Performance of a RT-PCR Assay in Comparison to FISH and Immunohistochemistry for the Detection of ALK in Non-Small Cell Lung Cancer

dc.contributor.authorHout, David R.
dc.contributor.authorSchweitzer, Brock L.
dc.contributor.authorLawrence, Kasey
dc.contributor.authorMorris, Stephan W.
dc.contributor.authorTucker, Tracy
dc.contributor.authorMazzola, Rosetta
dc.contributor.authorSkelton, Rachel
dc.contributor.authorMcMahon, Frank
dc.contributor.authorHandshoe, John
dc.contributor.authorLesperance, Mary
dc.contributor.authorKarsan, Aly
dc.contributor.authorSaltman, David L.
dc.description.abstractPatients with lung cancers harboring an activating anaplastic lymphoma kinase (ALK) rearrangement respond favorably to ALK inhibitor therapy. Fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC) are validated and widely used screening tests for ALK 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 ALK 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 ΔCt of ≤8) was shown to be highly sensitive (100%) when compared to FISH and IHC. Sequencing of RNA detected full-length ALK transcripts or EML4-ALK and KIF5B-ALK fusion variants in discordant cases in which ALK 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 ALK 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.en_US
dc.description.sponsorshipThis work was supported through a National Cancer Institute of the United States contract: SBIR Topic 277 Phase II, entitled “Development of a Companion Diagnostic for ALK Mutations”. Contract No.: HHSN261201200047C. Administrative assistance for the conduct of the study was provided by Helena Daudt and Jodi Leblanc. The authors thank Diana Ionescu, Tadaaki Hiruki, and Chen Zhou of the Department of Pathology and Laboratory Medicine, BC Cancer Agency, and the pathologists from referring hospitals who participated in this study.en_US
dc.identifier.citationHout, D.R., Schweitzer, B.L., Lawrence, K., Morris, S.W., Tucker, T., Mazzola, R.,… Saltman, D.L. (2017). Performance of a RT-PCR Assay in Comparison to FISH and Immunohistochemistry for the Detection of ALK in Non-Small Cell Lung Cancer. Cancers, 9(8), 99.
dc.subjectanaplastic lymphoma kinaseen_US
dc.subjectfluorescence in situ hybridizationen_US
dc.subjectnon-small cell lung canceren_US
dc.subjectreverse transcriptase-polymerase chain reactionen_US
dc.titlePerformance of a RT-PCR Assay in Comparison to FISH and Immunohistochemistry for the Detection of ALK in Non-Small Cell Lung Canceren_US


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