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使用单克隆抗体鉴定胸膜肺炎放线杆菌7型和4型菌株:证明与里氏放线杆菌存在共同的脂多糖O链表位

Identification of Actinobacillus pleuropneumoniae strains of serotypes 7 and 4 using monoclonal antibodies: demonstration of common LPS O-chain epitopes with Actinobacillus lignieresii.

作者信息

Lebrun A, Lacouture S, Côté D, Mittal K R, Gottschalk M

机构信息

Groupe de Recherche sur les Maladies Infectieuses du Porc, Faculté de médecine vétérinaire, Université de Montréal, St. Hyacinthe, Qué, Canada.

出版信息

Vet Microbiol. 1999 Mar 19;65(4):271-82. doi: 10.1016/s0378-1135(98)00301-0.

Abstract

Monoclonal antibodies (Mabs) against Actinobacillus pleuropneumoniae serotype 7 were produced and characterized. Three Mabs directed against surface polysaccharides were selected. One of the Mabs was directed against a capsular polysaccharide epitope (CPS) of A. pleuropneumoniae serotype 7 whereas two other Mabs reacted with different epitopes of the LPS O-chain. One of the latter reacted with the reference strain of serotype 7 and the other one with serotypes 7 and 4. These three Mabs were used to test, by Dot-ELISA, 508 field strains of A. pleuropneumoniae. None of the strains belonging to other serotypes different from serotypes 4 and 7 were positive with the Mabs. Used in combination, the CPS and one of the LPS O-chain directed Mabs were shown to be suitable for serotyping since they detected 100% of serotype 7 strains. In this study, we confirm for the first time that A. pleuropneumoniae serotype 4 is present in North America. Finally, both O-chain specific Mabs also reacted with the O-chain of Actinobacillus lignieresii. The cross-reactivity between the two species was confirmed using sera from pigs experimentally infected with A. pleuropneumoniae serotype 7 and A. lignieresii, using immunoblotting and ELISA. This is the first report of a specific cross-reactivity between the LPS of these bacterial species.

摘要

制备并鉴定了抗胸膜肺炎放线杆菌7型的单克隆抗体(Mabs)。选择了三种针对表面多糖的单克隆抗体。其中一种单克隆抗体针对胸膜肺炎放线杆菌7型的荚膜多糖表位(CPS),而另外两种单克隆抗体与脂多糖O链的不同表位发生反应。后一种中的一种与7型参考菌株反应,另一种与7型和4型反应。这三种单克隆抗体用于通过斑点酶联免疫吸附测定(Dot-ELISA)检测508株胸膜肺炎放线杆菌的田间菌株。与4型和7型不同的其他血清型的菌株用这些单克隆抗体检测均为阴性。联合使用时,CPS和一种针对脂多糖O链的单克隆抗体被证明适用于血清分型,因为它们检测到了100%的7型菌株。在本研究中,我们首次证实北美存在胸膜肺炎放线杆菌4型。最后,两种O链特异性单克隆抗体也与里氏放线杆菌的O链发生反应。使用来自经胸膜肺炎放线杆菌7型和里氏放线杆菌实验感染的猪的血清,通过免疫印迹和酶联免疫吸附测定证实了这两个菌种之间的交叉反应性。这是这些细菌菌种脂多糖之间特异性交叉反应性的首次报道。

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