【摘要】 目的探讨应用双标准曲线的实时荧光PCR法检测原癌基因人类表皮生长因子受体2(HER2)基因扩增在临床乳腺癌诊治中的可行性。方法收集500例乳腺癌术后新鲜组织标本,抽提组织DNA进行实时荧光PCR检测,采用双标准曲线法定量,通过计算目的基因浓度和内标基因浓度的比值来判断HER2基因的扩增情况。选择灵敏度和特异度均较高的荧光原位杂交方法作为对照方法。结果检测阳性标本72例,阴性样本419例。该方法检测的灵敏度为85.9%,特异度为98.79%,准确度为96.74。与荧光原位杂交法(FISH)检测结果相比,二者具有较好的一致性。结论双标准曲线的实时荧光PCR法用于检测HER2基因扩增相对准确可靠,有较好的临床应用前景。更多还原

【Abstract】 Objective To evaluate the double standard curve of Real-time fluorescence PCR to detect human epidermalgrowth factor receptor-2(HER2) gene amplification for breast cancer diagnosis and treatment.Methods Genomic DNA was extracted in 500 cases fresh tissue samples of breast cancer for real-time fluorescence PCR and quantified by double standard curve.Data was analyzed by calculating the concentration ratio of the target gene to the referenced gene detected.Fluorescent in situ hybridization(FISH) was chosen as the control method for HER2 detection.Results 72 cases of positive samples and 419 negative samples were detected by this method.The sensitivity was 85.9%,the specificity was 98.79% and the accuracy is 96.74%.Compared with the FISH test,the results of both methods have good consistency.Conclusion The method of double standard curve of Real-time fluorescence PCR to detect HER2 gene amplification has a wide application prospect for breast cancer diagnosis and treatment.更多还原