Gouvias Ioannis, Lysitsas Marios, Batsidis Apostolos, Malefaki Sonia, Bitchava Dimitra, Tsara Anna, Nickovic Emilija, Bouzalas Ilias, Malissiova Eleni, Guatteo Raphaël, Valiakos George
Faculty of Veterinary Science, University of Thessaly, 43100 Karditsa, Greece.
Department of Mathematics, University of Ioannina, 45110 Ioannina, Greece.
Microorganisms. 2024 Aug 14;12(8):1675. doi: 10.3390/microorganisms12081675.
The objective of this study was to apply and preliminarily evaluate a High-Resolution Melting (HRM) analysis technique coupled with qPCR, that allows the simultaneous detection of 10 different ruminant abortogenic pathogens, for investigating abortions in sheep and goats throughout Greece. A total of 264 ovine and caprine vaginal swabs were obtained the week following the abortion from aborted females and analyzed using a commercially available kit (ID Gene™ Ruminant Abortion Multiplex HRM, Innovative Diagnostics). Results indicated a high prevalence of and spp., which were detected in 48.9% and 42.4% of the vaginal swabs, respectively. Results for these most commonly detected pathogens were compared with those of a well-established commercial qPCR kit, with near-perfect agreement. , spp., spp., , and were also identified, the two latter reported for the first time in the country in small ruminants. Mixed infections occurred in 35.6% of the animals examined. This technique allows for the simultaneous detection of many abortogenic pathogens in an accurate and cost-effective assay. Detection of uncommon or not previously reported pathogens in various cases indicates that their role in ovine and caprine abortions may be underestimated.
本研究的目的是应用并初步评估一种结合定量聚合酶链反应(qPCR)的高分辨率熔解曲线分析(HRM)技术,该技术可同时检测10种不同的反刍动物流产病原体,用于调查希腊各地绵羊和山羊的流产情况。在流产后的一周内,从流产母羊中采集了264份绵羊和山羊阴道拭子,并使用市售试剂盒(ID Gene™反刍动物流产多重HRM,创新诊断公司)进行分析。结果表明,支原体属和脲原体属的感染率很高,在阴道拭子中分别检测到48.9%和42.4%。将这些最常检测到的病原体的检测结果与一种成熟的商业qPCR试剂盒的结果进行比较,结果几乎完全一致。还鉴定出了嗜组织菌属、衣原体属、新孢子虫属、沙门氏菌属、李斯特菌属和布鲁氏菌属,后两者在该国首次在小型反刍动物中被报道。35.6%的受检动物发生了混合感染。该技术能够在一种准确且经济高效的检测方法中同时检测多种流产病原体。在各种病例中检测到不常见或以前未报道过的病原体,表明它们在绵羊和山羊流产中的作用可能被低估了。