摘要
目的 比较微滴数字PCR（ddPCR）与突变扩增阻滞系统（ARMS）技术在检测非小细胞肺癌不同类型标本表皮生长因子受体（EGFR）基因突变的差异,并探讨ddPCR在临床上的应用价值.方法 收集2016年79例非小细胞肺癌患者标本,其中包括胸水细胞块22例,手术标本18例,活检穿刺标本12例,血浆标本27例,并经ARMS法检测已知其EGFR基因突变情况,现用ddPCR法检测上述标本EGFR基因突变情况.结果 79例标本中,18例手术标本和12例活检穿刺标本ddPCR法检测与ARMS法检测结果完全一致,阳性比例分别为9/18和5/12;22例胸水细胞块中ARMS法阳性检出率为40.9%（9/22）,ddPCR法阳性检出率为45.5%（10/22）,其中有2例ddPCR法分别检测出EGFR基因19-del＋T790M双突变和L858R＋T790M双突变,而ARMS法检测分别为EGFR基因19-del和L858R,还有1例ddPCR检测为L858R突变,而ARMS法检测为阴性,其余19例结果一致;27例血浆标本中ARMS法阳性检出率为33.3%（9/27）,ddPCR法阳性检出率为37.0%（10/27）,其中2例ddPCR检测分别为L858R突变和19-del＋T790M双突变,ARMS法检测仅为阴性和19-del,其余25例结果一致.结论 对于非小细胞肺癌患者胸水细胞块和血浆标本EGFR检测,ddPCR法较ARMS法具有更高的灵敏度,在临床上具有更广的应用价值.
Objective To compare amplification refractory mutation system（ARMS）and droplet digital PCR（ddPCR）in the detection of epidermal growth factor receptor（EGFR）gene mutations in patients with non-small cell lung cancer（NSCLC）,and to investigate the clinical value of ddPCR. Methods A total of 79 specimens of NSCLC, including 22 cases of cell block, 18 cases of surgical specimens, 12 cases of biopsy specimens and 27 cases of plasma samples,were analyzed for the mutation status of EGFR gene by ARMS and droplet digital PCR method. Results In 18 cases of surgical specimens and 12 cases of biopsy specimens, the detection results by the two methods were identical with positive rates of 9/18 and 5/12, respectively. In 22 cases of effusion cell blocks,ARMS detected 19-del and L858R of EGFR gene in two cases,in which droplet digital PCR detected 19-del＋T790M mutations in one case and L858R＋T790M mutation in another. L858R mutation was detected by droplet digital PCR in one case but ARMS assay was negative. The remaining 19 cases were consistent by the two methods. In blood samples, the positive rate was 33.3%（9/27）by ARMS and 37.0%（10/27）by droplet digital PCR. Two cases showed L858R and 19-del＋T790M mutation by droplet digital PCR but ARMS assay detected only 19-del. The remaining 25 cases were consistent by the two methods. Conclusion Droplet digital PCR method is more sensitive and accurate than ARMS for the detection of EGFR mutations in pleural fluid and blood samples, can be used in clinical test.
出处
《中华病理学杂志》
CAS
CSCD
北大核心
2017年第11期764-768,共5页
Chinese Journal of Pathology
关键词
癌
非小细胞肺
受体
表皮生长因子
核酸扩增技术
Carcinoma
non-small-cell lung
Receptor
epidermal growth factor
Nucleic acid amplification techniques
