Determination of serum hepatitis B virus DNA in HBV endemic region: Clinical significance and correlation with serological markers, ALT and AST

Abstract

Objective: The aim of this study is to investigate, demographics and the relationship between serologic markers, liver enzymes and HBV DNA levels from the patients whose samples were sent to microbiology laboratory for HBV DNA polymerase chain reaction (PCR) test, in one of the greatest cities of Southeast Anatolia, where HBV infection is extremely common. Method: Between October 2009 and October 2010, blood samples of 1662 patients were sent to our laboratory for HBV DNA PCR testing as part of a routine examination for viral hepatitis in Sanliurfa Training and Research Hospital in Turkey. Hepatitis markers (HBsAg, HBeAg, anti-HBs, anti-HBc IgM) and liver function enzymes (ALT, AST) are tested simultaneously in these samples. HBsAg, HBeAg, anti-HBs and anti- HBc IgM levels were assessed with chemiluminescent microparticle immunoassay method (Architect-i2000 SR, Abbott, Ireland). ALT and AST levels were measured with commercial kits in Roche Modular-P and Roche- Integra autoanalysator (Roche Diagnostics Mannheim, Germany). HBV DNA PCR: Isolation of HBV DNA from serum samples was done with M24sp automated extraction device (Abbott, USA). Purified DNA was amplified with Real-Time PCR method with the help of Abbott-m2000 sample preparation system (Abbott, USA). Results: The mean age was 33.7 (Min-Max: 0-89) years old and the majority of the patients were male (Male/Female: 1103/559) in the study. HBsAg was positive in 96.2% of the patients and the median ALT level of these patients were 94 u/l (Min-Max: 4-2390 u/l), while median AST level was 74 u/l (Min-Max: 7-2505 u/l). HBV DNA was positive in 94.6% of HBsAg positive patients and the median HBV DNA level was 3.9x104 copies/ml (Min-Max: 35-3.4x109 copies/ml). HBeAg was positive in 17.1% of HBsAg positive patients. Median levels of ALT and AST in HBsAg negative patients (3.8%) were 32 u/l (Min-Max: 7-497 u/l) and 33 u/l (Min- Max: 13-280 u/l) respectively. HBV DNA was positive in only 7.9% of the patients with median HBV DNA level of 155 copies/ml (Min-Max: 34-624 copies/ml). HBeAg was found negative in 71.4% of patients with negative HBsAg and positive HBV DNA. In 26 of 1662 patients (1.6%), both HBsAg and Anti-HBs were negative (N/N). Only two of the N/N patients (7.7%) were found anti-HBc positive. Mean ALT and AST levels of two anti-HBc positive patients were 29.5 u/l and 30.5 u/l, respectively. HBV DNA was negative in one of these patients, and the other was found positive with a level of 3×102 copies/ml. Conclusion: To acquire more reliable results on the diagnosis and follow-up of the treatment of Hepatitis B, quantitative HBV DNA tests should be used in together with more frequently used serologic and biochemical methods.