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1 Zyto_Facts News for pathology and immunohistochemistry +++Newsflash +++ Newsflash++ Speaking to you IHC algorithm poster now available in English. You can download the poster directly from our homepage or ask your local supplier for the printed version. CEO Dr. K. Debel CEO T. Dittmer CEO K. Weyrauch ALK/p80 for detecting non-small cell lung cancer with ALK rearrangement now available at Zytomed Systems 25th European Congress of Pathology of the ESP, 31 Aug 04 Sept 2013, Centro de Congressos, Lisbon, Portugal The 8th Asia Pacific IAP Congress, 5 8 Sept 2013, Busan Exhibition & Convention Centre, Busan, Korea 63rd National Congress of the Italian Association of Clinical Pathology and Molecular Medicine, Sept 2013, Perugia, Italy XXIX Congress of the Latin American Society of Pathology, Oct 2013, TBA, Oaxaca, Mexico Joint Annual Meeting of the Swiss and Austrian Societies of Pathology (SG- Path/SSPath and ÖGP), 7 9 Nov 2013, TRAFO Baden, Baden, Switzerland Carrefour Pathologie, Nov 2013, CNIT, Paris La Défense, France USCAP 2014 Annual Meeting, 1 7 March 2014, San Diego CA, USA 3rd Pannonia Congress of Pathology, May 2014, Bled, Slovenia PathSoc Joint Meeting with the European Society of Pathology, 31 Aug 4 Sept 2014, ExCeL, London, United Kingdom Welcome to our 2013 issue of Zyto_Facts featuring information about new antibodies and technical issues! We would like to draw your attention to our new antibody against the SOX-11 protein and also to our excellent PAX-8 antibody this marker is becoming more and more important for ovarian and kidney cell carcinomas. Quality control in immunohistochemistry is a constant challenge for all IHC labs. The section Focus: lab work targets that challenge, focussing especially on pre-analytics which receive little attention in most pathology labs. PAX-8 A specific and sensitive marker for ovarian and kidney cell carcinomas Pax-8 is relatively new on the scene, but I have found it to be one of the most useful and highly valued markers for addressing the problem of metastatic carcinoma of unknown primary, where it can be used as a marker of thyroid carcinomas, female genital tract carcinomas, and renal cell carcinomas. This statement by Dr. Rodney T. Miller, Director of PAX-8 as a marker for ovarian carcinomas and their metastases Recent publications describe PAX-8 as a useful marker for ovarian carcinomas [2,3]. According to Tacha et al. [4] PAX-8 can be detected via immunostain in 79 % of all ovarian carcinomas. Carcinomas of the breast and lung do not express PAX-8 [2]. Therefore positive PAX-8 staining can be very helpful to confirm the ovary or fallopian tube as the primary tumour site. This is especially helpful as both ovarian and breast carcinomas share similar morphologic features. On the other hand, an invasive micropapillary carcinoma found in the ovary and tested negatively for PAX-8 most probably has its origin in another organ. With compliments, your Zytomed Systems Management Team Immunohistochemistry at the ProPath Laboratory in Dallas, Texas [1] highlights the extremely useful features of PAX-8. Zytomed Systems offers an excellent antibody suitable for FFPE tissue sections to detect this protein. The transcription factor PAX-8 is a nuclear protein involved in the development of the thyroid gland, kidney and Müllerian system. PAX-8 staining on ovarian carcinoma (RBK047-05)
