直接测序法与荧光定量PCR法检测非小细胞肺癌EGFR基因突变对比分析

摘要

目的：比较直接测序和荧光定量PCR法检测非小细胞肺癌（ NSCLC） EGFR基因突变状态结果的差异。方法：分别采用荧光定量PCR（蝎形探针扩增阻滞突变系统ARMS）和直接测序法检测155例石蜡包埋NSCLC样本中EGFR基因18，19，20，21号外显子突变状态，比较检测结果的差异。结果：80例手术切除样本中，直接测序检测EGFR突变阳性者25例（31．25％）， ARMS法为28例（35％），差异无统计学意义（P＞0．05）；75例活检样本中，直接测序检测EGFR突变阳性者19例（25．33％）， ARMS法为26例（34．67％），差异具有统计学意义（P＜0．05）。结论：对于手术切除样本，直接测序和ARMS法均可有效检测出EGFR基因突变状态；但对于活检组织样本，ARMS较直接测序灵敏性更高。%Objective:To compare the differences between fluorescence quantitative PCR and direct sequencing of detecting EGFR mutations in non-small cell lung cancer.Methods:Both direct sequencing and fluorescence quantitative PCR ( amplification refractory mutation system, ARMS) were used to detect EGFR mutations in exon 18, 19, 20, and 21 in 155 formalin-fixed and parrffin-embeded ( FFPE) NSCLC specimens,and the differences between the two methods were analyzed.Results:In 80 surgical excision samples, EG-FR mutations were identified in 25 cases with a mutation rate of 31.5%by direct sequencing, and in 28 cases with a mutation rate of 35%by ARMS, showing no statistical differences in the two detections(P>0.05).In 75 biopsy specimens, EGFR mutations were i-dentified in 19 cases with a mutation rate of 25.33% by direct sequencing, while in 26 cases with a mutation rate of 34.67% by ARMS.There were significant differences of EGFR mutation rate between the two methods(P<0.05).Conclusion:Both direct se-quencing and ARMS can effectively identify EGFR mutation in surgical excision samples, but in biopsy specimens, ARMS gains a higher sensitivity than direct sequencing in detection of EGFR mutation.