Pyrosequencing for detection of drug resistant relevant mutation in the polymerase gene of Hepatitis B virus and its clinical application / 中华检验医学杂志

ABSTRACT

Objective To explore the accuracy and clinical application of Pyrosequencing for detection of drug resistant relevant mutation in the polymerase gene of Hepatitis B virus (HBV).Methods Compared with Sanger sequencing,the accuracy and sensitivity of Pyrosequencing were assessed.Pyrosequencing was used to determine the serum of 1164 patients with chronic Hepatitis B and its results were analyzed.Results The sensitivity of Pyrosequencing was 1 × 103 KIU/L,the same as Sanger sequencing.But Pyrosequencing was more stable and specific than Sanger sequencing to detect low rate of mutation with over 10％ mutation.The mutations couldn't be detected in 248 patients with chronic Hepatitis B,but positive results occurred in 919 patients,among them,61.5 ％ without mutation,38.5 ％ with mutation (including 47.5％ of the single locus mutation and 52.5％ of the combined mutation).Other sites had different degree of mutations except the sites of rtI169T and rtA194T; the main mutation sites in patients with chronic Hepatitis B were rtM204V/I (32.1％),rtL180M (19.8％),rtA181V/T (6.7％) and rtN236T (5.3％),in which fractional mutation had high ratio in the sites of rtA181V/T (6.7％) and rtN236T (5.3％).Conclusions Pyrosequencing has good sensitivity,specificity and accuracy and can early detect the drug resistant relevant mutation in the polymerase gene of HBV,which is suitable for monitoring patients with chronic Hepatitis B for the treatment of nucleoside analogues (acid).The different sites and frequencies of mutations may be associated with different habits of doctors for using different anti-HBV drugs.