title: Recombinant Helicobacter bilis protein P167 for mouse serodiagnosis in a multiplex microbead assay.
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.
authors.affiliation: Center for Comparative Medicine, University of California at Davis, One Shields Ave., Davis, CA 95616, USA.
authors.forename: Sunlian
authors.initials: S
authors.lastname: Feng
authors.forename: Lon V
authors.initials: LV
authors.lastname: Kendall
authors.forename: Emir
authors.initials: E
authors.lastname: Hodzic
authors.forename: Scott
authors.initials: S
authors.lastname: Wong
authors.forename: Edward
authors.initials: E
authors.lastname: Lorenzana
authors.forename: Kimberly
authors.initials: K
authors.lastname: Freet
authors.forename: Karin S
authors.initials: KS
authors.lastname: Ku
authors.forename: Paul A
authors.initials: PA
authors.lastname: Luciw
authors.forename: Stephen W
authors.initials: SW
authors.lastname: Barthold
authors.forename: Imran H
authors.initials: IH
authors.lastname: Khan
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D000942:Antigens, Bacterial / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D019084:Fluorescent Antibody Technique, Indirect / Q000379:methods; D016998:Helicobacter / Q000235:genetics / Q000276:immunology*; D016481:Helicobacter Infections / Q000175:diagnosis* / Q000662:veterinary*; D051379:Mice; D011237:Predictive Value of Tests; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012376:Rodent Diseases / Q000175:diagnosis* / Q000382:microbiology
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D011994:Recombinant Proteins