Detection of Hepatitis B Virus DNA among Abdominal Typhus Patients with Hepatitis B Virus Co-Infection in Tuban District Based on Nested PCR Technique

Abstract

Typhoid fever is often known by the name of abdominal typhus that may cause liver disruption so that arise disease complication which was known as typhoid hepatitis. Detection of abdominal typhus generally using routine blood test techniques that based on antigen and antibody reactions which is take a long time to obtain positive results so that these test cannot diagnose quickly and accurately. The detection of this disease in Tuban district is only using simple routine blood tests (eg Widal), urine and feces; it is difficult to diagnose the disease if there is complication. Abdominal typhus patients in Tuban district never got a specific examination technique by Nested Polymerase Chain Reaction (Nested PCR) to detect the occurrence of hepatitis B virus co-infection.

The purpose of this study to detect hepatitis B virus DNA among abdominal typhus patients with hepatitis B virus co-infection by using Nested PCR techniques.

This study is an experimental laboratory research. Blood samples from 9 positive HBsAg samples, which was obtained from 30 abdominal typhus patients in R. Kusma Hospital of Tuban District. The method used in this study was a nested PCR using primers P1 and P2 for the first PCR also primers HBV1 and HBV2 for the second PCR. Research carried out at the Biology Laboratory of Universitas  Ronggolawe (Unirow) Tuban and Institute of Tropical Disease, Universitas Airlangga (ITD Unair) in Surabaya, from March to May 2009.

The results showed out of 9 positive HBsAg samples there were 4 (44.44%) HBV DNA detected using primer pair P1 and P2 then 1 (11.11%) were detected using primer pair HBS1 and HBS2. The use of two pairs of primers in nested PCR can detect more HBV DNA, because of the negative PCR results using a single primer pair can be detected using another primer pair.

Conclusion of the study is usage of nested PCR technique using two different primer pairs can detect more HBV DNA in abdominal typhus patients with hepatitis B virus co-infection as much as 55.55%.

Keywords: hepatitis B virus DNA, abdominal typhus patients, Nested PCR