Detection of Hepatitis B Virus (HBV) Genotypes by Nested Polymerase Chain Reaction (PCR) Technique in Diyala Province

Abstract

Hepatitis B virus (HBV) infection is a public health concern because it causes liver diseases such as hepatocellular carcinoma and liver cirrhosis. Depending on the virus sequence homogeneity, ten HBV genotypes (A-J) were identified. The aim of this study is the molecular and serological detection of HBV utilizing the nested polymerase chain reaction (PCR) technique, and biochemical test for liver function to estimate the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and Alkaline phosphatase (ALP). A cross-sectional study was conducted in170 reviewers (103 males and 67 females) who attended the Baquba Teaching Hospital, Teaching laboratories, the Main Blood Bank, and Baladrooz General Hospital. Age ranging 13-75 years. The samples were collected from Diyala Province of Iraq, Enzyme- Linked Immune Sorbent Assay test (ELISA) was used to detect hepatitis B surface antigen (HBsAg) in serum. Automated machine (Mindray Bs-240 system) for quantitative determination of (ALT, AST, ALP) was used. Conventional Polymerase Chain Reaction (PCR) of HBV p gene was performed for HBV- DNA detection from serum samples, and Nested PCR technique with specific primers was used to detect HBV genotypes. Results of ELISA showed that 70 (41.20%) samples were positive and 100 (58.80%) samples were negative. PCR results showed that 14 (8.20%) samples were positive for HBV- DNA. Statistically, the positivity of HBV detected by ELISA was higher (41.2%) than positivity of HBV detected by PCR (8.20%). Liver function test found the levels of ALT (36.31±12.00), AST (47.92±21.00) ALP (100.24±45.92) was high in patients than controls. ALP and AST scored the highest sensitivity than ALT, while based on specificity, ALT and ALP scored the highest specificity than AST.