Characterization of Mycoplasma mycoides subsp. capri isolates by SDS-PAGE, immunoblotting and PFGE

The purpose of this study was to analyze a bank of 35 Mycoplasma mycoides subsp. capri (Mmc) isolates collected from contagious agalactia outbreaks during a 9-year period (2004-2012) in Sardinia, Italy. Isolates were characterized using genotyping and phenotyping methods. For each isolate, pulsed field gel electrophoresis (PFGE) DNA profile was determined. The PFGE study, performed with the rare-cutting restriction enzyme BnlI, showed a high degree of genetic heterogeneity. Our findings show the value of PFGE analysis for revealing the genetic diversity of isolates and its usefulness for epidemiology investigations. In addition, the whole cell protein profiles of each isolate were studied by Coomassie Blue-stained sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot (IB). After IB with pooled serum from naturally infected goats, two immunogenic proteins of about 90 and 60 kDa were identified in all isolates analyzed. These proteins may have potential for the development of serological diagnostic tests.