癌变·畸变·突变 ›› 2010, Vol. 22 ›› Issue (6): 444-447.doi: 10.3969/j.issn.1004-616x.2010.06.008

• 论著 • 上一篇    下一篇

高分辨率熔解曲线分析技术在检测非小细胞肺癌组织EGFR基因突变中的应用

由　田1，2刘　政2，邱广斌2,   

  1. 1. 中国医科大学第一临床学院急诊科，辽宁 沈阳 110001； 2. 解放军二零二医院检验科，辽宁 沈阳 110003
  • 收稿日期:2010-07-15 修回日期:2010-09-25 出版日期:2010-11-30 发布日期:2010-11-30
  • 通讯作者: 邱广斌

Detecting EGFR mutations in non_small cell lung carcinoma patients using high resolution melting curve analysis

YOU Tian1，2, LIU Zheng2, QIU Guang-bin2，   

  1. 1. Department of Emergency Medicine, the First Clinical School of China Medical University, Shenyang 110001; 2.Department of Laboratory Medicine,No.202 Hospital of PLA, Shenyang 110003,Liaoning, China
  • Received:2010-07-15 Revised:2010-09-25 Online:2010-11-30 Published:2010-11-30
  • Contact: QIU Guang-bin

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摘要： 目的： 探索高分辨率熔解曲线(high resolution melting, HRM)分析技术在检测肺癌组织表皮生长因子受体(epidermal grouth factor receptor, EGFR)基因突变中的应用。 方法： 利用HRM技术，进行人肺癌组织EGFR基因外显子19和21突变筛查，并做测序验证。 结果： 外显子19的HRM分型结果显示，52例样品中有3种类型熔解曲线，45例样品为A型，5例为B型，2例为C型。外显子21的分析结果显示有2种熔解曲线，49例样品为N型，3例样品为M型。测序结果显示，外显子19 A型熔解曲线代表外显子19的野生型，B型熔解曲线代表杂合性缺失delE746_A750(c.2235_2249del)，C型熔解曲线代表杂合性缺失delL747_S752(c.2240_2257del)。外显子21 N型熔解曲线代表野生型，M型熔解曲线代表杂合性L858R(c.2573T_G)，HRM分析结果与测序结果完全一致。 结论： 本研究表明HRM分析技术能准确检测EGFR基因突变，是一种适合肺癌组织中EGFR基因突变筛查的方法

关键词: 非小细胞肺癌, 表皮生长因子受体, 高分辨率熔解曲线

Abstract: OBJECTIVE: To evaluate the feasibility and sensitivity of detecting the epidermal growth factor receptor(EGFR) mutation in non_small cell lung carcinoma(NSCLC) patients using high resolution melting(HRM) curve analysis. METHODS: HRM was applied to detect the mutations of EGFR exon 19 and 21 in NSCLC patients, the results were then confirmed by direct sequencing. RESULTS: Three melting curves were obtained by HRM anlysis for EGFR exon 19 and two melting curves for exon 21. Type A(45/52), B(5/52), C(2/52) melting curves represent wild type , heterozygous delE746_A750 and delL747_S752 of exon 19, respectively.Type N(49/52) and M(3/52) melting curves represent wild type and heterozygous L858R of exon 21. Results of HRM were consistent with those of direct sequenc. CONCLUSION: HRM is an accurate technique to detect EGFR mutations. It is also a low_cost test which is suitable for screening.

Key words: non_small cell lung carcinoma, epidermal growth factor receptor, high resolution melting curve

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