del Campo Sepúlveda E M, Altman E, Kobisch M, D'Allaire S, Gottschalk M
Groupe de Recherche sur les Maladies Infectieuses du Porc, Faculté de Médecine Vétérinaire, Université de Montréal, Québec, Canada.
Vet Microbiol. 1996 Sep;52(1-2):113-25. doi: 10.1016/0378-1135(96)00056-9.
In the present study a purified capsular polysaccharide antigen-based indirect ELISA (CPS-ELISA) to detect antibodies against Streptococcus suis capsular type 2 was developed and compared with a whole cell antigen-based ELISA (WCA-ELISA). The WCA-ELISA presented a very low specificity when rabbit antisera to other capsular types were tested. Most of these cross-reactions were due to common proteins. The standardized CPS-ELISA gave satisfactory results using a concentration of 0.1 micrograms/well; most cross-reactions decreased significantly, with some exceptions, such as those shared by capsular types 1/2, 12 and 17. These cross-reactions were mainly due to common epitopes present in the capsule, as shown by immunoblotting. In a second experiment, the CPS-ELISA was used to detect antibodies in experimentally infected piglets. Despite the fact that capsular type 2 S. suis could be reisolated from all infected animals during and/or after the trial, antibody titers against a second infection. Sera from piglets experimentally infected were completely protected against a second infection. Sera from piglets experimentally infected with S. suis capsular types 1/2 or 12 presented cross-reactions at low dilutions, confirming data previously obtained with rabbit sera. Finally, sera of animals from herds with clinical signs associated with S. suis capsular type 2 did not present titers significantly different from those of disease free herds. From our results we concluded that the CPS-ELISA developed in this study can not be used as a diagnostic tool to identify infected animals.
在本研究中,开发了一种基于纯化荚膜多糖抗原的间接ELISA(CPS-ELISA)来检测抗猪链球菌2型荚膜的抗体,并与基于全细胞抗原的ELISA(WCA-ELISA)进行比较。当检测针对其他荚膜型的兔抗血清时,WCA-ELISA的特异性非常低。这些交叉反应大多是由共同蛋白引起的。标准化的CPS-ELISA在使用浓度为0.1微克/孔时给出了满意的结果;大多数交叉反应显著减少,但有一些例外,如荚膜型1/2、12和17之间共有的交叉反应。免疫印迹显示,这些交叉反应主要是由于荚膜中存在的共同表位。在第二个实验中,CPS-ELISA用于检测实验感染仔猪体内的抗体。尽管在试验期间和/或试验后可以从所有感染动物中重新分离出2型猪链球菌荚膜,但针对第二次感染的抗体滴度。实验感染的仔猪血清对第二次感染具有完全保护作用。实验感染1/2型或12型猪链球菌荚膜的仔猪血清在低稀释度下出现交叉反应,证实了先前用兔血清获得的数据。最后,来自与2型猪链球菌荚膜相关临床症状的猪群的动物血清滴度与无病猪群的血清滴度没有显著差异。根据我们的结果,我们得出结论,本研究中开发的CPS-ELISA不能用作识别感染动物的诊断工具。