Immunohistochemistry with the anti-BRAF V600E (VE1) antibody: impact of pre-analytical conditions and concordance with DNA sequencing in colorectal and papillary thyroid carcinoma

Summary The most common of all activating BRAF mutations (T1799A) leads to a substitution of valine (V) to glutamic acid (E) at the position 600 of the amino acid sequence. The major goal of this study was to compare detection of the BRAF V600E mutation by DNA sequencing with immunohistochemistry (IHC) using the anti-BRAF V600E (VE1) antibody. Archival formalin fixed, paraffin embedded tissues from 352 patients with colon adenocarcinoma (n = 279) and papillary thyroid carcinoma (n = 73) were evaluated for the BRAF V600E mutation by sequencing and IHC. The discordant cases were re-evaluated by repeat IHC, SNaPshot and next-generation sequencing (NGS). Furthermore, the effect of pre-analytical variables on the utility of this antibody was evaluated in two xenograft mouse models. After resolving 15 initially discordant cases, 212 cases were negative for the BRAF V600E mutation by IHC. Of these, 210 cases (99.1%) were also negative by sequencing and two cases (0.9%) remained discordant. Of the 140 cases that were IHC positive for BRAF V600E, 138 cases were confirmed by sequencing (98.6%) and two cases remained discordant (1.4%). Overall, the negative predictive value was 99.1%, positive predictive value 98.6%, sensitivity 98.6%, specificity 99.1% and overall percentage agreement 98.9% (348/352 cases). Tissue fixation studies indicated that tissues should be fixed for 12–24 h within 2 h of tissue collection with 10% neutral buffered formalin.

[1]  P. Gibbs,et al.  A mutant BRAF V600E-specific immunohistochemical assay: correlation with molecular mutation status and clinical outcome in colorectal cancer , 2014, Targeted Oncology.

[2]  P. Waring,et al.  The prognostic significance of the BRAFV600E mutation in papillary thyroid carcinoma detected by mutation-specific immunohistochemistry , 2013, Pathology.

[3]  F. Deng,et al.  Clinical utility of immunohistochemistry for the detection of the BRAF v600e mutation in papillary thyroid carcinoma. , 2013, Surgery.

[4]  A. Chou,et al.  BRAFV600E immunohistochemistry in conjunction with mismatch repair status predicts survival in patients with colorectal cancer , 2013, Modern Pathology.

[5]  Aung Ko Win,et al.  BRAFV600E Immunohistochemistry Facilitates Universal Screening of Colorectal Cancers for Lynch Syndrome , 2013, The American journal of surgical pathology.

[6]  A. Voigt,et al.  BRAF V600E‐specific immunohistochemistry for the exclusion of Lynch syndrome in MSI‐H colorectal cancer , 2013, International journal of cancer.

[7]  A. Ristimäki,et al.  BRAF mutation in sporadic colorectal cancer and Lynch syndrome , 2013, Virchows Archiv.

[8]  M. Bronner,et al.  BRAF V600E mutation detection by immunohistochemistry in colorectal carcinoma , 2013, Genes, chromosomes & cancer.

[9]  M. Zhu,et al.  Predictive and prognostic roles of BRAF mutation in patients with metastatic colorectal cancer treated with anti‐epidermal growth factor receptor monoclonal antibodies: A meta‐analysis , 2013, Journal of digestive diseases.

[10]  T. Grogan,et al.  Mutation‐specific antibody detects mutant BRAFV600E protein expression in human colon carcinomas , 2013, Cancer.

[11]  Kyoung-Mee Kim,et al.  Comparison of Three BRAF Mutation Tests in Formalin-Fixed Paraffin Embedded Clinical Samples , 2013, Korean journal of pathology.

[12]  J. Hornick,et al.  Immunohistochemistry using the BRAF V600E mutation‐specific monoclonal antibody VE1 is not a useful surrogate for genotyping in colorectal adenocarcinoma , 2013, Histopathology.

[13]  Ali Salajegheh,et al.  Clinicopathological relevance of BRAF mutations in human cancer , 2013, Pathology.

[14]  J. Honegger,et al.  VE1 immunohistochemistry in pituitary adenomas is not associated with BRAF V600E mutation , 2013, Acta Neuropathologica.

