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Belgian NordiQC Workshop 13-15 June 2018

The 1st NordiQC Workshop in Brügge, 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: 60.

Programme and registration available in September '17.

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

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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