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Volume 143, Issue 2

Identification of novel species-specific antigens of by preparative SDS-PAGE ELISA profiling Free

Abstract

and are commonly isolated from the respiratory tract of pigs and are phylogenetically related. The identification and characterization of antigens specific for is crucial for the development of serological reagents and for understanding the mechanisms of pathogenicity of this pathogen. Protein and antigen profiles of six strains of four strains of and a type strain of were compared using SDS-PAGE and immunoblotting. Five strains of originally isolated from diverse geographical regions produced similar protein and antigen profiles. One strain, C1735/2, produced a unique protein profile and was poorly immunoreactive, suggesting that some strains of may possess a structurally modified repertoire of antigens. Major antigens with molecular masses of approximately 36, 43, 48, 52, 76, 78, 80, 82, 94, 106, 114 and 200 kDa were identified by immunoblotting using hyperimmune pig sera raised against both high and low passage strains of Porcine hyperimmune sera raised against the GDL type strain of reacted strongly with all strains although the profiles displayed considerable variation. Major antigens of molecular mass 42, 49, 52, 78, 80 and 82 kDa were identified in type strains GDL and BTS-7 and field strain 2; however, field strain 1 produced a unique profile. A preparative SDS-PAGE profiling (PPP) technique was developed which enabled quantification of the immiunoreactivity of denatured antigens with porcine serum by ELISA. PPP facilitated the rapid identification of species-specific and cross-reactive antigens among the three mycoplasma species. PPP studies revealed several strongly immunoreactive -specific antigens of 43, 76, 94, 114 and 200 kDa as well as antigens of molecular mass between 52 and 62 kDa which were not apparent in immunoblotting studies. Rabbit monospecific anti-43 kDa serum reacted specifically with a 43 kDa antigen in whole cell lysates of geographically diverse strains of and failed to cross-react with or whole cell lysates. This study has identified a number of M. hyopneumoniae-specific antigens which warrant further investigation to determine their potential as diagnostic reagents and the role they play, if any, in pathogenicity.

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Keyword(s): diagnostic antigens , immunoblotting , Mycoplasma hyopneumoniae , Mycoplasma hyorhinis and preparative SDS-PAGE profiling
© Society for General Microbiology 1997
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1997-02-01
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Identification of novel species-specific antigens of Mycoplasma hyopneumoniae by preparative SDS-PAGE ELISA profiling
Microbiology 143, 663 (1997); https://doi.org/10.1099/00221287-143-2-663
/content/journal/micro/10.1099/00221287-143-2-663
/content/journal/micro/10.1099/00221287-143-2-663
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