title: Use of recombinant antigens to detect antibodies against Mycoplasma suis, with correlation of serological results to hematological findings.
abstract: Porcine eperythrozoonosis is a disease with worldwide distribution caused by the unculturable hemotrophic bacterium Mycoplasma suis. Current serological testing utilizes crude M. suis antigens purified from the blood of experimentally infected pigs. These antigens show high variability and are restricted to specialized laboratories. We evaluated a novel serological assay based on two recombinant M. suis antigens (rMSG1 and rHspA1). Antigen specificity was proven by means of sera raised against nonhemotrophic mycoplasma and other relevant bacteria. Using experimental and convalescent-phase sera, rMSG1 and rHspA1 enzyme-linked immunosorbent assays (ELISAs) demonstrated sensitivities, specificities, and predictive values (94.0 to 100.0%) equal to or higher than those of the M. suis whole-cell ELISA. Field samples from 120 weaning piglets grouped by quantitative PCR results were used to evaluate the diagnostic capability of the new ELISA systems in comparison to that of the whole-cell ELISA. Assuming a 100.0% specificity of the PCR, the whole-cell ELISA, rHspA1 ELISA, and rMSG1 ELISA showed specificities of 84.8%, 83.8%, and 90.6% and sensitivities of 61.5%, 74.0% and 58.1%, respectively. Cohen's kappa coefficients comparing the recombinant ELISAs to the whole-cell ELISA indicate moderate to substantial agreement. The detection of anti-MSG1 and/or anti-HspA1 antibodies in pigs was significantly correlated with decreased hematocrit, erythrocyte numbers, and hemoglobin concentrations, indicating that a single seropositive result is connected with clinical and etiological significance. In conclusion, rMSG1 and rHspA1 are sensitive and specific serological and infection markers which are for the first time used independently of animal experiments. They are especially fit to be used in routine diagnosis, pathogenesis studies, and large-scale epidemiological investigations.
authors.affiliation: Institute of Veterinary Bacteriology, Vetsuisse Faculty, University of Zurich, Winterthurerstrasse 270, 8057 Zurich, Switzerland. lhoelzle@vetbakt.uzh.ch
authors.forename: Katharina
authors.initials: K
authors.lastname: Hoelzle
authors.forename: Julia
authors.initials: J
authors.lastname: Grimm
authors.forename: Mathias
authors.initials: M
authors.lastname: Ritzmann
authors.forename: Karl
authors.initials: K
authors.lastname: Heinritzi
authors.forename: Paul
authors.initials: P
authors.lastname: Torgerson
authors.forename: Anja
authors.initials: A
authors.lastname: Hamburger
authors.forename: Max M
authors.initials: MM
authors.lastname: Wittenbrink
authors.forename: Ludwig E
authors.initials: LE
authors.lastname: Hoelzle
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology*; D000917:Antibody Formation / Q000276:immunology; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D015415:Biomarkers / Q000097:blood; D004247:DNA / Q000097:blood; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D004909:Erythrocyte Indices / Q000662:veterinary*; D004926:Escherichia coli / Q000235:genetics; D015151:Immunoblotting / Q000662:veterinary; D007700:Kinetics; D009174:Mycoplasma / Q000145:classification / Q000276:immunology* / Q000472:pathogenicity; D009175:Mycoplasma Infections / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011237:Predictive Value of Tests; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000662:veterinary; D013045:Species Specificity; D013156:Splenectomy; D013223:Statistics as Topic; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D015415:Biomarkers; D011993:Recombinant Fusion Proteins; D004247:DNA