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Genotyping of HBV using type specific primer PCR and restriction fragment length polymorphism-PCR

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Research Paper 01/03/2018
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Genotyping of HBV using type specific primer PCR and restriction fragment length polymorphism-PCR

Imran Riaz Malik, Sawera Nayyab, Ghulam Mujtaba, Qamar Bashir
Int. J. Biosci.12( 3), 194-200, March 2018.
Certificate: IJB 2018 [Generate Certificate]
Key: HBV: Hepatitis B VirusRFLP-PCR: Restriction Fragment Length polymorphism-PCRTSP-PCR: Type Specific Primer-PCR

Abstract

Globally, about 2.5 billion people are infected with HBV. In Pakistan, about seven million people are infected with hepatitis B. Hepatitis B is a liver inflammation caused by viral infection and can develop into liver carcinoma if not treated. HBV is categorized into eight molecular genotypes. The main objective of this study was to evaluate the efficacy of most effective method for genotyping as well as genotype distribution in Lahore, Pakistan. 130 HBV DNA positive samples were collected from different hospitals of Lahore, DNA extracted, and was preceded for amplification using type specific primer PCR and RFLP-PCR. Genotype specific amplicons were analyzed on 1.5% agarose gel electrophoresis. Results showed that genotype D was most abundant in Punjab, as first method (TSP-PCR) showed the 90.8% samples were of genotype D and 9.2% samples having the genotype C. Second method RFLP-PCR gave 94.6% samples were genotype D and 5.3% were genotype C after digesting by two enzymes Psu1 and Sty1. According to our sampling data genotyping comparison gave results: TSP-PCR showed that male have 93.50% genotype D and 6.50% genotype C and females have 88.60% genotype D and 11.40% genotype C. Second method RFLP-PCR showed that males have 96.10% genotype D and 3.90% genotype C and females have 92.40% genotype D and 7.60% genotype C. Statically analysis of both methods gave a significant P (> 0.5). It is concluded that both methods are very effective, sensitive and reliable techniques. Genotype D was found most dominant among Punjab population.

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