Alternative content

April 2008, VOL 11 Connection IHC STANDARDIZATION AROUND THE WORLD In this issue QA with Pathologists on I mmunohisto chemistr y Standardization Profiles, inter views, opinions and ar ticles from leading researchers April 2008, VOL 11 Connection IHC STANDARDIZATION AROUND THE WORLD In this issue QA with Pathologists on I mmunohisto chemistr y Standardization Profiles, inter views, opinions and ar ticles from leading researchers Contents APRIL 2008, VOL 11 2 Editorial 3 Featured Laboratory Osamura Laboratory The George L. Kumar, PhD 4 QA-Ask the Experts IHC Standardization Around the World Anthony S-Y Leong, MB, BS, MD, FRCPath, FCAP, FASCP 4 Fernando Soares, MD, PhD 9 ryan R. Hewlett, ART,MLT B 12 Prod Editorial Immunohistochemistry (IHC) Standardization Around the World George L. Kumar, PhD Managing Editor Welcome to another informative issue of Connection, our special issue devoted to ?IHC Standardization Around the World? This issue . includes opinions, articles and questions and answ Featured Laboratory The Osamura Laboratory Laboratory headed by Prof. Robert Yoshiyuki Osamura, MD, PhD Department of Pathology/Diagnostic Molecular Pathology Laboratory, Tokai University School of Medicine. Isehara Campus, Kanagawa prefecture, Japan. Back row: Hitoshi Itoh, CMIAC, Nobuaki Kato, C QA Ask the Experts: On Immunohistochemistry (IHC) Standardization Professor Leong is Medical Director of Hunter Area Pathology Service and Chair of Anatomical Pathology, University of Newcastle in Australia. He is also Adjunct Professor at Peking University and has held visiting professorships at u time interval between removal of tissue and immersion in fixative, the temperature of tissue storage and the site of tissue sampling, the latter including the variability of antigen preservation between necrotic and nonnecrotic tissues. Pre-analytical variables lie in the domain of the surgeon and c Connection: How would you like to standardize IHC in Australia? As discussed above, standardization will not be possible, if we cannot control or influence the pre-analytical factors. We would be better off developing a standard control that can be inserted into the test slide. You say, ?standard Connection: Antigen retrieval is essential in immunohistochemistry (IHC) in order to restore epitopes. There are a large number of devices which can be used to achieve this aim (microwaves, rice steamers, pressure cookers, autoclaves and commercial de-cloaking devices). However, the variability of r Connection: Is a universal image analysis system feasible? No, not until we standardize the all important pre-analytical variables discussed above. Connection: In your opinion, what is the biggest hurdle for standardization in image analysis? I do not believe it will solve the major problems. My rea QA Fernando Soares, MD, PhD University of São Paulo, São Paulo, Brazil Dr. Fernando Soares is Full Professor at the University of São Paulo, São Paulo, Brazil, and Head of the Department of Anatomic Pathology of the Research and Medical Hospital A.C. Camargo, São Paulo, Brazil. He also serves Connection: In your opinion, what is the biggest hurdle for standardization? Probably education in laboratory practices. Most of the pathologists are very well trained in diagnosis, but lack bench training and training in laboratory practices. Also, costs associated with training is an important iss There are no standards in Latin America. We always follow the manufacturer?s protocol during our first test. We have tested different protocols of antigen retrieval in our breast cancer panel, and we always test different protocols for each antibody in our laboratory. Therefore, an antibody to be QA Bryan R. Hewlett, ART, MLT Consultant Technologist to the Anatomic Pathology EQA program of Ontario Quality Management ProgramLaboratory Services (QMP-LS) Canada Bryan R. Hewlett, ART, MLT. Bryan received his initial and specialist training as a Medical Laboratory Technologist in Nottingham, U.K. There is no standardization of AR steps ? circumstances, it would be impossible to and, under a the present fixation provide single standard ... ? Connection: How well is the ?pre-fixation? process standardized? Is it at all possible to standardize this process? It is not standardized. If Connection: How would you rate European (UK NEQAS, NordiQC) and US IHC standards to Canadian IHC standards? There are no standards. IHC is a highly complex procedure no part of that complexity can be