Detection of intrahepatic hepatitis B virus DNA and correlation with hepatic necroinflammation and fibrosis.
Assessment of intrahepatic hepatitis B virus (HBV) DNA levels in patients with chronic hepatitis B is important in understanding the natural history of the disease and designing antiviral therapy regimens. However, there is no standardized method for the measurement of intrahepatic HBV DNA levels. We describe a convenient novel method for the measurement of intrahepatic HBV DNA levels based on a modified COBAS Amplicor HBV Monitor test for HBV DNA measurement and real-time PCR beta-actin gene detection for human genomic DNA (hgDNA) quantitation. Fifteen hepatitis B e antigen (HBeAg)-positive patients, 26 patients positive for antibody to HBeAg (anti-HBe), and 8 control patients were recruited. The mean between-run coefficient of variation for the beta-actin real-time PCR assay was 15.4%. All eight control patients had undetectable intrahepatic and serum HBV DNA levels. All chronic hepatitis B patients had detectable intrahepatic HBV DNA levels, and all but one anti-HBe-positive patient had detectable serum HBV DNA levels. HBeAg-positive patients had higher median intrahepatic and serum HBV DNA levels than anti-HBe-positive patients (6,950 versus 676 HBV DNA copies/ng of hgDNA, respectively [P < 0.001] and 184 x 10(6) versus 6.65 x 10(6) copies/ml, respectively [P < 0.001]). The intrahepatic HBV DNA levels correlated strongly with the serum HBV DNA levels (r = 0.842; P < 0.001) and with the degree of fibrosis (P = 0.014). We conclude that the method that we describe is reliable and convenient for the measurement of intrahepatic HBV DNA levels and has potential clinical significance.
Wong
Danny Ka-Ho
DK
Division of Gastroenterology and Hepatology, Department of Medicine, The University of Hong Kong, Queen Mary Hospital, Pokfulam Rd., Hong Kong, China.
Yuen
Man-Fung
MF
Tse
Eric
E
Yuan
HeJun
H
Sum
Simon Siu-Man
SS
Hui
Chee-Kin
CK
Lai
Ching-Lung
CL
D000199:Actins / Q000235:genetics; D000328:Adult; D001483:Base Sequence; D017931:DNA Primers; D004279:DNA, Viral / Q000097:blood / Q000302:isolation & purification*; D005260:Female; D006509:Hepatitis B / Q000473:pathology*; D006513:Hepatitis B e Antigens / Q000097:blood; D006515:Hepatitis B virus / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007249:Inflammation / Q000473:pathology / Q000821:virology; D008099:Liver / Q000473:pathology* / Q000821:virology*; D008103:Liver Cirrhosis / Q000473:pathology / Q000821:virology; D008297:Male; D016133:Polymerase Chain Reaction
D000199:Actins; D017931:DNA Primers; D004279:DNA, Viral; D006513:Hepatitis B e Antigens