Identification of a hepatitis B virus core promoter mutant by PCR- RFLP in patients suffering from chronic liver disease, Uttar Pradesh, India.
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.


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 (68.8%) of 16 patients, excluding 1 patients with mixed type mutation was detected in inactive HBsAg carriers, 39 (81.3%), excluding 2 patients with mixed type was detected in chronic hepatitis B, and 4/7 (57%) in patients with liver cirrhosis were found.


CONCLUSIONS
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.
Kumar
Ajay
A
Centre for Biotechnology, University of Allahabad, Allahabad, Uttar Pradesh, India. ajaykumarbiotech@rediffmail.com
Tiwari
Balesh Kumar
BK
Chaudhary
Ajay Kumar
AK
Pant
Sanjay
S
Narang
Sushil
S
D004279:DNA, Viral / Q000032:analysis*; D005260:Female; D006512:Hepatitis B Core Antigens / Q000235:genetics*; D006515:Hepatitis B virus / Q000235:genetics*; D019694:Hepatitis B, Chronic / Q000235:genetics*; D006801:Humans; D007194:India; D008297:Male; D009154:Mutation / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D011401:Promoter Regions, Genetic; D014758:Viral Core Proteins / Q000235:genetics*
D004279:DNA, Viral; D006512:Hepatitis B Core Antigens; D014758:Viral Core Proteins