A novel accurate amplification created restriction site method for determination of the wild type and the precore mutant hepatitis B virus (HBV) variants

The most common occurring hepatits B virus (HBV) mutation is the G to A mutation at nucleotide 1896 in the precore region. The aim of this study is to develop a novel accurate amplification created restriction site (ACRS) method for determination of the TGG wild type and the TAG precore mutant HBV variants. Two conserved and consensus specific and diagnostic primers introducing BstX I and Xag I cleavage site were designed in order to determine the G1896 wild type and the A1896 precore mutant HBV variants in all HBV genotype.The results of ACRS method were compared with sequencing data. In the ACRS method, three different informative patterns could be distingished for determination of the wild type, the precore mutant and mixed infection HBV variants. The results of ACRS method on thirty HBV isolates revealed the TAG precore mutant in 50% (15/30), the TGG wild type variant in 30% (9/30) and the mixed infection in 20% (6/30). The sequencing data of these samples were in agreement with the ACRS results. The developed ACRS method is a rapid and cost effective technique for detection of both the TGG wild type and the TAG HBV precore mutant variants. It can be performed for follow up of G1896A precore mutant variant in hepatits B virus infected subjects at routine molecular diagostic laboratories.

کلیدواژه ها:

Hepatitis B virus ، Precore mutant ، ACRS method ، Iranian patients