乳腺癌组织HER2基因扩增的检测方法

摘要

目的:评价高分辨率熔体聚合酶链反应(high-resolution melt polymerase chain reaction,HRM-PCR)检测HER2基因扩增的有效性及其与荧光原位杂交(fluorescence in situ hybridization,FISH)和免疫组织化学(immunohistochemistry,IHC)检测法的一致性.方法:采用HRM-PCR法检测HER2阴性及阳性细胞株,评估检测方法的有效性;检测98例已行FISH和/或IHC的临床样本,并与FISH和IHC结果进行比较.结果:HRM-PCR检测可以有效区分HER2阴性及阳性细胞株(P<0.05),具有较好的可重复性;检测98例临床样本显示,阴性和阳性样本检测结果差异有统计学意义(0.18±0.14 vs 1.42±0.88,P<0.01),与IHC的一致性为80.33%(kappa=0.6,P<0.01),与FISH的一致性为87.88%(kappa=0.7,P<0.01),结论:HRM-PCR是一种可靠有效的检测HER2基因扩增的方法,与FISH和IHC均有较好的一致性.%Objective: To evaluate the detection of HER2 gene amplification effectiveness by high-resolution melt polymerase chain reaction (HRM-PCR) and compare with fluorescence in situ hybridization (FISH) and/ or immunohistochemistry (IHC). Methods: HRM-PCR was used to test 98 clinical samples which has detected by FISH and/or IHC, and the results of FISH and IHC were compared. Results: HER2 negative and positive cell lines can be distinguished by HRM-PCR (P<0.05) with well repeatability. The difference between negative and positive samples results was significant (0.18±0.14 vs 1.42±0.88, P<0.01),and 80.33% consistency with IHC (kappa=0.6, P<0.01), 87.88% with FISH (kappa=0.7, P<0.01).Conclusion: HRM-PCR is a reliable and effective method to detect HER2 gene amplification which has a high consistency with FISH/IHC.