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Quantitation of covalently closed circular hepatitis B virus DNA in chronic hepatitis B patients
Author(s) -
Wong Danny KaHo,
Yuen ManFung,
Yuan HeJun,
Sum Simon SiuMan,
Hui CheeKin,
Hall Jeff,
Lai ChingLung
Publication year - 2004
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.20353
Subject(s) - cccdna , hepatitis b virus , hbeag , hepatology , circular dna , virology , hepatitis b , liver biopsy , medicine , virus , biology , biopsy , hbsag , gene , biochemistry , genome
This study examined a signal amplification assay, the Invader assay, for the quantitation of hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) in liver biopsies and sera. DNA was extracted from liver biopsy and serum samples were collected from 16 hepatitis B e antigen (HBeAg)‐positive and 36 antibody‐to‐HBeAg‐positive (anti–HBe‐positive) chronic hepatitis B patients. The amount of total HBV DNA and cccDNA was measured using the Invader assay. Anti–HBe‐positive patients had lower median total intrahepatic HBV DNA ( P < .001) and intrahepatic cccDNA levels ( P = .001) than HBeAg‐positive patients. Intrahepatic cccDNA correlated positively with the total intrahepatic HBV DNA ( r = 0.950, P < .001). However, the proportion of intrahepatic HBV DNA in the form of cccDNA was inversely related to the amount of total intrahepatic HBV DNA ( r = −0.822, P < .001). A small amount of cccDNA was detected in 39 of 52 (75%) serum samples. Anti‐HBe‐positive patients had lower median serum cccDNA levels than HBeAg‐positive patients ( P = .002). Serum HBV DNA correlated positively with intrahepatic total HBV DNA ( r = 0.778, P < .001) and intrahepatic cccDNA ( r = 0.481, P = .002). In conclusion , the Invader assay is a reliable assay for the quantitation of cccDNA. Serum and intrahepatic total HBV DNA and cccDNA levels become lower as the disease progresses from HBeAg‐positive to anti–HBe‐positive phase, with cccDNA becoming the predominant form of intrahepatic HBV DNA. (H EPATOLOGY 2004;40:727–737.)