bond mAb 6F11 RTU (PA009/PA0151) the lab modified protocols gave better results than the vendor recommended protocols.

HER2 IHC: 342 labs participated, 96% sufficient, 81% optimal. Despite an overall improvement of LD HER2 assays from run B1 to B25, the pass rate and proportion of optimal results is still inferior to the results of FDA/CE-IVD approved systems as PATHWAY® /CONFIRM™ and HercepTest™.

HER2 ISH module - run H13

HER2 ISH: 127 labs participated, 71% sufficient, 43% optimal. Optimal demonstration and evaluation of the HER2 gene amplification status could be obtained by all the applied dual-colour and one colour systems. For the most commonly used HER2 BRISH assay, the INFORM™ HER2 Dual ISH (Ventana/Roche), a technically adequate result was provided in 65% of the slides using appropriate and vendor recommended protocol settings. A similar issue has been observed consistently in the latest NordiQC HER2 BRISH assessments, indicating a challenge for the present assay to provide a reproducible performance. As this test is used by 79% of all participating laboratories applied with appropriate protocol settings, it significantly affects the pass rate. At present, no recommendations on how to improve the end result have been identified.

Companion module - run C3

PD-L1: 146 laboratories participated, a pass rate of 91% was observed. Both the three companion diagnostic PD-L1 IHC assays 28-8, SK005 Dako/Agilent, 22C3, SK006 Dako/Agilent and SP263, 790-4905 Ventana/Roche and the LD assays for PD-L1 provided a high pass rate. However, the assessment of C3 was challenged by less than optimal material circulated. It should be emphasized that the LD assays must be carefully validated before use for diagnostics.

April 20th 2018
Mogens Vyberg
Scheme director

Results - module 51, B24, H12, C2

Run 51 (general module), B24 (breast cancer IHC module), H12 (HER2 ISH module) and C2 (Companion module) was accomplished August to December 2017. A very short summary of the results is given below. Click on the epitope name to see the complete general assessment results for each marker, including major causes of insufficient results, immunohistochemical critical assay performance controls (iCAPCs), and recommendable clones and protocols. Individual results are available after login on the homepage.

General module 51

lu-ALK (lung anaplastic lymphoma kinase): 189 labs assessed, 61% sufficient, 38% optimal. The mAb clone OTI1A4 and the rmAb clone D5F3 were the most successful Abs for demonstration of EML4-ALK translocation in lung adenocarcinoma. For mAb clone 5A4 the analytical sensitivity was significantly lower. mAb ALK1 must not be used for lung adenocarcinoma. The Ventana clone D5F3 RTU system was the most successful assay with an overall pass rate of 100% when not modified by the laboratories. HIER at high pH and use of a sensitive 3-step polymer/multimer based detection system are crucial for an optimal performance.

AMACR (α-methylacyl-CoA racemase): 250 labs assessed, 93% sufficient, 55% optimal. The rmAb clones 13H4 and SP116, mAb clone EPMU1, and pAb p504S/CP200 were all robust Abs for demonstration of AMACR. Efficient HIER in an alkaline buffer and careful calibration of the primary Ab, in combination with a sensitive IHC system (3-step polymer/multimer system), were the main prerequisites for an optimal staining result.

CD30: 282 labs assessed, 83% sufficient, 50% optimal. The mAb clones Ber-H2, CON6D/5 and JCM182 could all be used to obtain optimal staining results. Efficient HIER in an alkaline buffer or modified low pH buffer, in combination with a sensitive and specific IHC system were the main prerequisites for optimal performance.

CD117: 277 labs assessed, 63% sufficient, 25% optimal. The rmAb clones YR145 and EP10 were the most successful both in a laboratory develop (LD) assay and in an RTU format, as the overall pass rate was 94% and the proportion of optimal results was 73%. Efficient HIER in an alkaline buffer, careful calibration of the primary Ab and use of a 3-step polymer/multimer detection system provided the highest proportion of optimal results. Assays based on the pAb A4502 (Dako) were used by the majority of participants, but only 53% were sufficient and 6% optimal. A similar pattern was seen with the RTU system based on the rmAb 9.7 (Ventana), where only 53% were sufficient and 3% optimal. Both Abs are difficult to calibrate to an acceptable level.

