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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter / Tate, Jillian R.

12 Issues per year

IMPACT FACTOR 2016: 3.432

CiteScore 2016: 2.21

SCImago Journal Rank (SJR) 2016: 1.000
Source Normalized Impact per Paper (SNIP) 2016: 1.112

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Volume 48, Issue 8 (Aug 2010)


Development and evaluation of an automated hepatitis C virus NS5B sequence-based subtyping assay

Diana Koletzki / Stéphanie Dumont / Hans Vermeiren / Bart Fevery / Pieter De Smet / Lieven J. Stuyver
Published Online: 2010-06-28 | DOI: https://doi.org/10.1515/CCLM.2010.236


Background: Hepatitis C virus (HCV) genotyping and accurate subtype determination is becoming increasingly important to better understand viral evolution, the development of resistance to STAT-C, and possibly even for the treatment and management of chronic HCV-infected patients.

Methods: A subtyping assay based on a 329-bp sequence of the NS5B region, together with an automated subtype interpretation tool was developed. Clinical samples of the six major genotypes were used to assess assay performance characteristics.

Results: The NS5B BLAST-based subtyping assay showed clinical sensitivity for amplification of 89% (n=603 random samples). Assessment of analytical sensitivity of amplification for genotypes 1, 2, 3 and 4 revealed a suitable performance for high viral load samples and decreased only with low viral loads. The results were 100% and 99% accurate at the genotype and subtype level, respectively. A concordance of 97% on genotype level and 62% on subtype level was observed by comparison with subtype results from 5′ non-coding-based assays with a panel of 276 isolates.

Conclusions: The HCV NS5B subtyping assay has been validated for research use. Based on its performance, it is the method of choice in cases where subtype rather than genotype information is needed.

Clin Chem Lab Med 2010;48:1095–102.

Keywords: hepatitis C virus; NS5B; sequencing; subtype

About the article

Corresponding author: Dr. Diana Koletzki, Virco BVBA, Generaal De Wittelaan L11 B3, 2800 Mechelen, Belgium Phone: +32 15 461 356, Fax: +32 15 461 955,

Received: 2009-08-14

Accepted: 2010-03-11

Published Online: 2010-06-28

Published in Print: 2010-08-01

Citation Information: Clinical Chemistry and Laboratory Medicine, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/CCLM.2010.236.

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©2010 by Walter de Gruyter Berlin New York. Copyright Clearance Center

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