2 Expression of PAX-8 in ovarian and breast carcinomas (according to Nonaka et al. [3]) Tumour PAX-8 Positivity Serous papillary ovarian carcinoma (n = 84) 96 % Endometrial ovarian carcinoma (n = 18) 89 % Mucinous ovarian carcinoma (n = 12) 8 % Clear cell ovarian carcinoma (n = 10) 100 % Breast carcinoma, ductal (n = 178) 0 % Breast carcinoma, lobular (n = 65) 0 % PAX-8 as a marker for kidney cell carcinomas and their metastases Besides its value as an ovarian carcinoma marker, PAX-8 is also a helpful marker for kidney cell carcinomas and their metastases. Ozcan et al. describe PAX-8 as a diagnostically useful marker for both primary and metastatic renal neoplasms of a large variety of histologic types [5]. In this study PAX-8 shows a higher positivity than PAX-2 for renal cell carcinomas (RCC). Rodney T. Miller also describes PAX-8 to be the superior marker not only to PAX-2 but also to the long-established RCC: To be honest, I think both RCC and PAX-2 are overrated as markers of kidney tumors, particularly since PAX-8 became available as a kidney marker [1]. The high sensitivity for PAX-8 on RCC was also confirmed by Tong et al. and by Knoepp et al. on cytological specimens [6,7]. Roughly 85 % of all kidney tumours are renal cell carcinomas. The most common subtype of RCC is clear cell RCC which accounts for about 83 % of all RCC [8]. Very rarely, PAX-8 negative RCC metastases can be positive for PAX-2. Therefore Ozcan et al. suggest retaining PAX-2 in the diagnostic panel for kidney tumours [5]. PAX-8 staining on clear cell RCC (RBK047-05) Expression of PAX-8 and PAX-2 in kidney cell carcinomas Tumour PAX-8 Positivity PAX-2 Positivity Clear cell RCC (n = 95) 97 % 95 % Papillary RCC (n = 38) 100 % 76 % Chromophobe RCC (n = 25) 88 % 56 % Oncocytoma (n = 13) 85 % 54 % PAX-2 staining on clear-cell RCC (RBK013-05) Collecting renal tube RCC (n = 7) 71 % 43 % RCC metastases (n = 99) 89 % 76 % PAX-2 staining on clear-cell RCC (RBK013-05) Product information PAX-8 Clone: polyclonal Ready-to-use 6 ml RBG047 1:25 1: ml RBK PAX-2 Clone: polyclonal 1:20 1: ml RBK HIER: Heat Induced Epitope Retrieval Method: P = Immunohistochemistry on FFPE tissue sections F = Immunohistochemistry on frozen sections
3 Bibliography [1] Miller RT. Immunohistochemistry in the Diagnosis of Metastatic Carcinoma of Unknown Primary Origin. American Academy of Oral and Maxillofacial Pathology Annual Meeting. San Juan, Puerto Rico, April 30, 2011 [2] Laury AR et al. A comprehensive analysis of PAX8 expression in human epithelial tumors. Am J Surg Pathol 35: , 2011 [3] Nonaka D et al. Expression of PAX8 as a useful marker in distinguishing ovarian carcinomas from mammary carcinomas. Am J Surg Pathol 32: , 2008 [4] Tacha D et al. Expression of PAX8 in normal and neoplastic tissues: a comprehensive immunohistochemical study. Appl Immunohistochem Mol Morphol 19: , 2011 [5] Ozcan A et al. PAX2 and PAX8 expression in primary and metastatic renal tumors: a comprehensive comparison. Arch Pathol Lab Med 136: , 2012 [6] Tong G-X et al. Expression of PAX8 in normal and neoplastic renal tissues: an immunohistochemical study. Mod Pathol 22: , 2009 [7] Knoepp SM et al. Utility of PAX8 and PAX2 immunohistochemistry in the identification of renal cell carcinoma in diagnostic cytology. Diagn Cytopathol 40: , 2012 [8] Motzer RJ et al. Renal-Cell Carcinoma. N Engl J Med 335: , 1996 SOX-11 A new and sensitive marker for Cyclin D1 negative mantle cell lymphomas SOX-11 staining on mantle cell lymphoma (MSK095-05) In addition to an excellent, globally established antibody against Cyclin D1 (clone SP4), Zytomed Systems offers an antibody against SOX-11 a marker especially helpful for detecting Cyclin D1-negative mantle cell lymphomas. That is a difficult task because they may resemble other small-cell B-cell lymphomas morphologically and phenotypically. Since patients with small-cell B-cell lymphoma mimicking mantle cell lymphomas have a significantly better outcome than those with true mantle cell lymphoma, it is important to use reliable biomarkers that identify true mantle cell lymphomas. SOX-11 staining can be very useful for differentiating between these two types of lymphoma % of all mature B-cell