A Viral Kinetic Study Using Adefovir Dipivoxil Mono Therapy In Patients With Chronic Hepatitis B

AIM1. To study the viral kinetic of Adefovir Dipivoxil therapy of Chronic Hepatitis B infected patients 2.To study the changement of cccDNA levels during ADV thrapy on patients with Hepatitis B expecially after three months, which is the critical turing from the first period to the second period.METHEDSEstablish a new real-time fluorescent PCR assay to detect hepatitis B virus covalently closed circular DNA(HBV cccDNA).Plasmids carrying HBV fragment were ampli fied to establish the standard curve. In order to improve the specificity of the real-time PCR,we used the specific primer set and digested the relaxed circular DNA and line double strands DNA according to Plasmid SafeTM TP dependent Dnase.we also ver-ified the sensitivity and reproducibility of the method. Research on 13 chronic hepa-titis B patients,we amplified the real-time PCR to measure the HBV DNA and ccc-DNA levels,combination with detecting sera HBV DNA,HBsAg and HBeAg levels to study the virus kinetic both in liver and sera.RESULTWe successfully established a real-time fluorescent PCR assay for cccDNA with the linear range from 102 to 108 copies per millilite.Patients'liver and serum cccDNA positive ratiowere 100% and 35.5%. Intrahepatic HBV DNA levels correlatied posi-tively with sera HBV DNA (r=0.823, P<0.001),HBsAg (r=0.623, P=0.002) and HBeAg (r=0.507, P=0.023). Intrahepatic cccDNA levels correlatied positively with sera HBV DNA (r=0.499, P=0.025),HBsAg (r=0.546, P=0.013),it doesn't have correlation with HBeAg. After oraling the adv 12 weeks,Those patients'intrahepatic HBV DNA levels and cccDNA levels declined rapidly, reduced by 95.17%,96.63% respectively, which is parallel to sera HBV(99.6%). CONCLUSIONS1.Established a real-time fluorescent PCR assay for cccDNA 2. Intrahepatic HBVDNA levels correlatied positively with sera HBVDNA,HBsAg and HBeAg. Intrahepatic cccDNA levels correlatied positively with sera HBVDNA,HBsAg.3.Due to 12 wee- ks'adefovir dipivoxil therapy, Patients'intrahepatic HBVDNA levels and cccDNA levels declined rapidly, reduced by 95.17%,96.63% respectively, which is parallel to sera HBV(99.6%).