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Journal of Clinical Microbiology, August 2006, p. 2779-2784, Vol. 44, No. 8
0095-1137/06/$08.00+0     doi:10.1128/JCM.01638-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Multitypic Hepatitis C Virus Infection Identified by Real-Time Nucleotide Sequencing of Minority Genotypes

Andrew J. Buckton,1* Siew-Lin Ngui,1 Catherine Arnold,1 Katie Boast,2 Joanne Kovacs,2 Paul E. Klapper,3 Bharat Patel,4 Imad Ibrahim,5 Savita Rangarajan,2 Mary E. Ramsay,1 and Chong-Gee Teo1

Centre for Infections, Health Protection Agency, Colindale Avenue, London, United Kingdom,1 Haemophilia Reference Centre, St. Thomas' Hospital, London, United Kingdom,2 West Yorkshire Health Protection Unit, Bridle Path, York Road, Leeds, United Kingdom,3 HPA Collaborating Laboratory, Department of Microbiology, North Middlesex University Hospital, London, United Kingdom,4 Microbiology Department, Ealing Hospital, Uxbridge Road, Southall, Middlesex, United Kingdom5

Received 9 August 2005/ Returned for modification 4 November 2005/ Accepted 12 June 2006

The prevalence of concurrent multitypic hepatitis C virus (HCV) infection is uncertain. A sensitive and specific approach to identifying minority HCV genotypes in blood is presented. Following serum extraction and reverse transcription PCR to amplify cDNA originating from the viral 5' noncoding region, the amplified product mixture was treated with genotype-specific restriction endonuclease to digest the dominant genotype. Residual amplicons were subjected to PCR cloning and then to real-time DNA sequencing using a Pyrosequencer to identify the remaining genotypes. Dilution experiments showed that minority genotypes may be detected when they represent 1:10,000 of the total population and in serum specimens with viral loads as low as 1,000 IU/ml. Of 37 patients with bleeding disorders and 44 injecting drug users, infection by more than one HCV genotype was found in 7 (19%) and 4 (9%) patients, respectively. The low rate of detection in people at high risk of repeated HCV infection suggests that multitypic HCV carriage is uncommon.

* Corresponding author. Mailing address: Centre for Infections, Health Protection Agency, 61 Colindale Avenue, Colindale, London NW9 5HT, United Kingdom. E-mail: andrew.buckton{at}hpa.org.uk.

Journal of Clinical Microbiology, August 2006, p. 2779-2784, Vol. 44, No. 8
0095-1137/06/$08.00+0     doi:10.1128/JCM.01638-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Martro, E., Gonzalez, V., Buckton, A. J., Saludes, V., Fernandez, G., Matas, L., Planas, R., Ausina, V. (2008). Evaluation of a New Assay in Comparison with Reverse Hybridization and Sequencing Methods for Hepatitis C Virus Genotyping Targeting Both 5' Noncoding and Nonstructural 5b Genomic Regions. J. Clin. Microbiol. 46: 192-197 [Abstract] [Full Text]  


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