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用于诊断绵羊流产嗜衣原体感染的八种血清学检测方法的比较评估

Comparative evaluation of eight serological assays for diagnosing Chlamydophila abortus infection in sheep.

作者信息

Wilson Kim, Livingstone Morag, Longbottom David

机构信息

Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik EH26 OPZ, UK.

出版信息

Vet Microbiol. 2009 Mar 16;135(1-2):38-45. doi: 10.1016/j.vetmic.2008.09.043. Epub 2008 Sep 16.

Abstract

Chlamydophila abortus is one of the principal causes of late-term abortion (enzootic abortion of ewes or EAE) in sheep across Europe. Serological diagnosis of EAE is routinely carried out by the complement fixation test, although the interpretation of results can often be difficult because of cross reaction with Chlamydophila pecorum, which also commonly infects sheep. The purpose of this study was to evaluate and compare four ELISAs developed at Moredun Research Institute and based on whole C. abortus elementary bodies (EBs), an outer membrane preparation of the whole organism (SolPr) and two recombinant polymorphic outer membrane protein fragments (rOMP90-3 and rOMP90-4), with 3 commercial tests, the CHEKIT Chlamydophila Abortus, Pourquier ELISA Chlamydophila abortus and ImmunoComb Ovine Chlamydophila Antibody tests. The tests were evaluated using a panel of 202 sera from experimentally and naturally infected animals, as well as from EAE-free flocks. The EB, SolPr and CHEKIT ELISAs performed similarly to the CFT, all lacking in specificity by cross reacting with sera from C. pecorum infected animals. The ImmunoComb also lacked specificity with C. pecorum sera, but also badly cross reacted with sera from EAE-free flocks. The rOMP90-3, rOMP90-4 and Pourquier ELISAs were the most specific, although the Pourquier test appeared less sensitive with sera from naturally infected animals. Overall, the rOMP90-3 ELISA performed the best, with high sensitivity (96.8%) and no cross reaction with sera from C. pecorum infected animals or from EAE-free flocks (100% specificity) and so would be a suitable alternative to the CFT for the serological diagnosis of EAE.

摘要

流产嗜衣原体是欧洲绵羊晚期流产(母羊地方流行性流产或EAE)的主要病因之一。EAE的血清学诊断通常通过补体结合试验进行,不过由于与同样常感染绵羊的嗜肺衣原体发生交叉反应,结果的解读往往很困难。本研究的目的是评估和比较莫雷登研究所开发的四种基于流产嗜衣原体全原生小体(EBs)、全菌体的外膜制剂(SolPr)以及两种重组多态性外膜蛋白片段(rOMP90-3和rOMP90-4)的酶联免疫吸附测定(ELISA),与三种商业检测方法,即CHEKIT流产嗜衣原体检测、普尔基埃尔流产嗜衣原体ELISA和免疫组合绵羊流产嗜衣原体抗体检测。使用一组来自实验感染和自然感染动物以及未发生EAE的羊群的202份血清对这些检测方法进行评估。EB、SolPr和CHEKIT ELISA的表现与补体结合试验相似,都因与嗜肺衣原体感染动物的血清发生交叉反应而缺乏特异性。免疫组合检测对嗜肺衣原体血清也缺乏特异性,而且还与未发生EAE的羊群的血清发生严重交叉反应。rOMP90-3、rOMP90-4和普尔基埃尔ELISA的特异性最高,不过普尔基埃尔检测对自然感染动物的血清似乎不太敏感。总体而言,rOMP90-3 ELISA表现最佳,具有高灵敏度(96.8%),且与嗜肺衣原体感染动物或未发生EAE的羊群的血清无交叉反应(特异性为100%),因此可作为补体结合试验用于EAE血清学诊断的合适替代方法。

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