Molecular and Biochemical Study in Chronic Hepatitis B Patients in Anbar Province.

Abstract

Hepatitis B is a potentially life-threatening liver infection caused by a specific type of virus called hepatitis B virus (HBV). Iraq is among the intermediate HBV endemic countries because the infected rate of HBV is between (3%-4.5%) among the population and (2%-3%) among apparently healthy blood donors; therefore, this study was designed. A total of 40 patients with infectious phases of hepatitis B (HBeAg positive) divided into 20 female and 20 male and 40 non-infectious phases of hepatitis B patients (HBeAg negative) 18 female and 22 male and 40 healthy subjects 19 female and 21 male were included in this study during the period from February to September 2018. The study was carried out in the General Al-Ameria Hospital/ Al-Anbar province. Hepatitis B virus was identified as preliminary by Rapid immune chromatographic assay. Fully automated chemistry analyzer TOSOH, AIA 1800 ST, for the detection of HBsAg level was also done as a confirmatory test. Further, assay procedure for biochemical tests including Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), Alkaline phosphatase (ALP), Also total serum Bilirubin (TSB), Gamma-glutamyl transferase (GGT) and Total protein Albumin and Globulin were done by using automatic chemistry analyzer instrument (MNCHIP) for each serum specimen from patients and healthy individuals. On the other hand, measurement of non-enzymatic antioxidant which includes Vitamin C (V.C), and enzymatic antioxidant which includes superoxide dismutase (SOD). Further, the production of lipid peroxidation marker malondialdehyde (MDA) in addition interlukin-1 alpha (IL-1α), and vi interlukin-2 (IL-2) were also achieved by Enzyme- linked immunosorbent assay technique. Hepatitis B-DNA extraction was done by SaMag-12 automatic nucleic acid extraction system. Quantitative HBV-DNA was amplified using Real Time PCR. A total of 80 patients were revealed to be positive for HBsAg . Serum level of HBsAg was significantly increased in HBeAg positive patients compared to HBeAg negative patients (7779.9 ± 3898 vs. 3233.8 ± 2474 IU/ml) respectively. The viral load of HBV-DNA was also considerably increased in HBeAg positive patients compared to HBeAg negative patients (35328825 ±23101537 vs. 3115.1±1916.8 IU/ml). The chromatographic immunoassay was performed for all HBsAg positive patients to investigate the following markers: HBsAg, HBsAb, HBeAg, HBeAb, and HBcAb. In infectious form of hepatitis B, (those patients who were positive for HBeAg), Anti-HBsAg was not detected in any patients. Also, all patients (100%) were anti-HBs negative, in addition to that Anti-HBeAg was detected in 2.5% of patients while 97.5% of them were anti-HBeAg negative. Further, anti-HBc was detected in 67.5% of study patients. In biochemical and physiological aspect, the statistical analysis revealed that there was a significant increase in the mean level of serum AST, ALT, ALP in (HBeAg-positive) patients compared to (HBeAg negative) patients group and healthy control (P = 0.001). Regarding GGT levels increase in (HBeAg positive), and (HBeAg negative) patients group compared to healthy control (P value=0.002). Also, the level of total serum bilirubin, direct bilirubin, indirect bilirubin, total protein, albumin, and vii globulin showed no significant difference results in each HBeAg positive and HBeAg negative patients and healthy control. In oxidative stress markers; vitamin C levels in HBeAg-positive patients were lower than those observed in HBeAg-negative patients or in healthy volunteers (P value=0.001). However, SOD and MDA levels were significantly lower in both HBeAg-positive and HBeAg-negative patients than in the healthy volunteers. In immunological part, IL-1α levels appeared similar in the three groups of individuals. IL-2 levels were not significantly different between HBeAg-positive and control individuals but HBeAg-negative individuals demonstrated significantly higher levels than either HBeAg-positive patients or the controls (both, P = 0.001). The study concluded that qRT-PCR test have the superior clinical advantage among other techniques in quantitation viral load through study groups. Also, the biochemical parameters, especially ALT, AST, GGT, and ALP, were elevated in HBV patients significantly. The activities of vitamin C were significantly decreased in HBeAg positive patients while the activities of superoxide dismutase was significantly reduced in each HBeAg positive and HBeAg negative patients compared to healthy control. The activity of malondialdehyde has been markedly reduced in HBV patients. Further, Serum levels of IL-2 were decreased in HBeAg positive patients group relative to HBeAg negative patients.