Multiplex Screening of Surface Proteins from Mycoplasma mycoides subsp mycoides Small Colony for an Antigen Cocktail Enzyme-Linked Immunosorbent Assay
2009 (English)In: Clinical and Laboratory Immunology, ISSN 1556-6811, E-ISSN 1556-679X, Vol. 16, no 11, 1665-1674 p.Article in journal (Refereed) Published
A recombinant antigen cocktail ELISA for diagnosis of contagious bovine pleuropneumonia (CBPP) was developed after careful selection of antigens among one third of the surface proteome of the infectious agent Mycoplasma mycoides subsp. mycoides SC (M. mycoides SC). First, a miniaturized and parallelized assay system employing antigen suspension bead array technology was used to screen 97 bovine sera for their humoral immune response towards 61 recombinant surface proteins from M. mycoides SC. Statistical analysis of the data resulted in selection of eight proteins that showed strong serologic responses in CBPP-affected sera and minimal reactivity in negative control sera, with p-values less than 10-6. Only minor cross reactivity to hyperimmune sera against other mycoplasmas was observed. When applied in an ELISA, the cocktail of eight recombinant antigens allowed a five fold signal separation between 24 CBPP-affected and 23 CBPP-free sera from different geographical origin. No false positives and only two false negatives were obtained. In conclusion, the selected recombinant mycoplasma antigens qualified as highly specific markers for CBPP and could be employed in both a suspension bead array platform and a cocktail ELISA setting. This set of proteins and technologies therefore offer a powerful combination to drive and further improve serological assays towards reliable, simple and cost-effective diagnosis of CBPP.
Place, publisher, year, edition, pages
Washington D.C.: American Society for Microbiology , 2009. Vol. 16, no 11, 1665-1674 p.
contagious bovine pleuropneumonia, lipoprotein lppq, genetic-characterization, sc, diagnosis, elisa, cbpp, capricolum, tanzania, cluster
IdentifiersURN: urn:nbn:se:kth:diva-11105DOI: 10.1128/CVI.00223-09ISI: 000271373700019ScopusID: 2-s2.0-70350702934OAI: oai:DiVA.org:kth-11105DiVA: diva2:235811
QC 20100719. Updated from in press to published. Previous title "Multiplex screening of surface proteins from Mycoplasma mycoides
SC for an antigen cocktail ELISA"2009-09-182009-09-182013-02-08Bibliographically approved