Multitypic hepatitis C virus infection identified by real-time nucleotide sequencing of minority genotypes

Andrew J. Buckton*, Siew Lin Ngui, Catherine Arnold, Katie Boast, Joanne Kovacs, Paul E. Klapper, Bharat Patel, Imad Ibrahim, Savita Rangarajan, Mary Ramsay, Chong Gee Teo

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

21 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)2779-2784
Number of pages6
JournalJournal of Clinical Microbiology
Volume44
Issue number8
DOIs
Publication statusPublished - Aug 2006

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