PAX8: 213 labs assessed, 56% sufficient, 20% optimal. Optimal staining results could be obtained with the mAbs clones BC12 and PAX8/1492, the pAb 10336-1-AP, 363A and CP379 and particularly the recently introduced rmAb clones EP298, SP348 and ZR-1. The most widely used PAX8 antibody, mAb clone MRQ-50, performed satisfactory on both the Leica Bond and Dako Autostainer platforms but had a very low pass rate on the Ventana BenchMark platform, both in a laboratory develop (LD) assay and in an RTU assay. The highest proportion of optimal results was obtained performing HIER in an alkaline buffer for at least 20 min. (32 min. for Ventana BenchMark) and use of a sensitive and specific 3-step polymer/multimer based detection system.

Breast cancer IHC module - run B24

ER: 386 labs participated, 92% sufficient, 71% optimal. Both lab developed and RTU assays generally perform well with the mAb clone 6F11 and rmAb clones SP1 and EP1. However, as for the RTUs the lab modified protocols generally gave better results than the vendor recommended protocols from Dako/Agilent, Leica and VMS/Roche: Grouped together, 163 out 203 (80%) of the lab modified RTU protocols gave optimal results, compared to 50 out of 74 (68%) of vendor recommended protocols. It is noteworthy that 203 out of 277 labs (73%) found need to modify the vendor recommended protocols in order to optimize the staining reaction.

HER2 IHC: 369 labs participated, 97% sufficient, 78% optimal. Despite an overall improvement of the pass rate for lab developed HER2 assays from run B1 to B24, the pass rate and proportion of optimal results is still inferior to the FDA/CE-IVD approved systems as PATHWAY® /CONFIRM™ and HercepTest™. In general, the three FDA-/CE-IVD approved HER2 assays provided a proportion of optimal results of 85% (226 of 266), whereas only 59% (61 of 103) of LD HER2 assays were assessed as optimal. On the other hand laboratory modified RTU assays seemed to perform better than the vendor recommended: Compiling the resuls of Agilent/Dako and Roche/VMS assays, 90 of 120 (75%) stains were optimal with the vendor recommended protocols, while 119 of 131 (91%) with the laboratory modify protocols. It is noteworthy that 146 out of 266 labs (55%) found need to modify the vendor recommended protocols in order to optimize the staining reaction.

PR: 385 labs participated, 99% sufficient, 77% optimal. Both lab developed assays and RTU assays generally perform well with the most widely used Abs clones 16, PgR 636, PgR 1294 and 1E2.

Breast cancer ISH module - run H12

HER2 ISH: 115 labs participated, 64% sufficient, 37% optimal. Technically optimal demonstration of HER2 BRISH could be obtained by the commercially available two-colour HER2 systems INFORM™ HER-2 Dual ISH (Ventana) and ZytoDot® 2C (ZytoVision), and the single-colour HER2 systems INFORM™ SISH system (Ventana) and ZytoDot® (ZytoVision). For all systems, retrieval settings – HIER and proteolysis – must be carefully balanced to provide sufficient demonstration of signals and preserve morphology. Despite optimal protocol settings, a high proportion of technically insufficient results were seen, which currently cannot be explained.

Companion module - run C2

PD-L1: 145 laboratories participated, a pass rate of 84% was observed. The three companion diagnostic PD-L1 IHC assays 28-8, SK005 Dako/Agilent, 22C3, SK006 Dako/Agilent and SP263, 790-4905 Ventana/Roche were most successful providing a pass rate of 95%. Laboratory developed (LD) assays for PD-L1 provided a pass rate of 73%, (in comparison, a pass rate of 20% in C1 for this group was observed). Access to best practice protocol settings for LD assays are being developed and published supporting the implementation of LD assays. However, it must be emphasized that the LD assays must be carefully validated before use for diagnostics.


December 15th 2017
Mogens Vyberg
Scheme director

QuIP/NordiQC Workshop Brugge 13-15 June '18

This 1st QuIP/NordiQC Workshop in Brugge, Belgium is aimed to give an update on basic and advanced immunohistochemical methods applied in diagnostic pathology, particularly for the classification of the unknown primary tumour as well as breast, lung and haematolymphoid tumours. The workshop will particularly focus on
- understanding the three phases in the technical test approach
- planning and diagnostic use of antibody panels
- optimization of staining protocols 
- use of recommended tissue controls 
- identification and handling of technical and diagnostic pitfalls
- image analysis of immunostains for diagnostic use

The target group comprises biomedical technicians and scientists or equivalent with some experience in immunohistochemistry, and residents and consultants in clinical and surgical pathology with special interest in the technical parts of immunohistochemistry. Maximum number of participants: 70. 

