SEROPROFILE, GENETIC DIVERSITY AND DRUG RESISTANCE OF HEPATITIS B VIRUS AMONG INFECTED INDIVIDUAL ATTAENDING MAMA LUCY KIBAKI
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Date
2016
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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