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Journal of Clinical Microbiology, August 2008, p. 2723-2730, Vol. 46, No. 8
0095-1137/08/$08.00+0     doi:10.1128/JCM.01622-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Application of a Novel, Rapid, and Sensitive Oligonucleotide Ligation Assay for Detection of Cancer-Predicting Mutations in the Precore and Basal Core Promoter of Hepatitis B Virus{triangledown}

M. E. Mendy,1* S. Kaye,1,{dagger} E. Le Roux,2 G. D. Kirk,3 A. Jeng-Barry,1 S. McConkey,1,{ddagger} M. Cotten,1 M. H. Kuniholm,3 A. Leligdowicz,1 P. Hainaut,2 S. Rowland-Jones,1 and H. Whittle1

Viral Diseases Programme, Medical Research Council, Banjul, The Gambia,1 Molecular Carcinogenesis, International Agency for Research on Cancer, Lyon, France,2 Department of Epidemiology, Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland3

Received 14 August 2007/ Returned for modification 21 November 2007/ Accepted 17 May 2008

Hepatocellular carcinoma (HCC) and cirrhosis are important causes of mortality worldwide. Persistent hepatitis B virus (HBV) infection is a major cause of these diseases. Double mutations in the basal core promoter (BCP) (A1762T and G1764A) and precore (pre-C) (G1896A) regions of the virus are associated with progression to HCC. The current study is aimed at developing a simple method for screening and detecting BCP and pre-C mutations in HBV carriers. We have developed and validated an oligonucleotide ligation assay (OLA) to detect point mutations in the HBV core gene. We have applied OLA methods to samples from HBV-infected carriers recruited from the Gambia Liver Cancer Study (GLCS) comprising asymptomatic HBsAg carriers, patients with cirrhosis, and patients with HCC. We observed an 89.3% and 95.8% concordance between the OLA and DNA sequencing for BCP and pre-C mutations, respectively. OLA detected the mutations in single-strain infections and in infections with mixtures of wild-type and mutant viruses under conditions where sequencing detected only the single dominant strains. BCP mutations were detected in 75.7% of patients with advanced liver disease (cirrhosis/HCC) compared to 47.6% of asymptomatic carriers, while pre-C mutations were detected in 34.5% of advanced liver disease patients and in 47.6% of asymptomatic HBsAg carriers. There was a significant association between the presence of BCP mutations and advanced liver disease. In conclusion, OLA is a simple, economical, and reliable assay for detection of pre-C and BCP mutations. Its application can lead to improvement in diagnosis and clinical care in regions where HBV is endemic.

* Corresponding author. Mailing address: Viral Diseases Programme, Medical Research Council, Atlantic Boulevard, Fajara, P.O. Box 273, Banjul, The Gambia. Phone: 220 4495442. Fax: 220 4495919. E-mail: mmendy{at}mrc.gm

{triangledown} Published ahead of print on 28 May 2008.

{dagger} Present address: Imperial College, London, United Kingdom.

{ddagger} Present address: Royal College of Surgeons in Ireland, Dublin, Ireland.

Journal of Clinical Microbiology, August 2008, p. 2723-2730, Vol. 46, No. 8
0095-1137/08/$08.00+0     doi:10.1128/JCM.01622-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Liu, S., Zhang, H., Gu, C., Yin, J., He, Y., Xie, J., Cao, G. (2009). Associations Between Hepatitis B Virus Mutations and the Risk of Hepatocellular Carcinoma: A Meta-Analysis. JNCI J Natl Cancer Inst 101: 1066-1082 [Abstract] [Full Text]  


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