Programme and practical details

Click this link for registration NB! Fully booked, you may register for the waiting list

Results - module 50 available

Run 50 was accomplished January to July 2017. A very short summary of the results is given below. Click on the epitope name to see the complete general assessment results for each marker, including immunohistochemical critical assay performance controls (iCAPCs), recommendable clones and protocols, as well as major causes of insufficient results.
Individual results are sent to all participants by email.

Results general module - run 50

CD23: 270 labs assessed, 89% sufficient, 55% optimal. All stains deemed insufficient showed too weak reactions. Many clones provided optimal results. Irrespective of the clone, HIER in an alkaline buffer and careful calibration of the primary Ab in combination with a sensitive 3-step detection system are critical parameters for a sufficient result. On the Ventana BenchMark platform, rmAb SP23 should be preferred rather than mAb 1B12.     RTU systems for CD23 from the main IHC system providers were most successful.

CK19: 245 labs assessed, 82% sufficient, 42% optimal. HIER in an alkaline buffer, and careful calibration of the primary Ab in combination with a sensitive 3-step detection system are critical parameters for a sufficient result. Proteolytic pretreatment as single retrieval method is incompatible with an optimal result.

ERG: 130 labs assessed, 67% sufficient, 38% optimal. The prevalent feature of insufficient results was too weak or false negative staining reaction. HIER in an alkaline buffer, and careful calibration of the primary Ab in combination with a sensitive 3-step detection system are critical parameters for a sufficient result. Primary Ab selection is also important: mAb 9FY was less successful compared to rmAb clone EP111 and EPR3864. The Agilent/Dako RTU system based on rmAb clone EP111 provided the highest pass rate and proportion of optimal results.

MSH2: 231 labs assessed, 79% sufficient, 50% optimal. Most stains deemed insufficient showed too weak reactions but cases with false positive reactions or poor signal-to-noise ratio were also seen. The performance of mAb clone 25D12 was, as seen in previous asessments, inferior to mAb clones FE11 and G219-1129. mAb clone FE11 performs less well on the Ventana BenchMark platform, while with mAb G219-1129 no optimal results were seen with the Dako Autostainer and Leica Bond platforms. RTU systems for MSH2 from Agilent/Dako and Roche/Ventana were most successful.

S100: 299 labs assessed, 82% sufficient, 23% optimal. pAbs against S100 were most successful. HIER in an alkaline buffer and a careful calibration of the primary Ab are critical parameters for a sufficient result. Omission of HIER or use of proteolytic pre-treatment cannot be recommended. RTU systems based on pAbs from the main IHC providers gave relatively high pass rates, but few optimal results.

SMH: 109 labs assessed, 78% sufficient, 42% optimal. HIER in an alkaline buffer, and careful calibration of the primary Ab in combination with a sensitive 3-step detection system are critical parameters for a sufficient result. mAb clone SMMS-1 was the most widely used primary Ab and provided optimal results on all main IHC systems both within laboratory developed assays and RTU systems.

July 10th 2017
Mogens Vyberg, Scheme director

Results - module 49, B23, H11 & C1 available

Login to see individual results

CD5: 278 labs assessed, 92% sufficient, 68% optimal. Ready-To-Use (RTU) systems obtained superior pass rates and higher proportion of optimal results compared to laboratory developed (LD) tests. Within LD tests, optimal results required efficient HIER, preferable in alkaline buffer, and careful calibration of the primary Ab titre.

CK-LMW: 229 labs assessed, 80% sufficient, 31% optimal. The mAb clone cocktail B22.1/B23.1, rmAb clone cocktail EP17/30 and rmAb clone EP17 were the most successful antibodies for immunohistochemical demonstration of CK-LMW. In contrast mAb clone 34βH11 and in particular clones CAM5.2 are less successful and prevent improvement of the overall pass rate for CK-LMW.

