A multi-screening Fast qPCR approach to the identification of abortive agents in ruminants

• Published in: Journal of microbiological methods Vol. 148; pp. 12 - 17
• Main Authors: Sebastiani, Carla, Curcio, Ludovica, Ciullo, Marcella, Cruciani, Deborah, Crotti, Silvia, Pesca, Cristina, Torricelli, Martina, Sebastianelli, Martina, Felici, Andrea, Biagetti, Massimo
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.05.2018
• Subjects: Abortion, Septic - diagnosis; Abortion, Septic - veterinary; Abortive agents; Animals; Fast qPCR; Female; Mass Screening - methods; Molecular Diagnostic Techniques - methods; Multi-screening; Multiplex Polymerase Chain Reaction - methods; Pregnancy; Pregnancy Complications, Infectious - diagnosis; Pregnancy Complications, Infectious - veterinary; Rapid method; Real-Time Polymerase Chain Reaction - methods; Ruminants; Sensitivity and Specificity; Rapid method; Fast qPCR; Abortive agents; Ruminants; Multi-screening
• ISSN: 0167-7012; 1872-8359
• DOI: 10.1016/j.mimet.2018.03.009

Abortion in ruminants represents an important economic concern for farmers. Microbial agents, such as Brucella spp., Chlamydia spp., Coxiella burnetii, Leptospira spp., Neospora caninum, Salmonella spp. and Toxoplasma gondii, are among the main infectious causes of abortion and require rapid and reliable diagnosis. This study describes the development of a multi-screening assay using Fast Real-Time PCR (Fast qPCR) that allows, in a single test, the simultaneous identification of the above-mentioned abortive agents. This multi-screening approach is characterized by a mean diagnostic sensitivity and specificity of 100% and 97%, respectively; it has a limit of detection (LOD) ranging from 5 × 103 to 4 × 104 genomic copies/g of tissue and a very good concordance with traditional end-point PCR assays used in routine diagnostic activity. The proposed method represents a rapid approach to the simultaneous detection of the main abortive agents in ruminants that allows to make an accurate diagnosis and to set up appropriate control measures in a short period of time. [Display omitted] •A Multi-screening assay for the detection of 7 ruminant abortive agents was developed.•Fast qPCR quickly and reliably identifies ruminant abortive agents.•Limit of Detection (LOD) of the method ranging from 5 × 103 to 4 × 104 genomic copies/g of tissue•The assay has a diagnostic sensitivity of 100% and a diagnostic specificity of 97%.•The assay is modular and open to further development.