Kikuchi, Hugh, Reiman, Anne, Nyoni, Jenifer, Lloyd, Katherine L., Savage, Richard S., Wotherspoon, Tina, Berry, Lisa, Snead, David and Cree, Ian A. (2016) Development and validation of a TaqMan Array for cancer mutation analysis. Pathogenesis, 3 (1). pp. 1-8. S2214-6636(16)00002-X. doi:10.1016/j.pathog.2016.02.001 ISSN 2214-6636.

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Abstract

Introduction:

Optimal cancer treatment with targeted agents requires rapid, comprehensive and accurate molecular assays to analyse actionable oncogenic mutations across multiple tumour types.

Materials and Methods:

We describe a PCR panel based on the 384 well TaqMan Array® (Thermo Fisher Scientific). This allows measurement of common RAS (NRAS and KRAS), EGFR and BRAF mutations in a single assay (the REB Array), analysing 44 mutations in 7 samples per plate. This retrospective study includes 96 patients with NSCLC (n = 42), colorectal cancer (n = 26), and melanoma (n = 28) with previous mutational analysis. Samples with discrepant results were sequenced to confirm the result.

Results:

The REB achieved 93% concordance with the Therascreen EGFR assay (Qiagen), 95% concordance with the KRAS castPCR assay (Thermo Fisher), and 100% concordance with the cobas BRAF assay (Roche). There were 2 true discrepancies, most likely a result of sample quality or differences in sensitivity between the assays that depend on set thresholds to determine the presence of mutations. Analysis of the performance of the REB Array gave an overall sensitivity of 92%, with a positive predictive value of 100% and negative predictive value of 84.24%.

Conclusion:

The REB array is comparable to competing PCR methods with the additional advantages of a broader range of mutations, simplified manual handling, and reduced overall cost per sample.

Item Type:	Journal Article
Subjects:	R Medicine > RC Internal medicine > RC0254 Neoplasms. Tumors. Oncology (including Cancer)
Divisions:	Faculty of Science, Engineering and Medicine > Medicine > Warwick Medical School > Biomedical Sciences > Translational & Experimental Medicine > Reproductive Health ( - until July 2016) Faculty of Science, Engineering and Medicine > Science > Statistics Faculty of Science, Engineering and Medicine > Research Centres > Warwick Systems Biology Centre Faculty of Science, Engineering and Medicine > Medicine > Warwick Medical School > Biomedical Sciences > Translational & Experimental Medicine Faculty of Science, Engineering and Medicine > Medicine > Warwick Medical School
Library of Congress Subject Headings (LCSH):	Cancer -- Treatment
Journal or Publication Title:	Pathogenesis
Publisher:	Elsevier BV
ISSN:	2214-6636
Official Date:	February 2016
Dates:	Date Event February 2016 Published 26 February 2016 Available 19 February 2016 Accepted 29 January 2016 Submitted
Volume:	3
Number:	1
Number of Pages:	8
Page Range:	pp. 1-8
Article Number:	S2214-6636(16)00002-X
DOI:	10.1016/j.pathog.2016.02.001
Status:	Peer Reviewed
Publication Status:	Published
Access rights to Published version:	Open Access (Creative Commons)
Funder:	CanTech Ltd., National Institute for Health Research (Great Britain) (NIHR), University of Warwick

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