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比较分析不同血清学和分子检测方法在检测比利时绵羊和山羊小反刍兽疫病毒(SRLV)中的应用。

Comparative Analysis of Different Serological and Molecular Tests for the Detection of Small Ruminant Lentiviruses (SRLVs) in Belgian Sheep and Goats.

机构信息

Unit of Enzootic, Vector-Borne and Bee Diseases, Sciensano, Groeselenberg 99, 1180 Brussels, Belgium.

Dierengezondheidszorg Vlaanderen (DGZ), Industrielaan 29, 8820 Torhout, Belgium.

出版信息

Viruses. 2018 Dec 8;10(12):696. doi: 10.3390/v10120696.

Abstract

Countries rely on good diagnostic tests and appropriate testing schemes to fight against economically important small ruminant lentivirus (SRLV) infections. We undertook an extensive comparative analysis of seven commercially available serological tests and one in-house real-time PCR (qPCR) detecting genotype A and B strains using a large panel of representative Belgian field samples and samples from experimentally infected sheep and goats. ELISAs generally performed well and detected seroconversion within three weeks post experimental infection. Two enzyme-linked immunosorbent assays (ELISAs) (Elitest and IDscreen kits) showed the highest sensitivities (>96%) and specificities (>95%) in both species, and their combined use allowed to correctly identify the infection status of all animals. Individual agar gel immunodiffusion (AGIDs) kits lacked sensitivity, but interestingly, the combined use of both kits had a sensitivity and specificity of 100%. qPCRs detected SRLV infection before seroconversion at two weeks post infection and showed a specificity of 100%. Sensitivity however remained suboptimal at 85%. These results allow to propose a faster and cheaper diagnostic testing strategy for Belgium by combining a first ELISA screening, followed by confirmation of positive samples in AGID and/or a second ELISA. Since genotypes A and B strains are predominant in many countries, these results are interesting for other countries implementing SRLV control programs.

摘要

各国依赖良好的诊断检测和适当的检测方案来对抗具有经济重要性的小反刍动物慢病毒(SRLV)感染。我们使用大量具有代表性的比利时现场样本和实验感染绵羊和山羊的样本,对七种市售血清学检测方法和一种用于检测基因型 A 和 B 株的内部实时 PCR(qPCR)进行了广泛的比较分析。ELISA 通常表现良好,可在实验感染后三周内检测到血清转化。两种酶联免疫吸附试验(ELISAs)(Elitest 和 IDscreen 试剂盒)在两种物种中的敏感性(>96%)和特异性(>95%)最高,它们的联合使用可正确识别所有动物的感染状态。个别琼脂凝胶免疫扩散(AGID)试剂盒敏感性不足,但有趣的是,两个试剂盒的联合使用具有 100%的敏感性和特异性。qPCR 在感染后两周内可在血清转化之前检测到 SRLV 感染,特异性为 100%。然而,敏感性仍不理想,为 85%。这些结果允许通过组合第一个 ELISA 筛选,然后在 AGID 中确认阳性样本和/或第二个 ELISA,为比利时提出一种更快、更便宜的诊断检测策略。由于基因型 A 和 B 株在许多国家都很普遍,因此这些结果对其他实施 SRLV 控制计划的国家很有意义。

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