MLA: 255 labs assessed, 60% sufficient, 32% optimal. The Ready-To-Use system for MLA from Dako (IR633/IS633 – AS48) based on mAb clone A103 applied according to the vendor recommended protocol settings provided the highest proportion of sufficient and optimal results. However, the clone showed less successful performance on the fully automated IHC platforms Omnis (Dako) and BenchMark (Ventana).

MLH1: 224 labs assessed, 59% sufficient, 30% optimal. Clone ES05 was most successful, both as concentrate and as RTU format (Dako/Agilent and Leica/Novocastra). The mAb clone M1 could not be used to obtain optimal staining for MHL1 due to aberrant nuclear staining in colon adenocarcinoma known to lack MLH1 expression.

NKX3.1: 49 labs assessed, 65% sufficient, 45% optimal. rmAb clone EP356 and pAb CP422 were the most successful antibodies. With LD assays, efficient HIER in an alkaline buffer and use of a sensitive and specific 3-step polymer/multimer based detection system gave the highest proportion of optimal results. Testis and normal prostate can be used as positive tissue controls

PSA: 284 labs assessed, 89% sufficient, 68% optimal. Careful calibration of the Ab concentration and sufficient HIER are required.

Results breast cancer IHC module - run B23

ER: 394 labs assessed, 92% sufficient, 56% optimal. The most used clones 6F11, EP1 and SP1 are all recommended.     RTU systems obtained superior pass rates and higher proportion of optimal results compared to LD assays. For LD assays, too low concentration of the Ab and insufficient HIER were major reasons for insufficient assays.

HER-2 IHC: 372 labs assessed, 95% sufficient, 82% optimal. The FDA-/CE-IVD approved PATHWAY®/CONFIRM™ (Ventana) and HercepTest™ (Dako) were the most precise assays for the semi-quantitative IHC determination of HER-2 protein expression. LD assays produced a lower pass-rate and were less precise for the HER-2 status requiring an additional ISH test for final evaluation. Inclusion of 2+ tumours with and without HER-2 gene amplification in the control material for both EQA and internal quality control is essential to evaluate precision and performance stability of the IHC HER-2 assays used by laboratories.

Results companion module - run C1

PD-L1: 68 labs assessed, 50% sufficient, 37% optimal. Suboptimal results are mostly related to unexplained technical issues, which prevent recommendations for improvement. The Dako/Agilent 22C3 pharmDx assay SK006, applied by protocol settings in compliance with the vendor recommendations, was most successful with an overall pass rate of 92%. LD assays were used by 44% of the labs and a pass rate of 20% was seen.

Results breast cancer ISH module - run H11

HER-2 ISH: 123 labs assessed (BRISH only), 60% sufficient, 32% optimal. Suboptimal results are mostly related to unexplained technical issues, which prevent recommendations for improvement. 

 

April 21st 2017

Mogens Vyberg

 

Scheme director 

Welcome to Aalborg
Results - module 48, B22, H10 available

Runs 48/B22/H10 were accomplished from September to December 2016. A very short summary of the technical test results is given below. Click on the epitope name to see the complete general assessment results for each marker, including recommended clones and protocols, as well as major causes of insufficient staining results, and for HER2 IHC/ISH and Ki67 also concordance of interpretation. Individual results are available for the participants by logging in.

Results general module - run 48

CDX2: 268 labs assessed, 80% sufficient, 58% optimal. Careful calibration of the Ab concentration and a sensitive visualization system are required. The old clones CDX-88 and AMT28 cannot be recommended. DAK-CDX2 should not be applied on the Ventana BenchMark platform.

Desmin (DES): 275 labs assessed, 87% sufficient, 48% optimal. Efficient Heat induced epitope retrieval (HIER) and a sensitive detection system are mandatory for an optimal result. Clone D33 should not be used on the Dako Omnis platform. RTU VMS mAb DE-R-11 (760-2513 for Ultra/XT) performed better with laboratory modified protocols than with the vendor recommended protocol (61% vs 0% optimal). In contrast RTU Dako mAb D33 (IR606/IS606 for AS48) performed better with the vendor recommended protocol than with laboratory modified protocols (80% vs 57% optimal).

