CAJ | 학술논문

目的探讨特异引物双环探针扩增实时PCR技术表皮生长因子受体(EGFR)检测方法(ADx-EGFR实时PCR法)和Sanger DNA测序法在肺癌EGFR基因体细胞突变检测中的临床价值.方法收集肺癌组织石蜡切片208例,分别采用ADx-EGFR实时PCR法和Sanger DNA测序法检测肺癌组织中EGFR基因外显子18、19、20、21的突变类型,计算其突变率,分析两种检测方法检测EGFR基因突变的一致性.结果 208例肺癌组织中,ADx-EGFR实时PCR法成功检测208例,检出EGFR基因突变40例,突变检出率为19.2％；Sanger DNA测序法成功检测196例,检出突变22例,突变检出率为11.2％.肺癌组织中主要以外显子19缺失和外显子21上的L858R的点突变为主,分别占4.8％ (10/208)和11.6％ (23/208),其余的突变类型少见.结论对于甲醛固定石蜡包埋组织而言,ADx-EGFR实时PCR法检测EGFR基因成功率和突变检出率均高于Sanger DNA测序法,可成为临床上检测肿瘤EGFR基因突变的方法.
목적 탐토특이인물쌍배탐침확증실시PCR기술표피생장인자수체(EGFR)검측방법(ADx-EGFR실시PCR법)화Sanger DNA측서법재폐암EGFR기인체세포돌변검측중적림상개치.방법 수집폐암조직석사절편208례,분별채용ADx-EGFR실시PCR법화Sanger DNA측서법검측폐암조직중EGFR기인외현자18、19、20、21적돌변류형,계산기돌변솔,분석량충검측방법검측EGFR기인돌변적일치성.결과 208례폐암조직중,ADx-EGFR실시PCR법성공검측208례,검출EGFR기인돌변40례,돌변검출솔위19.2％；Sanger DNA측서법성공검측196례,검출돌변22례,돌변검출솔위11.2％.폐암조직중주요이외현자19결실화외현자21상적L858R적점돌변위주,분별점4.8％ (10/208)화11.6％ (23/208),기여적돌변류형소견.결론 대우갑철고정석사포매조직이언,ADx-EGFR실시PCR법검측EGFR기인성공솔화돌변검출솔균고우Sanger DNA측서법,가성위림상상검측종류EGFR기인돌변적방법.
Objective To map the frequency and types of EGFR gene mutations present in lung cancer tissues.To evaluate the clinical applicability of a novel real-time double-loop probe PCR of which the ADx-EGFR kit is based,and to compare its performance with traditional Sanger DNA sequencing in the detection of somatic mutations of tumor genes.Methods A total of 208 formalin-fixed paraffin-embedded (FFPE) tumor samples were tested.Genomic DNA of the tissue samples was extracted and purified,and subjected to both traditional PCR amplification,Sanger sequencing of EGFR gene in exon 18,19,20,21,and ADx's EGFR mutation detection kit.The mutation rates for EGFR gene in exon 18,19,20,21,as well as the frequency of each mutation detected by the two methods,were analyzed.Results The traditional Sanger DNA sequencing technique was successfully performed in 196 out of 208 (94.2％) lung cancer samples,and 22 samples (11.2％) showed EGFR gene mutations.ADx-EGFR kit was successfully used in the lung cancers of all of the 208 cases (100.0％),and 40 samples (19.2％) showed mutations.In the lung cancer samples analyzed,mutations were mainly detected in the exon 19 and exon 21 L858R point mutation,i.e.4.8％ (10/208) and 11.6％ (23/208) of total mutations,respectively,and the remaining mutations were rare.Conclusions The success rate of ADx-EGFR real-time PCR for formalin-fixed and paraffin-embedded tissues samples is significantly higher than that of Sanger sequencing (P ＜0.01).There are significant differences between the two methods.ADx-EGFR real-time PCR shows a much higher successful detection rate and mutation rate of lung cancer tissues compared with that of Sanger sequencing.As a result,the real-time PCR with ADx-EGFR kit is proved to have a good clinical applicability and a strong advantage over the traditional Sanger DNA sequencing.It is an effective and reliable tool for clinical screening of somatic gene mutations in tumors.