ignored. It is possible to produce equivalent results using a variety of methods. This is what needs QA Prof. Chen Jie Prof. Cui Quancai Peking Union Mediacl College Hospital, Beijing, China Prof. Chen Jie Professor of Pathology and Vice President of the Peking Union Medical College (PUMC) Hospital, Professor Chen Jie is also the President of the Chinese Society of Pathology and Deputy Presiden Connection: According to Goldstein et al. Appl Immunohistochem Mol Morphol 2007;15:124?133 ?Immunohistochemistry (IHC) continues to suffer from variable consistency, poor reproducibility, quality assurance disparities, and the lack of standardization resulting in poor concordance, validation, an Connection: Is it true that a particular histology feature may be better demonstrated by other fixatives, such as Glyo-Fixx for nuclear features and lymphocyte appearance and Omnifix for cytoplasmic detail (Arch Pathol Lab Med 2005;129:502-506)? Would you recommend these fixatives in the future? Are Connection: What constitutes standardization of image analysis as applied to immunohistochemistry (IHC)? Can we have a standardized procedure to interpret IHC assays around the world? Highquality IHC slides and a well-trained technician on a perfect image analysis device. Yes. We recommend using a s QA Dr. Tanuja Manjanath Shet Dr. Vani Parmar Tata Memorial Hospital, Mumbai, India Tanuja Manjanath Shet, MD, DPB, DTM, DNB Dr. Vani Parmar MBBS, MS, DNB Dr. Shet is a Professor and Pathologist at the Tata Memorial Hospital, Mumbai, a large cancer center in India. She is a Surgical Pathologis ? ... the biggest hurdle in India is suboptimal fixation and processing of tissues. ? Though I agree that IHC has the limitations mentioned in the above article, I do believe these can be surpassed. Just as it is important to do IHC it is equally important to validate IHC. Every signing patholog Connection: Is it true that a particular histology feature may be better demonstrated by other fixatives such as Glyo-Fixx for nuclear features and lymphocyte appearance and Omnifix for cytoplasmic detail (Arch Pathol Lab Med 2005;129:502-506)? Would you recommend these fixatives in the future? Are Connection: Can you comment on the internal and external quality control (EQC) procedures followed in India? Specifically, which agency in India is involved in EQC? Cf. 4 and 5. Connection: What are the major benefits of EQC? All labs work toward their best, but they really don?t know where they s QA Prof. Robert Yoshiyuki Osamura Department of Pathology, Tokai University School of Medicine Japan. Isehara Campus, Japan ? ... IHC Standardization in pathology itself has not been progressed sufficiently and the situation is recognized as a big problem ... Degree: MD, PhD Current p Connection: In your opinion, what is the biggest hurdle for standardization? The biggest hurdle for standardization is considered to lie in the development of a nationwide system to manage the process. There is also a limited budget for employing people for quality control and managing the process o the standardization of image ? ... appropriate for pre-screening analysis is of the staining quality. ? ? Allred DC, Harvey JM, Berardo M, Clark GM: Prognostic and predictive factors in breast cancer by immunohistochemical analysis. Mod Pathol 11:155-168, 1998. Connection: Antigen retr Connection: Why is standardization of image analysis in diagnostic pathology important? Because the results are directly related to the most appropriate ?diagnosis and treatment? of a patient. Connection: What are your recommendations for standardizing image analysis in an anatomic pathology lab QA James F. Happel, DLM (ASCP), HTL Technical Director of Surgical Pathology, Research and Development at Massachusetts General Hospital. Boston, Massachusetts Mr. Happel is certified by the American Society for Clinical Pathology as a Histotechnologist and as a Diplomate in Laboratory Manage Connection: United Kingdom National External Quality Assessment Service (UK NEQAS) helps to ensure that clinical laboratory test results are accurate, reliable and comparable wherever they are produced. Similarly, Nordic Immunohistochemical Quality Control (NordiQC) an independent, scientific, non-p Connection: The American Society of Clinical Oncology (ASCO) and the College of American Pathologists (CAP) convened an expert panel to develop a guideline to improve the accuracy of HER2 testing in invasive breast cancer. The ASCO/CAP Guideline is also followed by