Detection of EML4-ALK in serum RNA from lung cancer patients

在非小细胞肺癌(NSCLC)中，转化融合基因EML4-ALK的存在是预测ALK激酶抑制剂疗效的一个指标。血清循环RNA是无创性分子诊断的一个新兴领域。我们利用基于PCR的MassARRAY平台设计了EML4-ALK检测方法。我们尝试了一种RNA提取方法、引物设计和逆转录试剂来优化我们的血清样本检测系统。我们评估了12例ALK重排的非小细胞肺癌患者血清中是否能检测到EML4-ALK融合RNA。12例患者中有2例检测到EML4-ALK RNA(变异体1和变异体3)。用Massarray法检测血清RNA样品中的EML4-ALK是可行的。对于肿瘤标本不足或无法获得的患者，这种非侵入性检测将是有用的。更多的实验将揭示该检测系统的敏感性和特异性。

The presence of the transforming fusion gene EML4-ALK in non–small cell lung cancer (NSCLC) is a predictive marker for the efficacy of ALK kinase inhibitors. Circulating RNA in serum is an emerging field for noninvasive molecular diagnosis. We designed EML4-ALK detection assay using PCR-based MassARRAY platform. We attempted a RNA extraction method, primer design, and reverse transcription reagents to optimize our detection system with serum samples. We evaluated whether EML4-ALK fusion RNA could be detected in serum obtained from 12 NSCLC patients with ALK rearrangements. The EML4-ALK RNA (variant 1 and 3) was detected in serum from 2 of 12 patients. The detection of EML4-ALK in serum RNA samples by MassARRAY assay is feasible. This noninvasive assay will be useful in patients with insufficient or unavailable tumor specimens. Additional experiments will reveal the sensitivity and specificity of this detection system.