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Journal of Clinical Microbiology, January 2004, p. 158-163, Vol. 42, No. 1
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.1.158-163.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Hepatitis C Genotyping by Denaturing High-Performance Liquid Chromatography

Michael Liew,1* Maria Erali,1 Sam Page,1 David Hillyard,1,2 and Carl Wittwer1,2

Institute for Clinical and Experimental Pathology, ARUP Laboratories, Salt Lake City, Utah 84108-1221,1 Department of Pathology, School of Medicine, University of Utah, Salt Lake City, Utah 841322

Received 7 May 2003/ Returned for modification 21 June 2003/ Accepted 12 October 2003

Determination of the hepatitis C virus (HCV) genotype for infected patients increasingly has become accepted as the standard of care. Genotype assignment helps in assessing disease prognosis and assists in establishing the appropriate duration of treatment. The great genetic diversity of HCV, with 11 major genotypes and >70 subtypes, contributes to the technical difficulty of genotype testing. While the "gold standard" for testing is nucleic acid sequencing, a variety of hybridization assays, including the line probe assay, have been developed to provide more rapid and accessible forms of testing. The aim of this study was to determine whether denaturing high-performance liquid chromatography (dHPLC) could be used as a clinical method for distinguishing HCV genotypes 1, 2, 3, and 4. A portion of the 5' untranslated region of the HCV genome was amplified by heminested multiplex reverse transcription PCR. The two amplicons then were analyzed by dHPLC analysis and compared to the genotypes determined by sequence analysis. After 115 specimens were analyzed as standards, 200 masked specimens (specimens whose identity was not known before testing) were analyzed to determine the concordance of the assay. The assay had a concordance of 96% at the genotype level and a concordance of 87% at the subtype level. However, the dHPLC method was not as accurate as other reported methods of HCV genotyping. This is the first time that HCV genotyping has been performed by dHPLC.

* Corresponding author. Mailing address: ARUP Laboratories, Institute for Clinical and Experimental Pathology, 500 Chipeta Way, Salt Lake City, UT 84108-1221. Phone: (801) 583-2787, ext. 2179. Fax: (801) 584-5114. E-mail: liewm{at}aruplab.com.

Journal of Clinical Microbiology, January 2004, p. 158-163, Vol. 42, No. 1
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.1.158-163.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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