Canadian pathologists with some mo would recommend that standardization ? I begin with identifying a reliable and trustworthy source ... ? Connection: What are the major benefits of EQC? Some of the major benefits of EQC are verification of the overall process and accuracy of your IHC stains. Applying the use of the double-b Interview Immunohistochemistry for Oestrogen and Progesterone Receptors Dr. Andrew Lee Consultant Histopathologist, Nottingham City Hospital, UK Connection: Can oestrogen receptor status of invasive carcinoma of the breast be reliably assessed on core needle biopsy (CNB)? In a recent au Connection: What is the difference between the H score and the Allred score? Which is better? What do you prefer? Why are you not using the Allred scoring system? Both methods combine assessment of percentage of tumour cell nuclei staining and the intensity of staining. The H score is the sum of the Connection: Can you comment on the burden in the laboratory, if one changes from a current ER/PR assay to a standardised ER/PR assay? Important decisions in standardising these assays are the choice of fixation, processing, pre-treatment, primary antibody and visualisation method. These choices can Opinion Interview IHC Standardization: A Dako Perspective Dr. Ole Rasmussen RD Director, Systems Development Dako Denmark A/S Produktionsvej 42 DK-2600 Glostrup Denmark Connection: Can you mention a few key factors for promoting standardization of IHC in Europe? I think that quality assessment or Connection: Dako has developed Readyto-Use Antibodies for in vitro diagnostic applications. How is this going to facilitate standardization? We have great expectations as far as regards our new Ready-to-Use (RTU) Series of Antibodies. This series includes 65 new antibodies ? each of which are pre- Articles UK NEQAS-ICC ISH: Historical perspective, current role, future directions Andy Dodson, M. Phil. CSci, FIMBS Started his career in medical science at St. George?s Hospital London in 1981. After qualifying, he went on to work in research for the British Heart Foundation for four years. UK NEQAS-ICC in the 1990s In his first year as Scheme Organiser, Keith oversaw the transition to subscription-based financing (previously the Scheme had been free to use as it was funded by top-slicing). He also saw the membership increase to more than 200. It quickly became obvious that a full-time The application could be argued to represent a field change in terms of the rigour with which the analysis is required to be done. It certainly offered a new challenge to Keith Miller and Tony Rhodes as their solution was to pioneer the use of cell-line material for the external quality assessment o Assessment teams consist of four assessors, who view slides around a multi-headed microscope and score independently. While the method has remained pretty much unchanged from the very first assessment, the mechanism by which it is achieved has been improved significantly with the introduction under The archive which UK NEQAS holds, both in terms of stained slides and methodological data, must surely be unrivalled and represents an invaluable research resource. While it would be impossible for UK NEQAS to exploit this stream of data to its full potential as it must devote the majority of its re For Immunocytochemistry and FISH RESULT: RUN 80L SLIDE: NEQAS Laboratory No: XXX Mr. A. Scientist Department of Histopathology Anytown Hospital Anytown SCHEME: Lymphoma August 2007 Scheme Assessor Assessor1 Assessor2 Assessor3 Assessor4 Lymphoma Mark 3 3 2 3 Assessment Code 80L Comment 2 Antibo Figure 6. Feedback on results has always been given high priority, and for many years this has been facilitated by the publication of a Scheme journal which is freely available to participants. In its original incarnation in 1992, the Journal (left) ran to eight pages and contained three photomicrog a b c d Figure 7. The antigen chosen by Gerry Reynolds for his very first assessment run was kappa light chain. The choice was either inspired or very fortuitous (knowing Gerry, I think the former is more likely). Successful demonstration of this antigen requires careful attention to detail at e Bibliography Selected UK NEQAS-ICC ISH papers. Ibrahim M, Dodson AR, Barnett S, Fish D, Jasani B, Miller K. Potential for false positive staining with a rabbit monoclonal antibody to progesterone receptor (SP2): findings of the UK National External Quality Assessment Scheme for Immunocytochemistry Articles