title: Diagnosis of babesiosis using an immunoblot serologic test.
abstract: Although the current indirect immunofluorescent assay (IFA) diagnostic antibody test for human babesiosis is sensitive and specific, an immunoblot antibody test may be easier to standardize and to perform. Our objective, therefore, was to determine the efficacy of and develop interpretive criteria for an immunoblot antibody test for diagnosing acute human babesiosis using a Babesia microti whole-cell lysate as the antigen. We compared the reactivity of sera to a B. microti immunoblot assay in 24 human subjects experiencing symptoms and expressing laboratory evidence of babesiosis, 28 subjects who experienced Lyme disease, 12 subjects who experienced human granulocytic ehrlichiosis, and 51 subjects who reported no history of any of these diseases and whose sera did not react against B. microti antigen in an IFA test. Immunoblot strips were impregnated with proteins derived from the GI strain of B. microti that had been electrophoresed in an acrylamide sodium dodecyl sulfate gel, followed by electroblotting onto nitrocellulose membranes. The sera of all subjects who experienced babesiosis reacted against the B. microti antigen in the IFA and against at least one of nine immunoblot protein bands specific to B. microti. In contrast, none of the sera from people who appeared not to have experienced this infection reacted against the B. microti antigen in the IFA (compared to 4% in the immunoblot assay). When two reactive bands were considered as definitive, immunoblot test sensitivity was 96%, while specificity was 99% and predictive positivity and predictive negativity were 96 and 99%, respectively. Our B. microti immunoblot procedure shows promise as a sensitive, specific, and reproducible assay for routine clinical diagnosis of acute babesiosis.
authors.affiliation: Department of Clinical Microbiology, University of Connecticut School of Medicine, Farmington, Connecticut 06030, USA, Rryan@NSO1.UCHC.edu
authors.forename: R
authors.initials: R
authors.lastname: Ryan
authors.forename: P J
authors.initials: PJ
authors.lastname: Krause
authors.forename: J
authors.initials: J
authors.lastname: Radolf
authors.forename: K
authors.initials: K
authors.lastname: Freeman
authors.forename: A
authors.initials: A
authors.lastname: Spielman
authors.forename: R
authors.initials: R
authors.lastname: Lenz
authors.forename: A
authors.initials: A
authors.lastname: Levin
mesh_terms: D000913:Antibodies, Protozoan / Q000032:analysis / Q000097:blood; D000953:Antigens, Protozoan / Q000276:immunology*; D001404:Babesiosis / Q000175:diagnosis* / Q000276:immunology; D016873:Ehrlichiosis / Q000276:immunology; D000939:Epitopes; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D015151:Immunoblotting / Q000379:methods* / Q000592:standards; D008193:Lyme Disease / Q000276:immunology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D000939:Epitopes