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Detection of Intrahepatic Hepatitis B Virus DNA and Correlation with Hepatic Necroinflammation and Fibrosis

Danny Ka-Ho Wong,^1, Man-Fung Yuen,^1, Eric Tse,² HeJun Yuan,^1,3 Simon Siu-Man Sum,¹ Chee-Kin Hui,¹ and Ching-Lung Lai^1*

Division of Gastroenterology and Hepatology,¹ Division of Hematology and Oncology, Department of Medicine, The University of Hong Kong, Queen Mary Hospital, Hong Kong,² Department of Medicine, Fudan University, Zhongshan Hospital, Shanghai, People's Republic of China³

Received 23 June 2003/ Returned for modification 22 October 2003/ Accepted 27 March 2004

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 ß-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 ß-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 10⁶ versus 6.65 x 10⁶ 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.

D.K.-H.W. and M.-F.Y. contributed equally to the study.

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