neoplasias are mantle cell lymphomas characterised by overexpression of Cyclin D1. Roughly 5 10 % of these mantle cell lymphomas are negative for Cyclin D1. According to Mozos et al. up to 93 % of all mantle cell lymphomas, including those which are negative for Cyclin D1, express SOX-11 [1]. Other publications also describe the high sensitivity of SOX-11 for mantle cell lymphomas [2,3]. Thus, SOX-11 can be very helpful in haematopathological differential diagnosis. As SOX-11 is a transcription factor, its expression is nuclear. In addition to mantle cell lymphoma, lymphoblastic lymphomas (80 %) and Burkitt lymphomas (20 33 %) are positive for SOX-11 [4,5]. SOX-11 is not detectable in CD5-positive diffuse large-cell B-cell lymphomas [5]. Bibliography [1] Mozos A et al. SOX11 expression is highly specific for mantle cell lymphoma and identifies the cyclin D1-negative subtype. Haematologica 94: , 2009 [2] Chen YH et al. Nuclear expression of SOX11 is highly associated with mantle cell lymphoma but is independent of t(11;14)(q13;q32) in non-mantle cell B- cell neoplasms. Mod Pathol 23: , 2010 Product information SOX-11 Clone: ZSX11 Cyclin D1 Clone: SP4, MS, RT or EDTA ph 9.0 Ready-to-use 6 ml MSG095 1:25 1: ml MSK Ready-to-use 6 ml RBG025 1:25 1: ml RBK HIER: Heat Induced Epitope Retrieval Method: P = Immunohistochemistry on FFPE tissue sections F = Immunohistochemistry on frozen sections [3] Lu J and Chang KL. Practical Immunohistochemistry in Hematopathology: A Review of Useful Antibodies for Diagnosis. Adv Anat Pathol 18: , 2011 [4] Dictor M et al. Strong lymphoid nuclear expression of SOX11 transcription factor defines lymphoblastic neoplasms, mantle cell lymphoma and Burkitt s lymphoma. Haematologica 94: , 2009 [5] Zeng W et al. Cyclin D1-negative blastoid mantle cell lymphoma identified by SOX11 expression. Am J Surg Pathol 36: , 2012
4 Zyto_Facts News for pathology and immunohistochemistry Focus: lab work Quality control in immunohistochemistry a constant challenge for every lab Pathology laboratories produce many reports each day reports which the lives of patients depend on. Therefore each laboratory needs to establish its own quality control procedures to ensure reliable and constant results in its dayto-day work. Simply doing positive and negative controls - is this sufficient quality control in immunohistochemistry? Of course these controls are necessary, but quality means more. The first thing that surprises everybody when starting certification or accreditation procedures is the importance of documentation. Documentation doesn t mean simply writing down which antibody is used on which tissue and documenting the patient data. Documentation also means recording the IHC protocol employed, writing down which lot numbers were used for every reagent, and also documenting the entire process of pre-analytics. Thinking about pre-analytics is often overlooked. For a successful immunohistochemical stain, it is important to know about pre-analytic procedures because they influence the results massively. What happened in the operating theatre? is an important question for the pathologist. It certainly does matter whether the patient material was put into formalin immediately or kept at room temperature for 5 hours first. In our Zyto_Facts 1_2012 we described some critical points during fixation. and chromogenic substrate do not produce falsepositive stains. Positive control materials can be tissues from the lab itself which tested positive for the antigen in question. Zytomed Systems offers a Receptor Control Block containing cell lines with different expression levels of ER, PR and HER2 for quality control purposes. Some countries suggest using on-slide positive controls for certain markers. In Germany for example, accredited laboratories must perform on-slide controls