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Journal of Clinical Microbiology, February 2007, p. 553-558, Vol. 45, No. 2
0095-1137/07/$08.00+0     doi:10.1128/JCM.00709-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

A Genotype-Independent Real-Time PCR Assay for Quantification of Hepatitis B Virus DNA{triangledown}

Ying Liu, Munira Hussain, Stephen Wong, Scott K. Fung, Hyung Joon Yim, and Anna S. F. Lok*

Division of Gastroenterology, University of Michigan Medical Center, Ann Arbor, Michigan

Received 4 April 2006/ Returned for modification 14 June 2006/ Accepted 22 November 2006

Accurate quantification of hepatitis B virus (HBV) DNA levels is important for monitoring patients with chronic HBV infection and for assessing their responses to antiviral therapy. This study aimed to develop a real-time PCR assay that is sensitive and can accurately quantify a wide range of HBV DNA levels across the known HBV genotypes. An "in-house" real-time PCR assay using primers and a TaqMan probe in a highly conserved region of the HBV surface gene was designed. The assay was standardized against a WHO standard and validated against plasmids of HBV genotypes A through H. The linear quantification range was approximately 5 x 100 to 2.0 x 109 IU/ml. Results of samples from patients infected with HBV genotypes A through H tested using our real-time "in-house" PCR assay showed an excellent correlation with those of the Cobas Amplicor HBV Monitor (R2 = 0.9435) and the Cobas TaqMan HBV (R2 = 0.9873) tests. We have established a real-time PCR assay that is genotype independent and can accurately quantify a wide range of HBV DNA levels. Further studies of additional samples are ongoing to validate the genotype independence of our assay.

* Corresponding author. Mailing address: Division of Gastroenterology, University of Michigan Medical Center, 1500 East Medical Center Dr., 3912 Taubman Center, Box 0362, Ann Arbor, MI 48109-0362. Phone: (734) 615-4628. Fax: (734) 936-7392. E-mail: aslok{at}umich.edu.

{triangledown} Published ahead of print on 20 December 2006.

Journal of Clinical Microbiology, February 2007, p. 553-558, Vol. 45, No. 2
0095-1137/07/$08.00+0     doi:10.1128/JCM.00709-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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