[15]  P. Ladenson,et al.  Association between BRAF V600E mutation and mortality in patients with papillary thyroid cancer. , 2013, JAMA.

[16]  Alexia Iasonos,et al.  BRAF Mutation is associated with early stage disease and improved outcome in patients with low‐grade serous ovarian cancer , 2013, Cancer.

[17]  A. von Deimling,et al.  Immunohistochemistry Is Highly Sensitive and Specific for the Detection of V600E BRAF Mutation in Melanoma , 2013, The American journal of surgical pathology.

[18]  A. von Deimling,et al.  Utilization of a MAB for BRAF(V600E) detection in papillary thyroid carcinoma. , 2012, Endocrine-related cancer.

[19]  A. Pinchera,et al.  The BRAF(V600E) mutation is an independent, poor prognostic factor for the outcome of patients with low-risk intrathyroid papillary thyroid carcinoma: single-institution results from a large cohort study. , 2012, The Journal of clinical endocrinology and metabolism.

[20]  M. Copin,et al.  High prevalence of BRAF V600E mutations in Erdheim-Chester disease but not in other non-Langerhans cell histiocytoses. , 2012, Blood.

[21]  P. Gibbs,et al.  A Highly Sensitive Immunohistochemical Assay to Detect Braf V600E Mutations in Patients with Colorectal Cancer , 2012 .

[22]  A. Hauschild,et al.  Dabrafenib in BRAF-mutated metastatic melanoma: a multicentre, open-label, phase 3 randomised controlled trial , 2012, The Lancet.

[23]  A. von Deimling,et al.  Immunohistochemical Detection of the BRAF V600E-mutated Protein in Papillary Thyroid Carcinoma , 2012, The American journal of surgical pathology.

[24]  James Ziai,et al.  BRAF mutation testing in clinical practice , 2012, Expert review of molecular diagnostics.

[25]  K. Bloom,et al.  Multisite analytic performance studies of a real-time polymerase chain reaction assay for the detection of BRAF V600E mutations in formalin-fixed, paraffin-embedded tissue specimens of malignant melanoma. , 2012, Archives of pathology & laboratory medicine.

[26]  J. Pichler,et al.  Immunohistochemical testing of BRAF V600E status in 1,120 tumor tissue samples of patients with brain metastases , 2012, Acta Neuropathologica.

[27]  A. Hauschild,et al.  Improved survival with vemurafenib in melanoma with BRAF V600E mutation. , 2011, The New England journal of medicine.

[28]  S. Pileri,et al.  BRAF mutations in hairy-cell leukemia. , 2011, The New England journal of medicine.

[29]  H. Zentgraf,et al.  Assessment of BRAF V600E mutation status by immunohistochemistry with a mutation-specific monoclonal antibody , 2011, Acta Neuropathologica.

[30]  R. Jorissen,et al.  Optimizing targeted therapeutic development: Analysis of a colorectal cancer patient population with the BRAFV600E mutation , 2011, International journal of cancer.

[31]  G. Mann,et al.  Prognostic and clinicopathologic associations of oncogenic BRAF in metastatic melanoma. , 2011, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[32]  A. Iafrate,et al.  BRAF V600E Mutations Are Common in Pleomorphic Xanthoastrocytoma: Diagnostic and Therapeutic Implications , 2011, PloS one.

[33]  J. O’Leary,et al.  BRAFV600E: Implications for Carcinogenesis and Molecular Therapy , 2011, Molecular Cancer Therapeutics.

[34]  S. Ariyan,et al.  Incidence of the V600K mutation among melanoma patients with BRAF mutations, and potential therapeutic response to the specific BRAF inhibitor PLX4032 , 2010, Journal of Translational Medicine.

[35]  A. Nicholson,et al.  Mutations of the BRAF gene in human cancer , 2002, Nature.

[36]  C. Toon,et al.  BRAF V600E mutation specific immunohistochemistry with clone VE1 is not reliable in pituitary adenomas. , 2014, Pathology.

[37]  D. Capper,et al.  Improved molecular classification of serrated lesions of the colon by immunohistochemical detection of BRAF V600E , 2014, Modern Pathology.

[38]  M. Zeiger,et al.  BRAF V600E Mutation Independently Predicts Central Compartment Lymph Node Metastasis in Patients with Papillary Thyroid Cancer , 2012, Annals of Surgical Oncology.