This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Osiowy, C. Search for Related Content PubMed PubMed Citation Articles by Osiowy, C.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, July 2002, p. 2566-2571, Vol. 40, No. 7
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.7.2566-2571.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Sensitive Detection of HBsAg Mutants by a Gap Ligase Chain Reaction Assay

Bloodborne Pathogens and Hepatitis, National Microbiology Laboratory, Canadian Science Centre for Human and Animal Health, Winnipeg, Manitoba R3E 3P6, Canada

Received 27 November 2001/ Returned for modification 10 February 2002/ Accepted 21 April 2002

Hepatitis B virus (HBV) surface gene variants have been associated with diagnostic escape and immune escape following vaccination. The most common mutation observed in these variants is a glycine-to-arginine substitution at amino acid 145 (G145R). In order to sensitively detect the presence of this mutant in serum, a new molecular detection system was developed; in this new system, a gap ligase chain reaction (gLCR) assay was coupled with electrochemiluminescence detection of reaction products. The gLCR assay could detect approximately 10 copies of mutant DNA and could discriminate low levels of mutant DNA in the presence of excess wild-type DNA. Detection of the G145R mutant in clinical specimens was evaluated by testing 56 suspect serum specimens. The G145R mutation was observed in 18 of 28 HBV-DNA-positive samples. The approximate percentage of mutant present in each specimen was calculated by comparison with a standard curve of an increasing ratio of mutant DNA to wild-type DNA. Most samples contained a very low percentage of mutant virus (approximately 5%), with an observed range of approximately 3 to 74%. The G145R mutation was most frequently observed in specimens producing a diagnostic anomaly or from transplant patients but was also observed in specimens from vaccinated individuals and specimens in which HBsAg diagnostic escape was suspected. Therefore, the gLCR assay is a sensitive and specific method for detection of G145R mutants, which could be modified to include the detection of other HBV mutants.

This article has been cited by other articles:

• Osiowy, C., Gordon, D., Borlang, J., Giles, E., Villeneuve, J.-P. (2008). Hepatitis B virus genotype G epidemiology and co-infection with genotype A in Canada. J. Gen. Virol. 89: 3009-3015 [Abstract] [Full Text]  
• Osiowy, C., Giles, E., Tanaka, Y., Mizokami, M., Minuk, G. Y. (2006). Molecular Evolution of Hepatitis B Virus over 25 Years.. J. Virol. 80: 10307-10314 [Abstract] [Full Text]  
• Osiowy, C., Villeneuve, J.-P., Heathcote, E. J., Giles, E., Borlang, J. (2006). Detection of rtN236T and rtA181V/T Mutations Associated with Resistance to Adefovir Dipivoxil in Samples from Patients with Chronic Hepatitis B Virus Infection by the INNO-LiPA HBV DR Line Probe Assay (Version 2).. J. Clin. Microbiol. 44: 1994-1997 [Abstract] [Full Text]  
• Osiowy, C., Giles, E. (2003). Evaluation of the INNO-LiPA HBV Genotyping Assay for Determination of Hepatitis B Virus Genotype. J. Clin. Microbiol. 41: 5473-5477 [Abstract] [Full Text]