Rapid detection of epidermal growth factor receptor mutations in lung cancer by the SMart-Amplification process
Rapid detection of epidermal growth factor receptor mutations in lung cancer by the SMart-Amplification process
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DOI：
10.1158/1078-0432.ccr-07-0509
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发表时间：
2007-09-01
影响因子：
11.5
通讯作者：
Shimada, Hiroshi
中科院分区：
文献类型：
--
作者：
Hoshi, Kanako;Takakura, Hideki;Shimada, Hiroshi
目的：吉非替尼治疗非小细胞肺癌（NSCLC）的阳性反应与表皮生长因子受体（EGFR）基因突变相关。以前的报告主要基于通过测序进行的诊断筛查。然而，测序是一个耗时和复杂的过程，不适合常规临床use.Experimental设计：我们已经开发了快速，简单，灵敏的突变检测分析的基础上的SMART扩增过程（SMAP）和应用它分析EGFR基因突变的临床样本。通过使用SMAP，我们可以在30分钟内检测突变，包括样品制备。为了验证该检测系统在临床诊断中的潜在用途，我们研究了45例NSCLC患者的EGFR突变使用测序和SMAP.Results：的SMAP检测结果完全匹配的测序结果，除了在一种情况下，SMAP是能够确定一个突变，没有检测到测序。我们还评估了SMAP在EGFR突变检测中的敏感性和特异性。在连续稀释研究中，SMAP能够在野生型基因组DNA混合物中仅含有0.1%突变等位基因的样品中发现突变。我们也可以显示扩增突变的DNA，只有30个拷贝，每个reaction.Conclusions：SMAP方法提供了更高的灵敏度和特异性比替代技术，同时消除了测序的需要，以确定突变的EGFR基因的NSCLC。它提供了一个强大的和即时的护理方法，用于快速识别大多数可能对吉非替尼有反应的患者。
Purpose: A positive response to gefitinib in non -small cell lung, cancer (NSCLC) has been correlated to mutations in epidermal growth factor receptor (EGFR) gene. Previous reports have been based mainly on diagnostic screening by sequencing. However, sequencing is a time-consuming and complicated procedure, not suitable for routine clinical use.Experimental Design: We have developed rapid, simple, and sensitive mutation detection assays based on the SMart Amplification Process (SMAP) and applied it for analyzing EGFR gene mutations in clinical samples. By using SMAP, we can detect mutations within 30 min including sample preparation. To validate the assay system for potential use in clinical diagnostics, we examined 45 NSCLC patients for EGFR mutations using sequencing and SMAP.Results: The outcomes of the SMAP assay perfectly matched the Sequencing results, except in one case where SMAP was able to identify a mutation that was not detected by sequencing. We also evaluated the sensitivity and specificity of SMAP in mutation detection for EGFR. In a serial dilution study, SMAP was able to find a mutation in a sample containing only 0.1% of the mutant allele in a mixture of wild-type genomic DNA. We also could show amplification of mutated DNA with only 30 copies per reaction.Conclusions: The SMAP method offers higher sensitivity and specificity than alternative technologies, while eliminating the need for sequencing to identify mutations in the EGFR gene of NSCLC. It provides a robust and point-of-care accessible approach for a rapid identification of most patients likely to respond to gefitinib.