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使用ESAT-6/MPB70/MPB83重组嵌合体的酶联免疫吸附测定法用于自然感染牛中牛分枝杆菌的诊断。

ELISA using a recombinant chimera of ESAT-6/MPB70/MPB83 for Mycobacterium bovis diagnosis in naturally infected cattle.

作者信息

Souza Ingrid Ieda Fernando de, Rodrigues Rudielle de Arruda, Gonçalves Jorge Kláudia Santos, Silva Márcio Roberto, Lilenbaum Walter, Vidal Carlos Eugênio Soto, Etges Rodrigo Nestor, Kostovic Miladin, Araújo Flábio Ribeiro

机构信息

Postgraduate Program in Biotechnology and Biodiversity of the Central Western Region, Federal University of Mato Grosso do Sul, Campo Grande, MS 79070-900, Brazil.

Postgraduate Program in Veterinary Sciences, School of Veterinary Medicine (FAMEZ), Federal University of Mato Grosso do Sul, Campo Grande, MS 79070-900, Brazil.

出版信息

J Vet Med Sci. 2019 Jan 8;81(1):9-14. doi: 10.1292/jvms.18-0364. Epub 2018 Oct 11.

Abstract

Bovine tuberculosis (bTB) control programs generally rely on intradermal tuberculin tests for the antemortem diagnosis of Mycobacterium bovis infection in cattle, but these tests detect only a portion of the infected animals. The aim of the present study was to evaluate the diagnostic coverage of a combination of the bTB antemortem techniques known as the comparative intradermal tuberculin test (CITT) and an ELISA based on a recombinant chimera of ESAT-6/MPB70/MPB83 as the antigen in cattle. The results were compared to postmortem findings based on M. bovis culturing and PCR. Paired comparisons of all data (n=92) demonstrated that ELISA and LST results compared to the culturing results did not present significant differences (P=0.27 on McNemar's test and P=0.12 on Fisher's exact test, respectively). Using culturing as the gold standard, the sensitivity and specificity of ELISA were 79.5% (95% CI: 64.5-89.2%) and 75.5% (95% CI: 62.4-85.1%), respectively, whereas LST demonstrated 100% sensitivity (95% CI: 91.03-100%) and 92.5% specificity (95% CI: 82.1-97.0%). The ELISA results did not reveal significant differences in relation to the LST results (P>0.99 on Fisher's exact test). Using the latter as the gold standard, the sensitivity and specificity of ELISA were 79.1% (95% CI: 64.8-88.6%) and 79.6% (95% CI: 66.4-88.5%), respectively. The use of ELISA with the recombinant chimera of ESAT-6/MPB70/MPB83 as the antigen complements the diagnostic coverage provided by CITT and increases the removal of infected animals from herds.

摘要

牛结核病(bTB)防控计划通常依靠皮内结核菌素试验对牛群中牛分枝杆菌感染进行生前诊断,但这些试验只能检测出一部分受感染动物。本研究的目的是评估一种牛结核病生前诊断技术组合的诊断覆盖率,该组合包括比较皮内结核菌素试验(CITT)和以ESAT-6/MPB70/MPB83重组嵌合体为抗原的酶联免疫吸附测定(ELISA)。将结果与基于牛分枝杆菌培养和聚合酶链反应(PCR)的死后检测结果进行比较。对所有数据(n = 92)进行配对比较表明,ELISA和皮内结核菌素试验结果与培养结果相比,无显著差异(分别在McNemar检验中P = 0.27,在Fisher精确检验中P = 0.12)。以培养作为金标准,ELISA的敏感性和特异性分别为79.5%(95%置信区间:64.5 - 89.2%)和75.5%(95%置信区间:62.4 - 85.1%),而皮内结核菌素试验显示敏感性为100%(95%置信区间:91.03 - 100%),特异性为92.5%(95%置信区间:82.1 - 97.0%)。ELISA结果与皮内结核菌素试验结果无显著差异(在Fisher精确检验中P>0.99)。以后者作为金标准,ELISA的敏感性和特异性分别为79.1%(95%置信区间:64.8 - 88.6%)和79.6%(95%置信区间:66.4 - 88.5%)。使用以ESAT-6/MPB70/MPB83重组嵌合体为抗原的ELISA可补充CITT提供的诊断覆盖率,并增加从牛群中清除受感染动物的数量。

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