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通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)和免疫印迹法检测猪肺炎支原体菌株间的蛋白质和抗原变异性。

Protein and antigenic variability among Mycoplasma hyopneumoniae strains by SDS-PAGE and immunoblot.

作者信息

Assunção P, De la Fe C, Ramírez A S, González Llamazares O, Poveda J B

机构信息

Unidad de Epidemiologia y Medicina Preventiva, Facultad de Veterinaria, Universidad de Las Palmas de Gran Canaria, Trasmontaña s/n., 35416, Arucas, Spain.

出版信息

Vet Res Commun. 2005 Oct;29(7):563-74. doi: 10.1007/s11259-005-3083-5.

Abstract

Porcine enzootic pneumonia (PEP), with Mycoplasma hyopneumoniae as the primary agent, is a chronic respiratory disease that causes major economic losses to the pig industry worldwide. The aim of this work was to analyse 18 field strains of M. hyopneumoniae isolated in Gran Canaria (Spain) and the reference M. hyopneumoniae strain by SDS-PAGE and immunoblot. A monoclonal antibody (MAb) against the membrane protein p46 reacted with all the strains in this study. In contrast, a purified polyclonal antibody (PAb) against the cytoplasmic protein p36 reacted with this protein in only 10 strains. A MAb against the adhesin protein p97 stained multiple proteins of different sizes and with different intensities. Different antigenic patterns in the same M. hyopneumoniae strains were also observed after different numbers of passages in culture medium. Furthermore, variability in the staining of the 36 kDa protein was observed, depending on whether the p36 PAb or the antiserum against M. hyopneumoniae reference strain was used. It is concluded that local M. hyopneumoniae field isolates in Gran Canaria are characterized by protein diversity.

摘要

猪地方性肺炎(PEP),主要由猪肺炎支原体引起,是一种慢性呼吸道疾病,给全球养猪业造成重大经济损失。本研究的目的是通过SDS - PAGE和免疫印迹法分析在大加那利岛(西班牙)分离的18株猪肺炎支原体田间菌株以及猪肺炎支原体参考菌株。一种针对膜蛋白p46的单克隆抗体(MAb)与本研究中的所有菌株发生反应。相比之下,一种针对细胞质蛋白p36的纯化多克隆抗体(PAb)仅与10株菌株中的该蛋白发生反应。一种针对黏附蛋白p97的单克隆抗体对不同大小和强度的多种蛋白进行了染色。在培养基中传代不同次数后,同一猪肺炎支原体菌株中也观察到了不同的抗原模式。此外,根据使用的是p36 PAb还是针对猪肺炎支原体参考菌株的抗血清,观察到36 kDa蛋白染色的变异性。得出的结论是,大加那利岛当地的猪肺炎支原体田间分离株具有蛋白质多样性的特征。

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