Application of PCR-LDR-nucleic acid detection strip in detection of YMDD mutation in hepatitis B patients treated with lamivudine.

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J Med Virol. 2010 Jul;82(7):1143-9.

Application of PCR-LDR-nucleic acid detection strip in detection of YMDD

mutation in hepatitis B patients treated with lamivudine.

Xu G, You Q, Pickerill S, Zhong H, Wang H, Shi J, Luo Y, You P, Kong H, Lu F, Hu

L.

Ustar Biotechnology (Hangzhou) Ltd, Zhejiang Province, People's Republic of

China.

Abstract

Chronic hepatitis B virus (CHBV) infection causes cirrhosis and hepatocellular

carcinoma. Lamivudine (LAM) has been successfully used to treat CHBV infections

but prolonged use leads to the emergence of drug-resistant variants. This is

primarily linked to a mutation in the tyrosine-methionine-aspartate-aspartate

(YMDD) motif of the HBV polymerase gene at position 204. Rapid diagnosis of

drug-resistant HBV is necessary for a prompt treatment response. Common

diagnostic methods such as sequencing and restriction fragment length

polymorphism (RFLP) analysis lack sensitivity and require significant

processing. The aim of this study was to demonstrate the usefulness of a novel

diagnostic method that combines polymerase chain reaction (PCR), ligase

detection reaction (LDR) and a nucleic acid detection strip (NADS) in detecting

site-specific mutations related to HBV LAM resistance. We compared this method

(PLNA) to direct sequencing and RFLP analysis in 50 clinical samples from HBV

infected patients. There was 90% concordance between all three results. PLNA

detected more samples containing mutant variants than both sequencing and RFLP

analysis and was more sensitive in detecting mixed variant populations. Plasmid

standards indicated that the sensitivity of PLNA is at or below 3,000 copies per

ml and that it can detect a minor variant at 5% of the total viral population.

This warrants its further development and suggests that the PLNA method could be

a useful tool in detecting LAM resistance. © 2010 Wiley-Liss, Inc.

PMID: 20513076 [PubMed - in process

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