Objective and Aim: There was a paradigm shift of hepatitis B (CHB) diagnosis as clinicians are shifted to molecular diagnostic methods from serological one. Specially in molecular system to determine response of treatment as well as different stages of infection as well as recovery by quantification of viral DNA load through real time polymerase chain reaction (RT-PCR). The main objective of the study is to determine the various clinical stages of chronic hepatitis B through measuring virus load DNA by real-time polymerase chain reaction (RT-PCR)
Material and Methods: This is a retrospective study of those patients whose ALT (elevated) and HBeAg (positive) status is known. Serum fraction were initially obtained after 4 hour centrifugation of blood sample and nucleic acid was extracted at -80 °C. Qiagen DNA extraction kit were used to extract DNA. 48-well MiniOpticon by Bio-red machine and with the help of Geno-sense HBV quantitative PCR kit, real-time polymerase chain reaction (RT-PCR) was conducted.
Result: The study was conducted in 64 patients. It has been found that among this patients inactive carriers that is ALT normal and HBeAg-negative were 27 (42.2%) and rest of the patients had HBeAg-positive or HBeAg-negative with ALT elevated that is they were chronic active hepatitis B patients. HBeAg was negative in 42 (65.6%) and positive in 22 (33.4%) subjects. 15 (23%) patients were infected with Chronic hepatitis B among the patients who were HBeAg-negative. Among 64 subjects, detectable viral load was found in 55 (86%) CHB patients. A significantly lower (median 5.6 × 105) serum HBV DNA load were found in HBeAg-negative16 patients as compare to 26 patients with higher viral load (median 2.5 × 108) and were HBeAg-positive. It has also found that viral load was quite higher (median 1.5 × 103) in 27 inactive carriers. Antiviral therapy was started in HBeAg-negative 6 patients and HBeAg-positive 13 patients based on the viral load.
Conclusion: Stages of CHB can be determined by Quantitation of HBV DNA based on ALT (elevated or not) and HBeAg (positive or negative) status. For those patients who are inactive carriers and HBeAg-negative with respect to viral load it could play an important role in assessment and to decide on antiviral therapy.
Serological Markers Hbsag and Hbeag in Chronic Hepatitis B Carriers and their Correlation with Viral DNA by Polymerase Chain Reaction
