Suppr超能文献

基于聚合酶链反应(PCR)检测非小细胞肺癌患者中与治疗相关的表皮生长因子受体(EGFR)突变

PCR-based testing for therapy-related EGFR mutations in patients with non-small cell lung cancer.

作者信息

Dahse Regine, Berndt Alexander, Kosmehl Hartwig

机构信息

HELIOS Clinic Erfurt, Institute of Pathology, Erfurt, Germany.

出版信息

Anticancer Res. 2008 Jul-Aug;28(4B):2265-70.

Abstract

BACKGROUND

In patients with non-small cell lung cancer (NSCLC), mutations in the epidermal growth factor receptor gene (EGFR) have been associated with improved response to tyrosine kinase inhibitors. Two hotspot mutations located in exon 19 and exon 21 account for about 90% of all EGFR mutations.

MATERIALS AND METHODS

We designed a Bi-PASA (bidirectional PCR amplification of specific alleles) assay to detect the most common exon 19 deletion (codons 746-750) and an allele-specific PCR for the L858R mutation in exon 21. To validate the assays for use in clinical diagnostics, 35 lung adenocarcinoma samples were analyzed.

RESULTS

Both assays provided the predicted amplification pattern for normal and mutant genotypes with high specificity and sensitivity. In serial dilution experiments, the mutant alleles were detectable in mixed samples with an at least 6-fold excess of normal DNA. Three exon 19 deletions were identified in the tumor samples.

CONCLUSION

Both assays are fast and easy to perform in any routine PCR laboratory with no special equipment other than thermocyclers. They provide sensitive and cost effective initial EGFR testing for identifying lung cancer patients who might clinically benefit from tyrosine kinase inhibitors.

摘要

背景

在非小细胞肺癌(NSCLC)患者中,表皮生长因子受体基因(EGFR)突变与对酪氨酸激酶抑制剂的反应改善相关。位于外显子19和外显子21的两个热点突变约占所有EGFR突变的90%。

材料与方法

我们设计了一种双向等位基因特异性PCR(Bi-PASA)检测方法,用于检测最常见的外显子19缺失(密码子746-750),并设计了一种针对外显子21中L858R突变的等位基因特异性PCR。为验证这些检测方法在临床诊断中的应用,我们分析了35份肺腺癌样本。

结果

两种检测方法均为正常和突变基因型提供了预测的扩增模式,具有高特异性和高灵敏度。在系列稀释实验中,在正常DNA至少过量6倍的混合样本中可检测到突变等位基因。在肿瘤样本中鉴定出3例外显子19缺失。

结论

两种检测方法在任何常规PCR实验室都能快速、简便地进行,除热循环仪外无需特殊设备。它们为识别可能从酪氨酸激酶抑制剂中临床获益的肺癌患者提供了灵敏且经济高效的初始EGFR检测。

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验