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Genes 2015, 6(1), 46-59; doi:10.3390/genes6010046

Applicability of Next Generation Sequencing Technology in Microsatellite Instability Testing

1
Department of Pathology, Faculty of Medicine, Dentistry and Health Sciences, University of Melbourne, Parkville, Victoria 3052, Australia
2
Department of Colorectal Medicine and Genetics, Familial Cancer Clinic, Royal Melbourne Hospital, Parkville, Victoria 3050, Australia
3
Department of Medicine, Royal Melbourne Hospital, University of Melbourne, Parkville, Victoria 3050, Australia
4
Department of Pathology and Sir Peter MacCallum Department of Oncology, University of Melbourne, East Melbourne, Victoria 3002, Australia
*
Author to whom correspondence should be addressed.
Academic Editor: Maija Kohonen-Corish
Received: 25 November 2014 / Accepted: 27 January 2015 / Published: 12 February 2015
(This article belongs to the Special Issue Microsatellite Instability)
View Full-Text   |   Download PDF [7772 KB, uploaded 12 February 2015]   |  

Abstract

Microsatellite instability (MSI) is a useful marker for risk assessment, prediction of chemotherapy responsiveness and prognosis in patients with colorectal cancer. Here, we describe a next generation sequencing approach for MSI testing using the MiSeq platform. Different from other MSI capturing strategies that are based on targeted gene capture, we utilize “deep resequencing”, where we focus the sequencing on only the microsatellite regions of interest. We sequenced a series of 44 colorectal tumours with normal controls for five MSI loci (BAT25, BAT26, BAT34c4, D18S55, D5S346) and a second series of six colorectal tumours (no control) with two mononucleotide loci (BAT25, BAT26). In the first series, we were able to determine 17 MSI-High, 1 MSI-Low and 26 microsatellite stable (MSS) tumours. In the second series, there were three MSI-High and three MSS tumours. Although there was some variation within individual markers, this NGS method produced the same overall MSI status for each tumour, as obtained with the traditional multiplex PCR-based method. View Full-Text
Keywords: microsatellite instability; next generation sequencing; colorectal cancer; MiSeq microsatellite instability; next generation sequencing; colorectal cancer; MiSeq
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Gan, C.; Love, C.; Beshay, V.; Macrae, F.; Fox, S.; Waring, P.; Taylor, G. Applicability of Next Generation Sequencing Technology in Microsatellite Instability Testing. Genes 2015, 6, 46-59.

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