Molecular inversion probe-based single base mutations of hepatitis B virus resistance gene analysis

Abstract

Objective To establish a molecular inversion probe（MIP）method for detection of single base drug-resistance mutation in Hepatitis B virus（HBV）gene. Methods The HBV wild type and YVDD mutant strain were isolated by Daping Hospital of the Third Military Medical University.The MIP was designed and applied to detect the HBV drug-resistance YVDD mutation in one case of wild type and one case of YVDD mutant HBV strain isolated previously.The results of MIP method were compared with that of sequencing to evaluate the detection accuracy. Results Thermal cycling single-base extension and connection reaction performed by Taq DNA Ligase and Ampligase DNA Ligase could ensure the specificity of the detection.The optimum probe concentration of MIP was 1 nmol/L.Through detection of the target gene with different DNA concentrations, the detection sensitivity of MIP was determined as 1 nmol/L. The results of MIP were consistent with that of sequencing method in detection of the clinical samples. Conclusion MIP is successfully used to detect single-base drug-resistance mutation in HBV gene.（Chin J Lab Med，2014,37：337-341） Key words: Hepatitis B virus; Drug resistance, viral; Molecular probe techniques; Genotype; Polymorphism, single nucleotide