Seroprofile, genetic diversity and drug resistance of hepatitis b virus among HIV infected individuals attending Mama Lucy Kibaki comprehensive care clinic in Nairobi, Kenya

Abstract

Human immune deficiency virus (HIV) and Hepatitis B virus(HBV) coinfection is highly prevalent among high risk populations including pregnant women and infants. This poses a global public health challenge in laboratory diagnosis and is a major consideration for anti-HIV treatment. These viruses share common modes of transmission that is; through blood and body fluids. Further, there is little information on sero-profiles and circulating HBV genotypes in Kenya. This study aimed at determining seroprofiles, genetic diversity and drug resistance of HBV among HIV infected individuals attending comprehensive care clinic of Mama Lucy Kibaki Hospital Nairobi, Kenya. Ethical approval was sought from Kenyatta university ethics review committee and a cross-sectional study was conducted whereby the participants/guardians who gave consent/assent were included into the study. Their demographic data was collected using a questionnaire and 5ml of blood was collected from each participant using systematic sampling technique. The HBV seroprofiles were determined using the HBV-5 panel rapid diagnostic cassette according to the manufacturer’s instructions (Healthaw Medical limited, Hangzhou, China) . Viral DNA was extracted using Qiagen® Miniviral DNA isolation kit and the HBV-pol gene amplified by nested PCR. The amplified products were sequenced using the Big Dye® sequence terminator kit (Applied Biosystem®) on an automated ABI 310 sequencer (Applied Biosystem, Foster City CA). The generated sequences of HBV were analysed for drug resistance and genetic diversity determined using Molecular Evolutionary Genetics Analysis (MEGA5). Four hundred participants were recruited and 293 were females, 107 were males with their age ranging between 4 months and 73 years. Of the 400 sera; (111) 27.8% were HBV immunized, 19 (4.8%) were recovery cases, 12 (3%) had acute disease, 10 (2.5%) were chronic, 9 (2.3%) had occult HBV and 7 (1.8%) asymptomatic. The prevalence of HBV/HIV was found to be 7.25% based on the presence of surface antigen. After the confirmation of HBV DNA by gel electrophoresis, 13 samples were successfully amplified, purified and sequenced. All the 13 sequences were confirmed as HBV genotype A. Nucleotide drug resistance mutations were found in six (6) participants’ samples. These were rtV173L, rtL180M, rtM204V which are major mutations associated with lamivudine, telbivudine and emtricitabine resistance. This study indicates that the utility of HBV seromarkers and infection staging are important in disease diagnosis. The findings confirm that, HBV genotype A remains the most predominant genotype circulating in Nairobi.This study proposes a need for a continuous surveillance of HBV genotype trends and evolution of drug resistance because the current findings have major implications on treatment of HBV in Kenya.