Asian Pacific Journal of Cancer Prevention, Vol 10, 2009 1173 HBV Core Promoter Mutant in Indian Patients Suffering from Chronic Liver Disease Asian Pacific J Cancer Prev, 10, 1173-1175 Introduction Hepatitis B virus (HBV) is a partially double-stranded DNA virus, which is a major causative agent of chronic, acute hepatitis, liver cirrhosis and hepatocellular carcinoma in Asian, African and Southern European countries. An estimated 350 to 400 million carriers are found around the world (Wang et al., 2005) and 1 to 2 million people have died from this virus (Mahoney, 1999). In India the prevalence rate of hepatitis is reported to vary from 1 to 13 percent; with an average of 4.7 percent (Thyagarajan et al., 1996). Hepatitis B envelope antigen (HBeAg) is considered as a marker for viral replication but seroconversion from HBeAg to anti-HBeAg positive generally indicates low level of viral production and low level of serum HBV DNA. This generally correlates with improvement of liver disease. HBV has a mutation rate 10-fold higher than other DNA viruses (Tong et al., 2005). In the natural course of chronic infection, loss of HBeAg expression and appearance of antibodies (anti-HBeAg) are usually accompanied by cessation of viral replication. The HBV variants appear during HBeAg seroconversion and they 1 Centre for Biotechnology, University of Allahabad, Allahabad, 2 Department of Gastroenterology & Hepatology, MLN Medical College, Allahabad, Uttar Pradesh, India *For Correspondence: ajaykumarbiotech@rediffmail.com Abstract Objective: The present study was designed to identify a core promotor mutation in the HBV genome in patients suffering from HBV related chronic liver disease. Materials and Methods: 154 chronic liver disease patients were selected for study of DNA extracted using a pure viral DNA extraction kit. The core promoter mutation was detected by the polymerase chain reaction- based restriction fragment length polymorphism (PCR- RFLP) method, using the Sau 3AI restriction enzyme to see if cleavage would occur at this specific site. Results: Among the total of 154, 78 patients were found positive for HBsAg and 71 samples were found to be positive for HBV DNA by first round PCR. The over all prevalence of core mutant was 51(71%) in the 71 patients. 11 (68.75%) of 16 patients, excluding 1 patients with mixed type mutation was detected in inactive HBsAg carriers, 39 (81.25%), excluding 2 patients with mixed type was detected in chronic hepatitis B, and 4/7 (57%) in patients with liver cirrhosis were found. Conclusion: Our study concluded that the prevalence of the core promoter mutation in the BCP region was higher in the patients with chronic hepatitis B than in liver cirrhosis and HBsAg carriers. The Sau3AI assay, which is much more convenient than sequencing, was shown to be useful for the detection of the core promoter mutant in an extensive number of clinical samples. Monitoring and detection of HBV variants by PCR-RFLP in chronic infection may improve the management of these patients Key Words: HBeAg - BCP - PCR-RFLP- core promoter mutant bring mutations in the precore region (PC) that prevent HBeAg synthesis. The most common of these mutations is a G to A substitution at nucleotide 1896, which prevents the production of HBeAg by introducing a premature stop codon into the open reading frame (ORF) of the PC. This mutation converts codon 28 of the pre core sequence to a termination codon (TGG-TAG) and therefore it prevents expression of HBeAg (Lok et al., 1994). Another common HBeAg variant is the basal core promoter mutant (BCP) characterized by point mutation in the promoter of both HBeAg mRNA and core protein mRNA .The most frequent core promoter mutation is the double A1762T and G1764A nucleotide exchange, which results in a substantial decrease in HbeAg expression but enhanced viral genome replication (Tong et al., 2005). As opposed to precore variants, core promoter mutations can be detected in patients who are either HBeAg +ve or –ve but most frequently found in the HBsAg +ve and HBeAg –ve/ anti-HBeAg +ve. The prevalence of the core promoter variant is about 40%, evenly distributed among the major HBV genotypes. The present study aimed to identify a hepatitis B virus core promoter mutation in patients suffering from chronic liver disease. RESEARCH COMMUNICATION Identification of a Hepatitis B Virus Core Promoter Mutant by PCR- RFLP in Patients Suffering from Chronic Liver Disease, Uttar Pradesh, India Ajay Kumar 1 *, Balesh Kumar Tiwari 1 , Ajay Kumar Chaudhary 1 , Sanjay Pant 2 , Sushil Narang 2