Prospective study of the accuracy of EGFR mutational analysis by high-resolution melting analysis in small samples obtained from patients with non-small cell lung cancer
Prospective study of the accuracy of EGFR mutational analysis by high-resolution melting analysis in small samples obtained from patients with non-small cell lung cancer
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DOI：
10.1158/1078-0432.ccr-07-5207
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发表时间：
2008-08-01
影响因子：
11.5
通讯作者：
Tamura, Tomohide
中科院分区：
文献类型：
--
作者：
Fukui, Tomoya;Ohe, Yuichiro;Tamura, Tomohide
目的：表皮生长因子受体（EGFR）突变，特别是外显子19（DEL）的框内缺失和外显子21（L858R）的点突变，可预测非小细胞肺癌（NSCLC）患者对吉非替尼的敏感性。在这项研究中，我们验证了高分辨率熔解分析（HRMA），这是一种快速的方法，使用PCR扩增与染料分析的熔解曲线在NSCLC的小样本EGFR突变分析的准确性。实验设计：我们设计了一项前瞻性研究，比较HRMA和DNA测序与激光捕获显微切割的灵敏度和特异性。通过支气管镜检查或经皮穿刺活检筛选符合条件的肺部病变患者，以组织学确诊，然后手术切除NSCLC。采用HRMA检测小样本EGFR突变，采用HRMA和DNA测序检测手术切除标本EGFR突变。18例（34.6%）患者检测到EGFR突变。手术切除标本的HRMA和DNA测序结果显示100%的敏感性和特异性。另一方面，HRMA从小诊断标本的敏感性和特异性分别为83.3%和100%，respectives.Conclusions：在这项研究中，我们表明，HRMA是一个高度准确的方法检测DEL和L858R突变的NSCLC患者，虽然有必要考虑识别的患者与假阴性结果时，进行分析，使用小样本。
Purpose: Epidermal growth factor receptor (EGFR) mutations, especially in-frame deletions in exon 19 (DEL) and a point mutation in exon 21 (L858R), predict gefitinib sensitivity in patients with non-small cell lung cancer (NSCLC). In this study, we verified the accuracy of EGFR mutation analysis in small samples by high-resolution melting analysis (HRMA), which is a rapid method using PCR amplification with a dye to analyze the melting curves in NSCLC.Experimental Design: We designed a prospective study to compare the sensitivity and specificity of HRMA and DNA sequencing with laser capture microdissection. Eligible patients with lung lesions were screened by bronchoscopy or percutaneous needle biopsy to histologically confirm the diagnosis, followed by surgical resection of the NSCLC. Small diagnostic specimens were analyzed for EGFR mutations by HRMA, and the surgically resected specimens were examined for mutations by HRMA and DNA sequencing.Results: The analyses for EGFR mutations were conducted in 52 eligible cases of the 92 enrolled patients. EGFR mutations were detected in 18 (34.6%) patients. The results of HRMA from surgically resected specimens as well as DNA sequencing revealed 100% sensitivity and specificity. On the other hand, the sensitivity and specificity of HRMA from the small diagnostic specimens were 83.3 % and 100%, respectively.Conclusions: In this study, we showed that HRMA is a highly accurate method for detecting DEL and L858R mutations in patients with NSCLC, although it is necessary to consider the identification of patients with a false-negative result when the analysis is conducted using small samples.