Relationships Of CccDNA In HBV Infeced Subjecs' Sera With Virus Replication And Liver Damage

Objects: to preparily assess the clinical value of hepatitis B virus (HBV) covalently closedcircle DNA(cccDNA) in HBV infeced subjecs' sera for the predictition virus reactivation and liver histology damage.Methods: plasma samples of 35 subjects with no-HBV infected hepatitis and of 663 subjects with HBV infected disease including chronic inactive HBsAg cariers (sAg-AsC~*), chronic asympathetic HBV carriers (V-AsC~*), acute hepatitis B (AHB), chronic hepatitis (CHB), liver cirrhosis(LC) and hepatocellular carcinoma (HCC) 35,70,41,392,65,60 respectively, were detected with polymerase chain reaction and molecular-probes hybridization. PCR-molecular-beacon technique, which is capable of differentiation of HBV viral genomic DNA and cccDNA as HBV viral DNA has a gap in the minus-strand and an incomplete plus-strand of variable length that we is able to design selective cccDNA primers to amplify the region corresponding to the gap and incomplete region in the partially double-strand virion DNA. quantification of HBVDNA, HBV-marker, liver function indexes in serum were tested in realtime PCR, ELISA and rutin biochemical methods respectively. The inflammation grade of liver biopsies from patients with ALT＜2ULN and HBVDNA positivity were assessed, and the cccDNA in serum was deteced and the ALT level of some Some HBV carriers with sero cccDNA positivity/negativity were followed for 1 month.Results: The diference of cccDNA positive ratio in HBeAg positive group (144/446, 32.2%) Vs that in HBeAg negative group 12/217, 5.5%) was significant (χ~2=58.08, P＜0.01). The 156 sero cccDNA positive samples all were in the 511 plasma samples of HBVDNA positive and the sero cccDNA were all negative in all 152 HBVDNA negative samples (χ~2=60.68, P＜0.01). Of 663 HBsAg positive subjects there were 95 serum alanine aminotrasferase normal in 507serum cccDNA negative group and 15 serum alanine aminotrasferase normal in 156cccDNA positive group(χ~2=7.17, P＜0.05) but the serum bilirubin abnormal ratio was of no-significance (35/156Vs109/507,χ~2=0.52, P＞0.05). Of 181 with ALT＜80U/L(including 79 HBVcarriers) and 330 ALT＞80U/L(mAST＜2ULN141 and mAST＞2ULN 189) in all 511 patients with serum HBVDNA positive, the cccDNA positive ratio correlated with serum ALT (21/181Vs135/330,χ~2=47.34, P＜0.01) but din't with mAST (61/80Vs74/115,χ~2=0.56, P＞0.05). The degree of necroinflammation of the liver biopsies from 16 HBV carriers (including 6 sero cccDNA positive Ssubjects with G1, G2, G3 1, 3, 2 respectively) and from 34 chronic hepatitis B with ALT＜80U/L revealed that serum cccDNA was correlated with necroinflammation activenes (χ~2=35.38, P＜0.01). The ALT of all 18 with sero cccDNA positivity and of 30 with that negativity flared in 48 HBV carriers followed for 1 month (χ~2=40.2, P＜0.01).Conclusin: Serum cccDNA may be a serum marker indicating HBV replication and liver damage.