An Evaluation on the Potential Clinical Application of PCR-SSCP Screening EGFR Mutations in Non-small Cell Lung Cancer
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摘要:
目的 评价PCR-SSCP筛检非小细胞肺癌(NSCLC)ECFR基因突变的临床应用潜力。 方法 分别采用DNA测序法和PCR-SSCP分析对一定数量的NSCLC标本进行EGFR基因突变检测, 以DNA测序结果为金标准, 计算PCR-SSCP法的灵敏度、假阴性率、特异度、假阳性率、约登指数、粗符合率、预测值和似然比等指标; 同时随机抽取20%的样本, 重新进行PCR-SSCP分析, 计算两次结果的Kappa指数值。 结果 PCR-SSCP分析的灵敏度为97.2%, 假阴性率为2.8%, 特异度为94.3%, 假阳性率为5.7%, 约登指数为0.915, 粗符合率为94.8%, 阳性预测值为77.8%, 阴性预测值为99.4%, 阳性似然比为17.1, 阴性似然比为0.5, 前后两次PCR-SSCP分析的Kappa指数值为0.81(P < 0.05)。 结论 PCR-SSCP检测NSCLC EGFR基因突变具有较高的真实性、可靠性和实用性。 Abstract:Objective To evaluate the clinical application potentiality of PCR-single strand conformation polymorphism (PCR-SSCP)used to screen EGFR mutations in non-small cell lung cancer(NSCLC). Methods Certain NSCLC specimens were tested with PCR-SSCP analysis and DNA sequencing respectively, and then some related indices were calculated, such as sensitivity, false negative rate, specificity, false positive rate, Youden index, crude agreement rate, likelihood ratio, and predictive value, taking the results of DNA sequencing as the "gold standard".At the same time, 20%of the tested specimens were randomly sampled for a second PCR-SSCP analysis, and the value of Kappa index was calculated by comparing the results from two PCR-SSCP analyses. Results Of PCR-SSCP analysis, sensitivity was 97.2%, false negative rate was 2.8%, specificity was 94.3%, false positive rate was 5.7%, Youden index was 0.915, crude agreement rate was 94.8%, positive likelihood ratio was 17.1, negative likelihood ratio was 0.5, positive predictive value was 77.8%, negative predictive value was 99.4%, and Kappa index was 0.81(P < 0.05). Conclusion PCR-SSCP analysis has high validity, reliability and practicability in screening EGFR mutations in NSCLC. -
图 1 EGFR基因exon19、exon21 PCR产物的琼脂糖凝胶电泳
M：DNA分子量标准；1：外显子19；2：外显子21
Figure 1. Electrophotogram of the target PCR product of EGFR exon19 and exon21
表 1 NSCLC EGFR基因外显子19和21的突变类型分布
Table 1. The distribution of mutation types in EGFR exon 19 and exon 21 in NSCLC
表 2 PCR-SSCP检测NSCLC EGFR基因突变与DNA测序结果的比较 例
Table 2. The comparison of PCR-SSCP detecting EGFR mutation with DNA sequencing
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