Multiplex-PCR for simultaneous detection and genotyping of hepatitis B virus in plasma and serum

Abstract

Hepatitis B virus (HBV) is the great medical problem worldwide in which 90% of acute HBV infection in adult is usually cleared, and 5-10% becomes chronic carrier. HBV could be classified into 8 genotypes (A-H). However, genotype B and C are predominantly reported in Asia. Currently, several studies revealed that clinical course and outcome of antiviral therapy were depended on the genotype. In this study, in-house multiplex-PCR was implemented for simultaneous detection and identification of HBV genotypes A, B, and C in serum and plasma. The pre-S1 to S gene was targeted for primers design. Of 100 HBsAg-positive and 40 HBsAg-negative plasma and serum were included in this study. Ninety of 100 HBsAg positive samples (90%) were positive and all of 40 HBsAg negative samples (100%) were negative by PCR, respectively. Among these positives, 7 (7.8%) and 75 (83.3%) were respectively identified to be genotype B and C. Three of these (3.3%) were mixed infection of genotype B and C and 5 (5.6%) were categorized into unclassified genotype. Apart from its role in epidemiological studies and treatment on HBV infection, multiplex-PCR method developed in this study provides a rapid and specific tool for detection and genotyping of HBV.