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Clinical and Diagnostic Laboratory Immunology, November 2004, p. 1094-1099, Vol. 11, No. 6
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.6.1094-1099.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Recombinant Helicobacter bilis Protein P167 for Mouse Serodiagnosis in a Multiplex Microbead Assay

Sunlian Feng,1 Lon V. Kendall,2 Emir Hodzic,1 Scott Wong,1 Edward Lorenzana,1 Kimberly Freet,1 Karin S. Ku,1 Paul A. Luciw,1,3 Stephen W. Barthold,1 and Imran H. Khan1*

Center for Comparative Medicine,1 Comparative Pathology Laboratory,2 Department of Medical Pathology and Laboratory Medicine, University of California, Davis, California3

Received 25 May 2004/ Returned for modification 8 July 2004/ Accepted 30 July 2004

Infection of mice with Helicobacter bilis is widespread in research and commercial mouse colonies. Therefore, sensitive, specific, and high-throughput assays are needed for rapid and accurate testing of mice in large numbers. This report describes a novel multiplex assay, based on fluorescent microbeads, for serodetection of H. bilis infection. The assay requires only a few microliters of serum to perform and is amenable to a high-throughput format. Individual microbead sets were conjugated to purified, H. bilis-specific, recombinant proteins P167C and P167D and bacterial membrane extracts from H. bilis and Helicobacter hepaticus. For detecting H. bilis infection in the microbead multiplex assay, P167C and P167D provided significantly higher sensitivities (94 and 100%, respectively) and specificities (100 and 95%, respectively) than membrane extract (78% sensitivity and 65% specificity). Microbead multiplex assay results were validated by enzyme-linked immunosorbent assay. Purified recombinant proteins showed low batch-to-batch variation; this feature allows for ease of quality control, assay robustness, and affordability. Thus, recombinant antigens are highly suitable in the multiplex microbead assay format for serodetection of H. bilis infection.

* Corresponding author. Mailing address: Center for Comparative Medicine, University of California at Davis, One Shields Ave., Davis, CA 95616. Phone: (530) 752-1245. Fax: (530) 752-7914. E-mail: ihkhan{at}ucdavis.edu.

Clinical and Diagnostic Laboratory Immunology, November 2004, p. 1094-1099, Vol. 11, No. 6
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.6.1094-1099.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Khan, I. H., Mendoza, S., Yee, J., Deane, M., Venkateswaran, K., Zhou, S. S., Barry, P. A., Lerche, N. W., Luciw, P. A. (2006). Simultaneous Detection of Antibodies to Six Nonhuman-Primate Viruses by Multiplex Microbead Immunoassay. CVI 13: 45-52 [Abstract] [Full Text]  


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