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Journal of Clinical Microbiology, August 2003, p. 3699-3705, Vol. 41, No. 8
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.8.3699-3705.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Rapid and Sensitive Assays for Determination of Hepatitis B Virus (HBV) Genotypes and Detection of HBV Precore and Core Promoter Variants

Munira Hussain,1 Chi-Jen Chu,1,{dagger} Erwin Sablon,2 and Anna S. F. Lok1*

Division of Gastroenterology, University of Michigan Medical Center, Ann Arbor, Michigan,1 Innogenetics NV, Ghent, Belgium2

Received 12 March 2003/ Returned for modification 5 May 2003/ Accepted 18 May 2003

Hepatitis B virus (HBV) genotypes may influence HBeAg seroconversion rates, mutational patterns in the precore (PC) and core promoter (CP) regions, severity of liver disease, and response to antiviral treatment. Development of rapid, simple, and standardized assays to detect viral genotypes and common mutations in the PC and CP regions can accelerate research on the clinical significance of these variants. We aim to assess the accuracy of a line probe assay in determining HBV genotypes and detecting HBV PC and CP variants. HBV genotypes in 701 patients and PC and CP variants in 600 patients with chronic HBV infection from China and the United States were studied using the INNO-LiPA assay. All but one (99.9%) sample were classified by the genotyping assay. All eight genotypes, i.e., A to H, were found. The INNO-LiPA genotyping assay results were completely concordant with those of sequencing. Using the INNO-LiPA PC assay, 99.8 and 94.7% samples were classifiable in the PC and CP regions, respectively. The PC assay results were completely concordant with those of sequencing in all samples that showed either wild-type or variant sequence. The line probe assay was more sensitive in detecting mixtures than was direct sequencing. By INNO-LiPA, only 50 and 27% of the samples, with mixed wild-type and variant sequence in the PC and CP region, respectively, showed mixed sequence by direct sequencing. INNO-LiPA is rapid, sensitive, and reliable—thus enabling accurate determination of HBV genotypes and detection of PC and CP variants in a large population of patients.

* 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.

{dagger} Present address: Taipei Veterans General Hospital, Taipei, Taiwan 11217, Republic of China.

Journal of Clinical Microbiology, August 2003, p. 3699-3705, Vol. 41, No. 8
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.8.3699-3705.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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