Hepatitis b resistance testing and genotype determination

Sirwan Salman Sleman

Introduction: The aim of this study is to determine the predominant HBV genotypes for a number of samples and controls as well as to identify the mutant genes which thought to be responsible for the HBV antiviral drug resistance.

Methods: A number of samples (8 samples ), that were already collected from infected patients by the microbiology staff of the Manchester University hospital for the purpose of the study, are extracted and tested to determine the HBV genotypes and antiviral resistance. The techniques that are used for this purpose in this study are PCR amplification, gel electrophoresis and then sequencing of PCR products by using Applied Biosystems Invitrogen (ABI-3100).

Results: With gel electrophoresis, PCR sample products are compared with the intensity of ladder (marker), and the sequences are analysed by using Bioedit®  (Bioinformatics software) and then genotypes of the HBV are retrieved from the NCBI tool.

Conclusion: We found that the infected samples are predominantly returned to viral genotypes D as well as it is noticed that the resistance to the HBV antiviral drugs were related to the replacement of some amino acids in a viral gene by other wild mutant one ,especially at the amino acid codon 180 position there was a mutation where Methionine was replaced with mutant Leucine and also at the codon 204 where Methionine was replaced with mutant Valine.

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