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Journal of Clinical Microbiology, June 2002, p. 2046-2050, Vol. 40, No. 6
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.6.2046-2050.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Genotyping of Hepatitis C Virus Types 1, 2, 3, and 4 by a One-Step LightCycler Method Using Three Different Pairs of Hybridization Probes

Matthias Schröter,1 Bernhard Zöllner,1 Peter Schäfer,1 Olfert Landt,2 Ulrich Lass,2 Rainer Laufs,1 and Heinz-Hubert Feucht1*

Institut für Medizinische Mikrobiologie und Immunologie, Universitätsklinikum Hamburg-Eppendorf, 20246 Hamburg,1 TIB MOLBIOL, 10829 Berlin, Germany2

Received 10 July 2001/ Returned for modification 24 November 2001/ Accepted 20 January 2002

Determination of hepatitis C virus (HCV) genotypes has become increasingly important during the last years for prediction of the clinical course and the outcome of antiviral therapy. Therefore, numerous different methods have been developed to enable HCV genotyping. However, many of them are very laborious and expensive, leading to limited usage in daily routine diagnostics. We have established a method which combines the speed of the new LightCycler technology with the use of amplification products generated for diagnostic quantitative HCV RNA determination. Differentiation of HCV genotypes is performed with these amplicons in a single step by using fluorophore-labeled hybridization probes. Although currently only two different acceptor fluorophores are available for the LightCycler, types 1, 2, 3, and 4, which are by far the prevailing HCV genotypes in Europe and the United States, can be distinguished. Genotypes of specimens from 190 chronically HCV-infected patients were determined by the LightCycler method and compared with the results of nucleotide sequencing. Concordant results were obtained for all samples. This new method offers a fast and convenient possibility to determine the quantitative HCV RNA load and the genotype in large-scale settings within about 4 h.

* Corresponding author: Mailing address: Institut für Medizinische Mikrobiologie und Immunologie, Universitätsklinikum Hamburg-Eppendorf, Martinistrasse 52, 20246 Hamburg, Germany. Phone: 49 (0)40 42803 3159. Fax: 49 (0)40 42803 4062. E-mail: feucht{at}uke.uni-hamburg.de.

Journal of Clinical Microbiology, June 2002, p. 2046-2050, Vol. 40, No. 6
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.6.2046-2050.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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