Molecular Investigation of Small Ruminant Abortions Using a 10-Plex HRM-qPCR Technique: A Novel Approach in Routine Diagnostics

oleh: Ioannis Gouvias, Marios Lysitsas, Apostolos Batsidis, Sonia Malefaki, Dimitra Bitchava, Anna Tsara, Emilija Nickovic, Ilias Bouzalas, Eleni Malissiova, Raphaël Guatteo, George Valiakos

Format: Article
Diterbitkan: MDPI AG 2024-08-01

Deskripsi

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 <i>Coxiella burnetii</i> and <i>Chlamydophila</i> 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. <i>Toxoplasma gondii</i>, <i>Salmonella</i> spp., <i>Brucella</i> spp., <i>Anaplasma phagocytophilum</i>, <i>Campylobacter fetus,</i> and <i>Neospora caninum</i> 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.