Title: Sensitiviy of pyrosequencing and PCR product direct sequencing in detection of drug resistance genes in hepatitis B virus:a comparative study
Abstract: OBJECTIVE To evaluate the potential value of pyrosequencing method in the detection of HBV resistance genes through the comparison with PCR product direct sequencing method.METHODS The patients with chronic hepatitis B treated with lamivudine were selected,the sensitivity and specificity of two methods mentioned above were compared by detecting the variation of lamivudine resistance mutation site rt180aa.RESULTS Both pyrosequencing and Sanger sequencing could detect the dominant virus(90%) exactly.When the proportion of rt180 mutation ranged from 90% to 50%,which was detected by pyrosequencing,the detection value by Sanger sequencing were reduced to 76.92% and 50.00%;when the proportion value of rt180M was within 50%-10% determined by pyrosequencing,it would be reduced to 10.00%-15.38% by Sanger sequencing method;that means quasispecies less than 10% could be only detected by pyrosequencing rather than by Sanger sequencing.CONCLUSION As compared with the Sanger sequencing,pyrosequencing method is with better sensitivity in the detection of variation of hepatitis B virus,which can detect the quasispecies and its population drift during the antiviral therapy process,with great significance in research on the antiviral effect and the forecast of drug resistance.
Publication Year: 2012
Publication Date: 2012-01-01
Language: en
Type: article
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