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Cancers 2017, 9(8), 99; doi:10.3390/cancers9080099

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

1
Insight Genetics, Inc., Suite 510, 2 International Plaza, Nashville, TN 37217, USA
2
Department of Pathology and Laboratory Medicine, BC Cancer Agency, 675 West 10th Avenue, Vancouver, BC V5Z 1L3, Canada
3
Department of Medical Oncology, British Columbia Cancer Agency, VIC 2410 Lee Avenue, Victoria, BC V8R 6V5, Canada
4
Department of Mathematics and Statistics, University of Victoria, Box 1700, STN CSC, Victoria, BC V8W 2Y2, Canada
*
Author to whom correspondence should be addressed.
Academic Editor: Raymond Lai
Received: 15 June 2017 / Revised: 19 July 2017 / Accepted: 29 July 2017 / Published: 1 August 2017
(This article belongs to the Special Issue Targeting ALK in Cancer)
View Full-Text   |   Download PDF [681 KB, uploaded 1 August 2017]   |  

Abstract

Patients 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. View Full-Text
Keywords: anaplastic lymphoma kinase; fluorescence in situ hybridization; immunohistochemistry; non-small cell lung cancer; reverse transcriptase-polymerase chain reaction anaplastic lymphoma kinase; fluorescence in situ hybridization; immunohistochemistry; non-small cell lung cancer; reverse transcriptase-polymerase chain reaction
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MDPI and ACS Style

Hout, D.R.; Schweitzer, B.L.; Lawrence, K.; Morris, S.W.; Tucker, T.; Mazzola, R.; Skelton, R.; McMahon, F.; Handshoe, J.; Lesperance, M.; Karsan, A.; Saltman, D.L. Performance of a RT-PCR Assay in Comparison to FISH and Immunohistochemistry for the Detection of ALK in Non-Small Cell Lung Cancer. Cancers 2017, 9, 99.

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