摘要
目的通过对比二代测序(NGS)和突变阻滞扩增系统PCR(ARMS-PCR)对肺癌靶向基因突变的检出率,阐释其检测特点和应用优势。方法选取2018年1月至2020年3月从河南省内各地区医疗机构送至郑州金域临床检验中心有限公司检测肺癌靶向基因的病例。筛选出1046例样本,临床诊断结果均为疑似或确诊肺癌。样本包括石蜡包埋组织(FFPE)772例、新鲜血浆274例。在FFPE样本中,采用NGS技术检测的有329例,采用ARMS-PCR技术检测的有443例。在新鲜游离血浆样本中,采用NGS技术检测的有83例,采用ARMS-PCR技术检测的有191例。分别比较NGS技术与ARMS-PCR技术对不同样本肺癌靶向基因(包括EGFR、KRAS、PIK3CA、TP53、BRAF等22个基因)的检出率。结果NGS技术与ARMS-PCR技术对FFPE样本EGFR基因已知位点突变总检出率比较,差异无统计学意义(P>0.05)。ARMS-PCR技术对FFPE样本外显子21(L858R)突变的检出率高于NGS技术(P<0.05)。NGS技术与ARMS-PCR技术对血浆样本EGFR基因主要位点突变外显子20(T790M)、外显子19(缺失突变)、外显子21(L858R)突变总检出率比较,差异无统计学意义(P>0.05)。NGS技术检出EGFR基因扩增16例,检出率为4.86%(16/329)。NGS检测肺癌靶向基因套餐的FFPE样本中,214例确诊为肺腺癌,34例确诊为肺鳞癌。与肺腺癌比较,NGS技术对FFPE样本肺鳞癌TP53和PIK3CA基因突变检出率较高,对EGFR基因突变检出率较低(P<0.05)。结论ARMS-PCR技术对EGFR基因已知位点突变总检出率与NGS接近,但对某些单个已知突变位点检出率与NGS相比可能略高;NGS靶向基因检测技术通量高,可检测未知突变及多种突变;两种方法在肺癌靶向治疗中各有优势,临床应合理搭配应用和选择。
Objective To compare the detection rate of target gene mutation in lung cancer between next generation sequencing(NGS)and amplification refractory mutation system-PCR(ARMS-PCR),and explain detection characteristics and application advantages.Methods The cases of lung cancer targeting genes were selected from medical institutions in Henan Province to Zhengzhou KingMed Center for Clinical Laboratory Co.,Ltd.from January 2018 to March 2020.The 1046 cases in the clinical diagnosis of suspected or diagnosed lung cancer were screened.There were 772 cases of formalin fixed paraffin-embedded(FFPE)and 274 cases of fresh plasma in the samples.In the FFPE samples,329 cases were detected by NGS technology and 443 cases were detected by ARMS-PCR technique.In the fresh plasma samples,83 cases were detected by NGS technology and 191 cases were detected by ARMS-PCR technique.The detection rates of different lung cancer target genes(including 22 genes such as EGFR、KRAS、PIK3CA、TP53、BRAF)between NGS technique and ARM-PCR technique were compared respectively.Results There was no difference in the total detection rate of mutations at known EGFR gene sites of FFPE samples between NGS technique and ARMS-PCR technique(P>0.05).The detection rate of exon 21(L858R)mutation in FFPE samples in ARMS-PCR technique was higher than that in NGS technique(P<0.05).There were no differences in the positive detection rate of mutation at main EGFR gene sites such as exon 20(T790M),exon 19(deletion mutation),exon 21(L858R)mutations between NGS technique and ARMS-PCR technique(P>0.05).There were 16 cases of EGFR gene amplification detected by NGS,with a detection rate of 4.86%(16/329).In the FFPE samples of NGS targeted gene package for lung cancer,214 cases were diagnosed as lung adenocarcinoma and 34 cases as lung squamous cell carcinoma.The detection rate of TP53 and PIK3CA gene mutations in FFPE lung squamous cell carcinoma was higher than that in lung adenocarcinoma(P<0.05).Conclusion The total detection rate of known gene mutations in NGS could be close to that of ARMS-PCR.The detection rate of some known mutation sites by ARMS-PCR may be slightly higher than that of NGS.NGS targeted gene detection has higher throughput and wider range,which can detect unknown mutations and a larger variety of mutations.The two technologies have different advantages in the targeted therapy for lung cancer,and should be chosen based on the clinical question.
作者
朱丽蒙
马楠
任晓妮
张暋
ZHU Li-meng;MA Na;REN Xiao-ni;ZHANG Min(Laboratory diagnostics,Zhengzhou KingMed Center for Clinical Laboratory Co.,Ltd.,Zhengzhou 450000,China)
出处
《河南医学研究》
CAS
2020年第33期6152-6156,共5页
Henan Medical Research
关键词
肺癌
靶向基因
二代测序
突变阻滞扩增系统PCR
lung cancer
targeted gene
next generation sequencing
refractory mutation system-PCR
