RecombinantHelicobacter bilisProtein P167 for Mouse Serodiagnosis in a Multiplex Microbead Assay
- 1 November 2004
- journal article
- research article
- Published by American Society for Microbiology in Clinical and Vaccine Immunology
- Vol. 11 (6) , 1094-1099
- https://doi.org/10.1128/cdli.11.6.1094-1099.2004
Abstract
Infection of mice withHelicobacter bilisis 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 ofH. bilisinfection. 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 fromH. bilisandHelicobacter hepaticus. For detectingH. bilisinfection 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 ofH. bilisinfection.Keywords
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