Detection of Hepatitis B Virus (HBV) DNA among Blood Donors with HBsAg Positive in Tuban District Based on Nested Polymerase Chain Reaction Technique (Nested PCR)

Abstract

Hepatitis B virus (HBV) infection is a health problem in the world including Indonesia. It is proven by increasing incident from year to year in detected cases of patients infected with HBV either acute, chronic or on liver cirrhosis, and mild HBV infection is often accidentally detected during blood tests. The blood donors in Tuban district only got serological examination for the detection of Hepatitis surface antigen (HBsAg) but have not been specifically got examination by Nested Polymerase Chain Reaction (Nested PCR) for the detection of HBV DNA. Examination of high number of samples by nested PCR could provide better result (positive). The purpose of this study is for the detection of HBV DNA by nested PCR technique in HBsAg positive blood donors in the Indonesian Red Cross (PMI) of Tuban District. This study is an experimental laboratory research. Blood samples from 13 HBsAg positive samples were obtained from 150 blood donors at Indonesian Red Cross (PMI) of Tuban District. The method used in this study was a nested PCR using primer pair 7 and 8 for first PCR also HBS1 and HBS2 for second PCR. Research was established in the biology laboratory of Universitas Ronggolawe (Unirow) Tuban and Institute of Tropical Disease, Universitas Airlangga (ITD-Unair) Surabaya from March to May 2012. Results showed out of 13 HBsAg positive samples there were 3 (3/13 = 23.08%) HBV DNA detected using primers P7 and P8 then 10 (10/13 = 76.92%) were detected using primers HBS1 and HBS2. Usage two primer pairs in nested PCR can detect all (100%) HBV DNA because the negative PCR results with a single primer pair can be detected with another primer pair.

Keywords: hepatitis B virus DNA, blood donors, Nested PCR