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Clinical and Vaccine Immunology, December 2007, p. 1616-1622, Vol. 14, No. 12
1071-412X/07/$08.00+0     doi:10.1128/CVI.00345-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Use of Recombinant Antigens To Detect Antibodies against Mycoplasma suis, with Correlation of Serological Results to Hematological Findings{triangledown}

Katharina Hoelzle,1 Julia Grimm,2 Mathias Ritzmann,2 Karl Heinritzi,2 Paul Torgerson,3 Anja Hamburger,1 Max M. Wittenbrink,1 and Ludwig E. Hoelzle1*

Institute of Veterinary Bacteriology, Vetsuisse Faculty, University of Zurich, Zurich, Switzerland,1 Clinic for Swine, Ludwig-Maximilians-University Munich, Munich, Germany,2 Institute of Parasitology, Vetsuisse Faculty, University of Zurich, Zurich, Switzerland3

Received 19 August 2007/ Returned for modification 7 September 2007/ Accepted 7 October 2007

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.

* Corresponding author. Mailing address: Institute of Veterinary Bacteriology, Vetsuisse Faculty, University of Zurich, Winterthurerstrasse 270, 8057 Zurich, Switzerland. Phone: 41-44 635 8613. Fax: 41-44 635 8912. E-mail: lhoelzle{at}vetbakt.uzh.ch

{triangledown} Published ahead of print on 17 October 2007.

Clinical and Vaccine Immunology, December 2007, p. 1616-1622, Vol. 14, No. 12
1071-412X/07/$08.00+0     doi:10.1128/CVI.00345-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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