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Journal of Clinical Microbiology, January 1999, p. 161-164, Vol. 37, No. 1
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Nucleic Acid-Based Cross-Linking Assay for Detection and Quantification of Hepatitis B Virus DNA

Vicky C. H. Lai,1 Richard Guan,2 Michael L. Wood,3,* Su Kong Lo,4 Man-Fung Yuen,1 and Ching-Lung Lai1

Department of Medicine, University of Hong Kong, Hong Kong1; Medical Clinic One2 and Department of Medicine, National University of Singapore,4 Singapore; and NAXCOR, Menlo Park, California3

Received 7 July 1998/Returned for modification 7 August 1998/Accepted 13 October 1998

A nucleic acid photo-cross-linking technology was used to develop a direct assay for the quantification of hepatitis B virus (HBV) DNA levels in serum. Cross-linker-modified DNA probes complementary to the viral genomes of the major HBV subtypes were synthesized and used in an assay that could be completed in less than 6 h. The quantification range of the assay, as determined by testing serial dilutions of Eurohep HBV reference standards and cloned HBV DNA, was 5 × 105 to 3 × 109 molecules of HBV DNA/ml of serum. Within-run and between-run coefficients of variation (CVs) for the assay were 4.3 and 4.0%, respectively. The assay was used to determine HBV DNA levels in 302 serum samples, and the results were compared to those obtained after testing the same samples with the Chiron branched-DNA (bDNA) assay for HBV DNA. Of the samples tested, 218 were positive for HBV DNA by both assays and 72 gave results below the cutoff for both assays. Of the remaining 12 samples, 10 were positive for HBV DNA by the cross-linking assay only; the 2 other samples were positive by the bDNA assay only. Twenty-eight samples had to be retested by the bDNA assay (CV, >20% between the results obtained from the testing of each sample in duplicate), whereas only three samples required retesting by the cross-linking assay. The correlation between the HBV DNA levels, as measured by the two tests, was very high (r = 0.902; P = 0.01). We conclude that the cross-linking assay is a sensitive and reproducible method for the detection and quantification of HBV DNA levels in serum.

* Corresponding author. Mailing address: NAXCOR, 4600 Bohannon Dr., Suite 220, Menlo Park, CA 94025. Phone and Fax: (650) 328-3393. E-mail: mikelwood{at}aol.com.

Journal of Clinical Microbiology, January 1999, p. 161-164, Vol. 37, No. 1
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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