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  • Title: [Utility and evaluation of immunohistochemical detection of BRAF V600E mutation in melanoma].
    Author: Lyu JJ, Kong YY, Cai X, Shen XX, Lu YW, Ren M.
    Journal: Zhonghua Bing Li Xue Za Zhi; 2017 Aug 08; 46(8):548-552. PubMed ID: 28810295.
    Abstract:
    Objective: To evaluate the sensitivity, specificity and clinical value of anti-BRAF V600E antibody (clone VE1) in detection of the BRAF V600E mutant in formalin-fixed and paraffin-embedded (FFPE) melanoma specimens by immunohistochemical (IHC) methods. Methods: A total of 50 melanoma samples collected between 2008 and 2016 from 40 patients were analyzed for BRAF mutation (exon 15) by DNA sequencing using FFPE. These tissues were immunostained with VE1 antibody, and the results were analyzed and compared with those by DNA sequencing. Results: By DNA sequencing, 36 cases showed BRAF mutation while others were BRAF wild type. Among the 36 cases with BRAF mutation, 32 harbored BRAF V600E, two harbored BRAF V600K, one had BRAF K601E and one had BRAF D594N, respectively. IHC staining showed 30 specimens were VE1 positive, while 19 were negative. The determination of IHC result for one case was obscured by heavy pigments. Of the BRAF-mutated specimens, four specimens with BRAF mutation other than V600E were all negative for VE1. The sensitivity and specificity of the VE1 immunostaining was 96.8% and 100.0% respectively.Concordance of BRAF V600E detection between immunostaining and DNA sequencing was 98.0%(48/49). Conclusions: High sensitivity and specificity for VE1 immunostaining in detecting BRAF V600E in melanomas are demonstrated. It is a rapid and cost-effective method for detecting BRAF V600E mutations in melanoma patients. Hence, VE1 immunostaining can be used as an important screening method for BRAF mutation in laboratories. 目的： 探讨恶性黑色素瘤患者石蜡标本中用免疫组织化学抗BRAF V600E抗体检测BRAF V600E突变的临床应用价值，分析免疫组织化学法检测BRAF V600E的灵敏度和特异度。 方法： 应用PCR扩增和直接测序方法检测2008至2016年间40例恶性黑色素瘤患者的50份石蜡包埋组织标本中BRAF第15号外显子突变情况，并应用免疫组织化学法检测BRAF V600E抗体表达情况，比较免疫组织化学与基因检测结果。 结果： BRAF突变分子检测阳性样本36份，其中BRAF V600E突变32份，BRAF V600K突变2份，BRAF K601E和BRAF D594N突变各1份，其余14份均为BRAF野生型。BRAF V600E抗体免疫组织化学检测阳性样本30份，阴性19份，1份因色素过多无法判断免疫组织化学。4份具有BRAF其他类型突变的样本用免疫组织化学方法检测均为阴性。BRAF V600E抗体免疫组织化学检测敏感性为96.8%，特异性为100.0%，免疫组织化学检测与分子检测总符合率为98.0%(48/49)。 结论： 在恶性黑色素瘤中使用免疫组织化学法检测BRAF V600E突变蛋白表达情况，其敏感性及特异性均较高，简便、快速、价格相对低廉，可以作为临床上检测恶性黑色素瘤BRAF突变的重要筛查方法。.
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