摘要
目的分析微滴式数字PCR技术(droplet digital PCR,dd PCR)在非小细胞肺癌(non-small-cell lung cancer,NSCLC)患者外周血EGFR基因突变检测中的应用价值。方法采用dd PCR法与扩增阻滞突变系统技术(amplification refractory mutation system,ARMS)分别检测58例NSCLC患者及TKI治疗后耐药患者20例外周血EGFR基因突变,Kappa检验验证dd PCR法检测EGFR基因突变结果与ARMS法的一致性,并对检测结果进行分析比较。结果 ARMS法检测EGFR基因突变共检出34例患者,其中双突变患者5例,dd PCR法共检出38例患者,其中双突变患者10例;两者Kappa值=0.72,阳性一致率为97.50%,阴性一致率为72.73%,总一致率为86.30%;dd PCR法检测接受TKI治疗患者的20外显子p.T790M位点突变检出率为50.00%(10/20),而无TKI用药史患者均未检出;dd PCR法检出突变丰度<1%的突变位点占总突变位点的29.17%(14/48),ARMS法仅检出其中的35.71%(5/14)。结论 dd PCR法与ARMS法相比具有更高敏感性,且可绝对定量,EGFR基因突变的检出率更高。
Objective To investigate the application value of droplet digital PCR （ddPCR） in the detection of peripheral blood EGFR mutations of the patients with non-small cell lung cancer （NSCLC）. Methods The EGFR mutations in the peripheral blood of 58 NSCLC patients, including 20 patients resistant to TKI treatment, were detected by ddPCR and the amplification refractory mutation system （ARMS）, respectively. Then, the obtained results were compared, and the consistency between them was analyzed with Kappa test. Results A total of 34 patients with EGFR mutations were detected by ARMS, of which 5 were double mutations. However, 38 patients with EGFR mutations were detected by ddPCR, of which 10 were double mutations. The Kappa value, positive consistency, negative consistency and total consistency between ARMS and ddPCR were 0.72, 97.50%, 72.73% and 86.30%, respectively. The de- tection rate of p.T790M mutation in exon 20 in the patients with TKI treatment by ddPCR was 50% （10/20） , while zero in the patients with no TKI treatment. The mutation sites with the abundance below 1% detected by ddPCR accounted for 29.17% （14/48） of the total mutation sites. However, only 35.71% of them （5/14） were detected by ARMS. Conclusion Compared with ARMS, ddPCR has higher sensitivity and detectable rate of EGFR mutations, and the mutations can be quantified absolutely.
作者
毛旭华
汤俊明
马芬芬
王彦
MAO Xuhua;TANG Junming;MA Fenfen;WANG Yan(Department of Clinical Laboratory,Affiliated Yixing People＇s Hospital of Jiangsu University,Wuxi 214200,Jiangsu;Department of Clinical Laboratory,Lianyungang Second People＇s Hospital,Lianyungang 222000,Jiangsu,China)
出处
《临床检验杂志》
CAS
CSCD
2018年第8期589-592,共4页
Chinese Journal of Clinical Laboratory Science
基金
连云港市医学检验科重点学科建设项目(SH1526)
