ddPCR与ARMS-PCR检测NSCLC患者EGFR基因T790M突变的比较研究

Comparative Analysis of EGFR T790M Detection by ddPCR and ARMS-PCR in NSCLC Patients

目的:通过对比微滴式数字PCR(droplet digital PCR,ddPCR)和突变扩增阻滞系统PCR(amplification refractory mutation system PCR,ARMS-PCR)检测非小细胞肺癌(non-small cell lung cancer,NSCLC)表皮生长因子受体(epidermal growth factor receptor,EGFR)基因T790M突变的检测效能,综合分析两种检测方法的优势及适用范围。方法:回顾性收集115例NSCLC患者的标本,同时使用ddPCR和ARMS-PCR两种方法检测EGFR基因T790M突变状态,比较两种方法的检测结果,并分析不同样本类型对ddPCR检测结果的影响。结果:ddPCR检测T790M阳性率为65.2%,ARMS-PCR阳性率为27.8%,ddPCR检测敏感性显著高于ARMS-PCR(P<0.05),两种方法检测一致率为62.6%。ddPCR+ARMS-样本的T790M平均阳性微滴数和平均突变丰度(11,0.35%)均显著低于ddPCR+ARMS+样本(632.3,17.7%)。ddPCR检测基因组DNA和外周血游离DNA阳性率分别为55.9%和78.7%,不同样本类型T790M突变阳性微滴数和突变丰度无显著差别。结论:ddPCR和ARMS-PCR均可用于NSCLC患者EGFR基因T790M突变检测,二者具有良好的一致性,ddPCR具有更高的灵敏性,在检测突变丰度较低的样本方面具有优势。

Objective:This study aimed to compare the detective efficacy of droplet digital polymerase chain reaction(ddPCR)and amplification refractory mutation system(ARMS)-PCR in detecting epidermal growth factor receptor(EGFR)T790M mutation in non-small cell lung cancer(NSCLC)patients,and evaluate the clinical application value of these two techniques.Methods:115 specimens retrospectively collected from NSCLC patients were simultaneously detected by ddPCR and ARMS-PCR,and the results were comprehensively analyzed.The influence of different sample type on detecting results was also explored.Results:The T790M positive rate by ddPCR and ARMS-PCR were 65.2%and 27.8%,respectively.The sensitivity of ddPCR was significantly higher than that of ARMS-PCR(P<0.05),and the consistency of these two methods was 62.6%.The mean T790M mutant droplet number and mean mutant abundance of ddPCR+ARMS-group(11,0.35%)were significantly lower than those of the ddPCR+ARMS+group(632.3,17.7%).The positive rates of T790M mutation in genomic DNA and peripheral blood cell-free DNA by ddPCR are 55.9%and 78.7%,respectively.There was no significant difference between the mean T790M mutant droplet number and mean mutant abundance in different sample types.Conclusion:ddPCR and ARMS-PCR are both convenient and efficient methods for T790M detection,and they are relatively consistent with each other.However,ddPCR has a higher sensitivity than ARMS-PCR,so that it has an advantage in detecting samples with low mutant abundance.