Nordic Immunohistochemical Quality Control (NordiQC) An Organization for External Quality Assurance Mogens Vyberg, MD Institute of Pathology. Aalborg Hospital Aarhus University Hospital Denmark Søren Nielsen, HT, CT Institute of Pathology Aalborg Hospital Aalborg University Hospital Denm parameters (i.e. results interpretation and reporting) (4, 5). In an EQA setting, by circulating serial sections to a number of laboratories to be stained for specific antigenic markers and assessing them in a standardized and objective way, the preand post-analytical parameters influencing the resu CD79a (Fig. 2) Among 112 laboratories submitting stains in the latest run, most used mAb clone JCB117 and obtained sufficient results in 85%. In contrast, all of the six laboratories using mAb clone HM57 obtained poor marks, due to insufficient staining of neoplasms (Fig. 2) Even the Dako Web site s References 1. Rhodes A, Jasani B, Barnes DM, Bobrow LG, Miller KD. Reliability of immuno-histochemical demonstration of oestrogen receptors in routine practice: interlaboratory variance in the sensitivity of detection and evaluation of scoring systems. Clin Pathol 2000;53(2):125-30 2.Varma M, Ber Fig. 2. CD79a A. Optimal CD79a staining of the tonsil using the monoclonal antibody (mAb) clone JCB117. The mantle zone B-cells show an intense staining, while the germinal centre B-cells show a moderate staining. B. CD79a staining of the tonsil ? same field as in (A) using an insufficient proto Standardization of Ki-67 Immunohistochemical Staining for Diagnosing Grade of Gastrointestinal Stromal Tumor (GIST) A report of the research regarding the establishment of pathological diagnostic criteria in order to standardize cancer diagnosis and treatment. Supported by a grant-in-aid for cancer was conducted in 49 GIST cases. The concordance rate for the evaluation results at three laboratories was 90% or higher and the k value was 0.80 or more among all laboratories.Thus, essentially equal staining results were obtained in these three laboratories. It was thus confirmed that the STD metho CB pH6 a b c d TE pH9 e f g h Autoclave 121° C/10 min Water bath 95° C/40 min Microwave 5 min x 3 cycle Microwave 10 min x 3 cycle Figure 1. Comparison of staining intensity among eight different heat-induced epitope retrieval conditions. Anti-Ki-67 (Clone MIB-1) antibody was used 50 Table 1. Correlation between NCC and STD methods R 2 =0.9483 Categories of proportion NCC Method STD Method 40 Opinions Importance of Standardization for Predictive Prognosis David J. Dabbs, MD Chief of Pathology, Magee-Women?s Hospital University of Pittsburgh School of Medicine Estrogen and progesterone receptors have both prognostic and predictive value, but the predictive value significantly outwe is documented and serves as a surrogate marker for the initial exposure to formalin. Since the first two stations in our tissue processors are formalin, comprising two hours, six additional hours of formalin exposure are required before going onto our tissue processors. The surrogate endpoint out of Opinions The New Era for ER and PRIt?s time to Standardize! Dr. Ian Ellis, B.Med.Sci. BM, MS, FRCpath Molecular Medical Sciences, University of Nottingham Departments of Histopathology, Nottingham City Hospital NHS Trust Biodata Professor Ian O. Ellis has been involved in the practice of path et al 2001). The main reason for false-negative results is due to inefficient heat-induced epitope (antigen) retrieval (Barnett S 2007). Additional reasons for differences between laboratories are variations in: ? primary antibodies ? dilutions of primary antibodies ? incubation temperature Standardization of HER2 TestingInconsistency Raises Questions Opinions Sunil S. Badve, MD, FRCPath Clarian Pathology Laboratory, Indiana University School of Medicine An expert in the field of breast pathology, Dr. Badve is Associate Professor in the Department of Pathology and Laboratory Medic rence seen in these trials is in the order of 50%. This is the major reason for all the excitement associated with this drug. It is thus important for us pathologists to get the testing right. The principal testing methods used for determination of HER2 status are immunohistochemistry for protein ov which now requires expression of HER2 by at least 30% of tumor cells (instead of 10%). It has also recognized that FISH and IHC are equivalent in their efficacy in identifying patients who are likely to be responders to trastuzumab. They have also