Scarman Anthony L, Chin James C, Earmens Graeme J, Delaney Stephen F, Djordjevic Steven P
Department of Biotechnology, University of New South Wales, sydney, NSW, Australia.
Microbiology and Immunology Section, Elizabeth Macarthur Agricultural Institute, Camden, NSW 2570, Australia.
Microbiology (Reading). 1997 Feb;143 ( Pt 2):663-673. doi: 10.1099/00221287-143-2-663.
Mycoplasma hyopneumoniae, M. hyorhinis and M. flocculare are commonly isolated from the respiratory tract of pigs and are phylogenetically related. The identification and characterization of antigens specific for M. hyopneumoniae 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 M. hyopneumoniae, four strains of M. hyorhinis and a type strain of M. flocculare were compared using SDS-PAGE and immunoblotting. Five strains of M. hyopneumoniae 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 M. hyopneumoniae may possess a structurally modified repertoire of antigens. Major M. hyopneumoniae 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 M. hyopneumoniae. Porcine hyperimmune sera raised against the GDL type strain of M. hyorhinis reacted strongly with all M. hyorhinis 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 immunoreactivity 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 M. hyopneumoniae-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-42 kDa serum reacted specifically with a 43 kDa antigen in whole cell lysates of geographically diverse strains of M. hyopneumoniae and failed to cross-react with M. flocculare or M. hyorhinis 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.
猪肺炎支原体、猪鼻支原体和絮状支原体通常从猪的呼吸道中分离出来，并且在系统发育上相关。鉴定和表征猪肺炎支原体特异性抗原对于血清学试剂的开发以及理解该病原体的致病机制至关重要。使用SDS-PAGE和免疫印迹比较了6株猪肺炎支原体、4株猪鼻支原体和1株絮状支原体的蛋白质和抗原谱。最初从不同地理区域分离的5株猪肺炎支原体产生了相似的蛋白质和抗原谱。其中一株C1735/2产生了独特的蛋白质谱且免疫反应性较差，这表明一些猪肺炎支原体菌株可能具有结构修饰的抗原库。使用针对猪肺炎支原体高代和低代菌株产生的超免疫猪血清进行免疫印迹，鉴定出了分子量约为36、43、48、52、76、78、80、82、94、106、114和200 kDa的主要猪肺炎支原体抗原。针对猪鼻支原体GDL型菌株产生的猪超免疫血清与所有猪鼻支原体菌株都有强烈反应，尽管其图谱显示出相当大的差异。在GDL和BTS-7型菌株以及田间菌株2中鉴定出了分子量为42、49、52、78、80和82 kDa的主要抗原；然而，田间菌株1产生了独特的图谱。开发了一种制备性SDS-PAGE分析（PPP）技术，该技术能够通过ELISA定量变性抗原与猪血清的免疫反应性。PPP有助于快速鉴定这三种支原体物种之间的种特异性和交叉反应性抗原。PPP研究揭示了几种强免疫反应性的猪肺炎支原体特异性抗原，分子量分别为43、76、94、114和200 kDa，以及分子量在52至62 kDa之间的抗原，这些抗原在免疫印迹研究中并不明显。兔抗42 kDa单特异性血清与来自不同地理区域的猪肺炎支原体菌株的全细胞裂解物中的43 kDa抗原特异性反应，并且不与絮状支原体或猪鼻支原体全细胞裂解物发生交叉反应。本研究鉴定出了许多猪肺炎支原体特异性抗原，这些抗原值得进一步研究，以确定它们作为诊断试剂的潜力以及它们在致病性中所起的作用（如果有的话）。