for predictive or prognostic markers. Participating in national and international quality control schemes is an excellent way to demonstrate the quality of immunohistochemistry of a laboratory. Find more about this topic on the last page of this newsletter. All in all, quality control in immunohistochemistry (and not only there) is a never-ending process which needs constant attention. Although it takes time and effort, quality control is necessary to ensure the best possible pathological report for the benefit of the patient. This cartoon is kindly provided by Ursula Hofmann. She lives and works in Berlin and would be pleased to bring your ideas into coloured life. If you would like to contact her please use here web page or ask Zytomed Systems. We will be glad to assist you. Also, the entire procedure of dehydrating the tissue until it becomes a paraffin block needs to be documented. An IHC laboratory working according to GLP will also standardise the thickness of the cut sections and the pre-treatment procedure. Standardising the entire pre-analytic procedure is always worthwhile. It is important in order to obtain reliable results in immunohistochemistry. Let s come back to positive and negative controls. There should be one positive control for each of the antibodies during every staining run, no matter if it is done manually or on an immunostainer. A positive stain on this control tissue will confirm the stain on the patient tissue. Negative controls will make sure your IHC detection system
5 Zytomed Systems laboratory and reagents Constantly excellent results in NordiQC scheme For many years Zytomed Systems has been participating in national and international quality control schemes for immunohistochemistry. These quality control schemes are an excellent opportunity for us to test the performance of our laboratory and reagents. We participate in the German Quality Initiative Pathology (QuIP) as well as in the Nordic immunohistochemical Quality Control (NordiQC), which is operated by the Institute of Pathology, Aalborg Hospital, Denmark. In recent years, our laboratory participated in the NordiQC runs with roughly 30 different antibodies and many detection reagents. Our immunohistochemical stains were rated optimal 13 times and good 14 times. The following tables list a few of the antibodies and reagents which were awarded the result optimal. CDX2 (RBK019) on colon carcinoma biopsy Primary antibodies CD20 Clone: L26 CD30 (Ki-1) Clone: Ber-H2 CD45 Clone: 2B11&PD7/26 CDX2 Clone: EPR2764Y Cyclin D1 Clone: SP4 Cytokeratin Pan Clone: AE1&AE3 Desmin Clone: D33 Estrogen Receptor Clone: 1D5 P None, MS, RT, MS, RT, DG, MS, RT, MS, RT, DG, PG or EDTA ph 9.0 or Pepsin Ready-to-use 6 ml MSG ml MSK ml MSK008 Ready-to-use 6 ml MSG061 1:40 1: ml MSK ml MSK061 Ready-to-use 6 ml MSG ml MSK ml MSK054 Ready-to-use 6 ml RBG ml RBK ml RBK019 6 ml RBG ml RBK Ready-to-use 6 ml MSG ml MSK ml MSK019 Ready-to-use 6 ml MSG053 1:25 1: ml MSK ml MSK053 Ready-to-use 6 ml MSG ml MSK ml MSK001 Podoplanin (MSK057) on mesothelioma
6 Zyto_Facts News for pathology and immunohistochemistry HER2 Clone: SP3 Ready-to-use 6 ml RBG ml RBK ml RBK026 Podoplanin Clone: D2-40 Ready-to-use 6 ml MSG ml MSK ml MSK057 Vimentin Clone: V9, MS, RT, DG, PG (optional) Ready-to-use 6 ml MSG ml MSK ml MSK023 Detection Reagents, chromogenic substrates and accessories Description Volume Cat. No. ZytoChem Plus HRP Polymer Kit anti-mouse/rabbit 100 ml POLHRP-100 Antibody Diluent 500 ml ZUC DAB Substrate Kit 500 ml DAB530 HIER Citrate Buffer ph 6.0 (10X) 500 ml ZUC Tris Wash Buffer (20 X), TBS 500 ml ZUC Peroxide Block 500 ml ZUC Your local contact: contact ZYTOMED SYSTEMS GmbH Anhaltinerstraße Berlin Germany Fon Fax info@zytomed-systems.de Juni-2013 NL_E_1_2013
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