MUM1: 211 labs assessed, 60% sufficient, 40% optimal. Efficient HIER, careful calibration of the Ab concentration and a sensitive visualization system are required for optimal performance. The clones rmAb MRQ-43, mAb MRQ-8 and mAb BC5 all produce insufficient results and cannot be recommended.

p40: 188 labs assessed, 74% sufficient, 42% optimal. Careful calibration of the Ab concentration and a sensitive visualization system are required. Polyclonal Abs cannot be recommended. RTU VMS mAb BC28 (790-4950 for Ultra/XT) performed better with laboratory modified protocols than with the vendor recommended protocol (57% vs 40% optimal).

p63: 274 labs assessed, 82% sufficient, 44% optimal. Efficient HIER, careful calibration of the Ab concentration and a sensitive visualization system are required for optimal performance. Clone 7JUL cannot be recommended, as almost all labs failed with this clone. RTU VMS mAb 4A4 (790-4509 for Ultra/XT) performed better with laboratory modified protocols than with the vendor recommended protocol (60% vs 20% optimal).

SOX10: 121 labs assessed, 68% sufficient, 50% optimal. Careful calibration of the Ab concentration and a sensitive visualization system are required. Polyclonal Abs cannot be recommended.

Results breast cancer IHC module - run B22

HER2 IHC: 387 labs assessed, 84% sufficient, 71% optimal. Pathway®/ConfirmTM (Ventana) and HercepTestTM (Dako) gave pass rates above 90% while the pass rates for OracleTM (Leica) and the laboratory developed kits were too low. Several labs are using approved systems off-label, which gives a lower pass rate and should be omitted, also for legal reasons. 

Ki67: 409 labs assessed, 93% sufficient, 69% optimal. Efficient HIER and careful calibration of the primary Ab concentration are mandatory. RTU systems gave a higher pass rates than laboratory developed protocols. MIB-1 showed an inferior performance on Bond, Leica.

Results breast cancer ISH module - run H10

HER2 ISH: 118 labs assessed (BRISH only), 68% sufficient, 32% optimal. Suboptimal results are mostly related to unexplained technical issues, which prevent recommendations for improvement. 

 

December 9th 2016

Mogens Vyberg

Scheme director 

 

Results general module - run 47

Runs 47 was accomplished April to July 2016. A very short summary of the tests is given below. Click on the epitope name to see the complete general assessment results for each marker, including recommended clones and protocols, and major causes of insufficient staining results. Individual results will be sent to participant by email.

Figure: Serial sections of GIST stained for CD117 in two labs. Left: optimal, right: false negative due to an insufficient protocol.

CD117: 272 participants, 47% sufficient, 14% optimal. Best results were obtained with rmAb clones YR145 and EP10, both as concentrates and in RTU formats, while pAb 4502 (Dako) gave less satisfactory results, and rmAb clone 9.7 (RTU 790-2951, Ventana) performed poorly. HIER in an alkaline buffer in combination with careful calibration of the primary Ab were the most critical parameters for a sufficient result. See photo above

CEA: 255 participants, 42% sufficient, 17% optimal. Best results were obtained with mAb clones CEA31 (both as concentrates and in RTU formats) and COL-1 (as concentrate). mAb clone II-7 gave less satisfactory results (particularly on the Ventana Benchmark platform), and mAb clone TF3H8-1 (RTU 760-2507, Ventana) performed poorly. Optimal results could only be obtained with use of HIER in an alkaline buffer. The tissue material was more challenging than in previous runs, but the causes of sufficient and insufficient results are basically the same.

CK20: 284 participants, 92% sufficient, 62% optimal. Efficient HIER is recommended, proteolytic pretreatment generally gives a lower pass rate.

CK-PAN: 275 participants, 72% achieved a sufficient mark, 48% optimal. For Ab cocktails containing AE1/AE3 HIER is mandatory. mAb MNF116 requires proteolytic pretreatment but the clone performs less well than AE1/AE3.

CyD1: 257 participants, 94% sufficient, 54% optimal. Both rmAb clones EP12 and SP4 can be recommended. However, SP4 performed less optimal on the Bond platform.

SOX11: 79 participants, 66% sufficient, 27% optimal. mAb clones MRQ-58 and SOX11-C1 can both be recommended. However, clone MRQ-58 in RTU format from Ventana (760-4888) works better with OptiView than UltraView (that is receommended by the producer).

Run 48, B22, H10 opens for protocol submission on 12th August.

July 11th 2016
Mogens Vyberg
Scheme director