generated an equivocal category of FISH interpretati IHC CONSENSUS MEETING, JANUARY 27 2008, SANTA BARBARA, CA, USA , IHC CONSENSUS MEETING, JANUARY 27, 2008, SANTA BARBARA, CA, USA SPONSORED BY DAKO NORTH AMERICA, INC. To further the development of quality assurance and standardization procedures in immunohistochemistry, several leaders in the fiel Richard Cartun, PhD, Sunil Badve, MD Jon Askaa, PhD Søren Nielsen, HT, CT, Mogens Vyberg, MD Elizabeth Hammond, MD David Dabbs, MD, Roscoe Atkins, MD Neal Goldstein, MD, David Hicks, MD Back rowfrom left to right: Bryan Hewlett ART, MLT, Alton Floyd PhD: Alvin Martin MD, Todd Barry MD, PhD. Fro Dako Abstracts Abstracts presented at the 30th Annual San Antonio Breast Cancer Symposium December 13-16, 2007 San Antonio, Texas, USA Relationship between ESR1 copy number and ER expression in the DBCG 89D trial Amplification of ESR1 may predict resistance to adjuvant tamoxifen in postmenopausal p Dako Abstracts Amplification of ESR1 may predict resistance to adjuvant tamoxifen in postmenopausal patients with hormone receptor positive breast cancer Ejlertsen B, Nielsen KV, Rasmussen BB, Balslev E, Müller S, Møller S, Mouridsen HT. Rigshospitalet, Copenhagen, Denmark; Dako A/S, Glostrup, De Dako Abstracts Abstracts presented at the ?United States and Canadian Academy of Pathology (USCAP) Annual Meeting. March 1-7, 2008 Denver, Colorado, USA IMP3 Is an Extremely Poor Prognostic Marker for Ovarian Clear-Cell Carcinoma Köbel M, Xu H, Bourne PA, Spaulding BO, Huntsman D, Gilks CB. Vanc Dako Abstracts Metastatic Pancreatic Endocrine Tumors in the Liver Express KOC Briones AJ, Bourne PA, Spaulding BO, Lu D, Qu Z, Fischer-Colbrie R, Wang HL, Xu H. University of Rochester Medical Center, Rochester; Dako North America, Carpinteria; Washington University School of Medicine, St. Louis; Dako Abstracts Merkel Cell Carcinomas Express K Homology Domain Containing Protein Overexpressed in Cancer like Small Cell Carcinomas Pryor JG, Simon RA, Bourne PA, Spaulding BO, Scott GA, Xu H. University of Rochester Medical Center, Rochester, NY; Dako North America, Carpinteria, CA Background K Dako Abstracts Immunohistochemical Analysis of KOC/IMP3 in Malignant Pleural Mesothelioma Xu H, Simon R, Bourne PA, Spaulding BO, Wang HL. University of Rochester Medical Center, Rochester, NY; Dako North America, Carpinteria, CA; Cedars-Sinai Medical Center, Los Angeles, CA Background K homology Dako Abstracts KOC, TTF-1 and CDX2 Discriminate Small-Cell Carcinoma from Carcinoid and Pancreatic Endocrine Tumor Metastasized to the Liver Xu H, Briones AJ, Bourne PA, Spaulding BO, Lu D, Fischer-Colbrie R, Qu Z, Wang HL. University of Rochester Medical Center, Rochester; Dako North America, Carp Dako Publications Publications Co-authored by Dako: In Press Li L, Xu H, Spaulding B, Cheng L, Simon R, Jorge L. Yao, P. Anthony di Sant? gnese A, Bourne A PA and Huang J. Expression of KOC (K-domain protein overexpressed in human cancer) in benign urothelium and urothelial tumors. Human Pathol Dako Meetings 2007 National Society for Histotechnology Meeting. Denver, CO NSH workshop attendees listening to David E Muirhead (picture on the left), Assistant Professor of Pathology and Microbiology The Nebraska Medical Center and Paula B Williams (picture on the right), Lead Histotechnologist New Premium Quality Concentrated Antibodies New Products Your current and future needs drive the continuing expansion and improvement of Dako?s antibody selection. With the current addition of five new premium quality concentrated antibodies and a recent launch of seven others, Dako?s portfoli Targeting Colorectal Adenocarcinomas, Anti-TIMP-1 and Anti-Villin Monoclonal Mouse Anti-Human Tissue Inhibitor of Metalloproteinases 1 Clone: VT7 Code: M7293 (0.2 mL) ? In Vitro Diagnostics ? Frozen ? Formalin ? HIER Tissue Inhibitor of Metalloproteinases 1 (TIMP-1) is capable of inhibiting Dako?s FLEX Ready-to-Use Concept Enhance performance of your laboratory by running FLEX Ready-to-Use antibodies on your Dako instruments. New Products EnVisionTM FLEX+ increases signal of Anti-TTF-1, clone 8G7G3/1 labelling in lung adenocarcinoma EnVisionTM FLEX+, increases signal amplification 2008 Product Catalog Available Now! The catalog features more than 170 new products, including the Autostainer Plus Link and the True Positive ID which connect the pathology laboratory using the innovative Dako Link software. Together with the Pre-Treatment (PT) Link Module and Laboratory Informatio