Hout David R, Schweitzer Brock L, Lawrence Kasey, Morris Stephan W, Tucker Tracy, Mazzola Rosetta, Skelton Rachel, McMahon Frank, Handshoe John, Lesperance Mary, Karsan Aly, Saltman David L
Insight Genetics, Inc., Suite 510, 2 International Plaza, Nashville, TN 37217, USA.
Department of Pathology and Laboratory Medicine, BC Cancer Agency, 675 West 10th Avenue, Vancouver, BC V5Z 1L3, Canada.
Cancers (Basel). 2017 Aug 1;9(8):99. doi: 10.3390/cancers9080099.
Patients with lung cancers harboring an activating anaplastic lymphoma kinase () rearrangement respond favorably to ALK inhibitor therapy. Fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC) are validated and widely used screening tests for rearrangements but both methods have limitations. The ALK RGQ RT-PCR Kit (RT-PCR) is a single tube quantitative real-time PCR assay for high throughput and automated interpretation of expression. In this study, we performed a direct comparison of formalin-fixed paraffin-embedded (FFPE) lung cancer specimens using all three ALK detection methods. The RT-PCR test (diagnostic cut-off Δ of ≤8) was shown to be highly sensitive (100%) when compared to FISH and IHC. Sequencing of RNA detected full-length transcripts or and fusion variants in discordant cases in which expression was detected by the ALK RT-PCR test but negative by FISH and IHC. The overall specificity of the RT-PCR test for the detection of ALK in cases without full-length expression was 94% in comparison to FISH and sequencing. These data support the ALK RT-PCR test as a highly efficient and reliable diagnostic screening approach to identify patients with non-small cell lung cancer whose tumors are driven by oncogenic ALK.
携带激活型间变性淋巴瘤激酶（ALK）重排的肺癌患者对ALK抑制剂治疗反应良好。荧光原位杂交（FISH）和免疫组织化学（IHC）是经过验证且广泛应用于ALK重排的筛查检测方法，但这两种方法都有局限性。ALK RGQ逆转录聚合酶链反应试剂盒（RT-PCR）是一种单管定量实时PCR检测方法，用于高通量自动解读ALK表达。在本研究中，我们使用所有三种ALK检测方法对福尔马林固定石蜡包埋（FFPE）肺癌标本进行了直接比较。与FISH和IHC相比，RT-PCR检测（诊断临界值Δ≤8）显示出高度敏感性（100%）。在ALK RT-PCR检测显示ALK表达但FISH和IHC检测为阴性的不一致病例中，RNA测序检测到全长ALK转录本或ALK和EML4融合变体。与FISH和测序相比，RT-PCR检测在无全长ALK表达病例中检测ALK的总体特异性为94%。这些数据支持ALK RT-PCR检测作为一种高效可靠的诊断筛查方法，用于识别肿瘤由致癌性ALK驱动的非小细胞肺癌患者。
