Recombinant Helicobacter bilis protein P167 for mouse serodiagnosis in a multiplex microbead assay

Sunlian Feng, Lon V. Kendall, Emir Hodzic, Scott Wong, Edward Lorenzana, Kimberly Freet, Karin S. Ku, Paul A Luciw, Stephen W Barthold, Imran Khan

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)1094-1099
Number of pages6
JournalClinical and Diagnostic Laboratory Immunology
Volume11
Issue number6
DOIs
StatePublished - Nov 2004

ASJC Scopus subject areas

  • Microbiology (medical)
  • Immunology and Allergy
  • Clinical Biochemistry
  • Immunology

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