突变体富集PCR法检测非小细胞肺癌病理组织EGFR基因突变

Detection of EGFR Mutations in Non-Small Cell Lung Cancer by Mutant-Enriched PCR

目的建立突变体富集PCR法榆测非小细胞肺癌病理组织中的EGFR基因突变。方法选取55例细支气管肺泡癌和59例非小细胞肺癌病理组织蜡块，提取基因组DNA，采用不同的突变体富集PCR法（PCR—PAGE和PCR—RLFP）检测EGFR基因常见的19和21外显子突变，并经过直接测序验证。结果在59例非小细胞肺癌中共检测出EGFR基因突变22例，突变率为37．3％（22／59）。55例细支气管肺泡癌中共检测出24例基因突变，突变率为43．6％（24／55）。经直接测序验证，EGFR19外显予有3种类型缺失突变。EGFR21外显子的错义突变为L858R。结论突变体富集PCR法准确、快速、经济，便于临床筛查非小细胞肺癌病理组织中的EGFR基因突变。

Objective To detect EGFR mutations in non-small-cell lung cancer （NSCLC） by mutant-enriched PCR. Methods Paraffinembedded tumor tissue samples were obtained from 55 patients with bronchioloalveolar carcinoma and 59 patients with advanced NSCLC for the extraction of genomic DNA. The deletion mutation in EGFR exon 19 was detected by PCR-PAGE and the point mutation in EGFR exon 21 was detected by PCR-RLFP. The mutations were all subjected to verification by direct sequencing. Results 22 EGFR mutations from 59 NSCLC samples （22/59,37.3 %） and 24 mutations from 55 bronchioloalveolar carcinoma samples （24/55,43.6 %） were detected. Three kinds of EGFR deletion mutation in exon 19 and missense mutation L858R in exon 21 were verified by direct sequencing. Conclusion The mutantenriched PCR was accurate, rapid, economical, and convenient for the screening of EGFR mutations in NSCLC.