Journal of Clinical Virology 44 (2009) 157–160 Contents lists available at ScienceDirect Journal of Clinical Virology journal homepage: www.elsevier.com/locate/jcv Short communication Genotyping of acute HBV isolates from England, 1997–2001 Richard D. Sloan a,∗ , Angela L. Strang a , Mary E. Ramsay b , Chong-Gee Teo a a Virus Reference Division, Centre for Infections, Health Protection Agency, Colindale Avenue, London NW9 5HT, UK b Communicable Disease Surveillance Centre, Centre for Infections, Health Protection Agency, Colindale Avenue, London NW9 5HT, UK article info Article history: Received 23 April 2008 Received in revised form 19 November 2008 Accepted 24 November 2008 Keywords: Hepatitis B virus Acute viral infection Viral genotype Phylogeny abstract Background: Increasing data shows the relevance of HBV genotypes in the outcome of infection. Most studies investigating the relationship between the genotypic characteristics of hepatitis B virus (HBV) and the clinical or epidemiological aspects of HBV infection originate from studies of patients with chronic rather than acute hepatitis B. Objectives: To study a convenience sample representing ca. 5% of reported acute hepatitis B in England between 1997 and 2001 to investigate the distribution of HBV genotypes and specific HBV variants with epidemiological risk factors, thereby providing baseline data for ongoing surveillance. Study design: From 160 serum samples, PCR was carried out to amplify the first 600 bases of the HBV S gene. Amplicons were sequenced and subjected to phylogenetic analysis and risk factor analysis. Results: Fifty-seven percent of the study samples carried HBV belonging to subtype A2, 13% to subtype D2, and the rest to genotype E (8%) and subtypes C2 and D3 (each 6%), D1 and D4 (each 3%) and B4 (1%). One particular A2 isolate was dominant, accounting for 23% of the total sample set. Drug use and homosexual transmission were equally implicated as risks within genotype A2. No mutations associated with vaccine escape or resistance to antiviral therapy were identified. Conclusion: Immigration and travel likely shape the observed genotype distribution and consequent prevalence of genotypes other than A2 or D in this population. Data suggests no genetic separation of parenteral and sexually transmitted virus. These data demonstrate the value in pursuing more extensive and recent surveillance. © 2008 Elsevier B.V. All rights reserved. 1. Introduction Hepatitis B virus (HBV) has been classified into 8 genotypes, designated A to G. These genotypes have been associated with outcomes of chronic hepatitis B such as progression to severe disease and development of liver cancer, and response to antivi- ral therapy based on interferons and analogues of nucleotides or nucleosides. 1,2 Knowledge of the HBV genotype distribution in patients with chronic hepatitis B may aid the formulation and implementation of initiatives to lessen the morbidity and mortal- ity burden with which it is associated. Studies in Europe enquiring into the distribution of HBV genotypes have been helpful in this regard. 3–5 From a broader public health perspective, it would be important also to determine, in acutely infected people, how par- ticular genotypes and strains of HBV are being transmitted within specific risk groups. Presented here are findings from an inves- tigation to describe HBV genotypes, epidemiological risk factors ∗ Corresponding author at: McGill AIDS Centre, Lady Davis Institute, 3755 Côte- Ste-Catherine, Montréal, Québec H3T 1E2, Canada. Tel.: +1 514 340 8222; fax: +1 514 340 7537. E-mail address: richard.sloan@mail.mcgill.ca (R.D. Sloan). and specific HBV variants associated with acute hepatitis B in Eng- land. 2. Materials and methods Study samples were assembled from hepatitis B surface antigen-positive sera of patients with suspected acute hepatitis B testing between January 1997 and December 2001 to the Virus Reference Division (VRD) of the Centre for Infections of the Health Protection Agency (HPA). All samples had been referred to the HPA for tertiary reference analysis. Samples were included into the study if they had been tested by the VRD to contain >200 Paul Erlich Institute International Units per ml of IgM to the anti-hepatitis B core antigen. Two hundred and twenty four samples satisfied this criterion and were thus included for analysis. Risk factors (where available), patient gender and age were also recorded from patient records provided by referring clinicians and by screening of HPA epidemiological databases. From these samples, DNA from the HBV S gene was amplified using nested PCR. Primers for first-round PCR were 5 ′ -AGCCCTCAGGCTCAGGGCATA-3 ′ (outer, sense) and 5 ′ -AAACCCAGAAGACCCACA-3 ′ (outer, antisense), and the second-round primers were 5 ′ -TCATCCTCAGGCCATGCAGT-3 ′ (inner, sense) and 5 ′ -ACACACTTTCCAATCAATAG-3 ′ (outer, sense). 1386-6532/$ – see front matter © 2008 Elsevier B.V. All rights reserved. doi:10.1016/j.jcv.2008.11.010