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

Autor: Sebastiani C; R&D Department, Istituto Zooprofilattico Sperimentale dell'Umbria e delle Marche - Togo Rosati (IZSUM), Via Salvemini 1, 06126 Perugia, Italy., Curcio L; R&D Department, Istituto Zooprofilattico Sperimentale dell'Umbria e delle Marche - Togo Rosati (IZSUM), Via Salvemini 1, 06126 Perugia, Italy., Ciullo M; R&D Department, Istituto Zooprofilattico Sperimentale dell'Umbria e delle Marche - Togo Rosati (IZSUM), Via Salvemini 1, 06126 Perugia, Italy., Cruciani D; Diagnostic Laboratory, Istituto Zooprofilattico Sperimentale dell'Umbria e delle Marche - Togo Rosati (IZSUM), Via Salvemini 1, 06126 Perugia, Italy., Crotti S; Diagnostic Laboratory, Istituto Zooprofilattico Sperimentale dell'Umbria e delle Marche - Togo Rosati (IZSUM), Via Salvemini 1, 06126 Perugia, Italy., Pesca C; Diagnostic Laboratory, Istituto Zooprofilattico Sperimentale dell'Umbria e delle Marche - Togo Rosati (IZSUM), Via Salvemini 1, 06126 Perugia, Italy., Torricelli M; R&D Department, Istituto Zooprofilattico Sperimentale dell'Umbria e delle Marche - Togo Rosati (IZSUM), Via Salvemini 1, 06126 Perugia, Italy., Sebastianelli M; R&D Department, Istituto Zooprofilattico Sperimentale dell'Umbria e delle Marche - Togo Rosati (IZSUM), Via Salvemini 1, 06126 Perugia, Italy., Felici A; Epidemiology Unit, Istituto Zooprofilattico Sperimentale dell'Umbria e delle Marche - Togo Rosati (IZSUM), Via Salvemini 1, 06126 Perugia, Italy., Biagetti M; R&D Department, Istituto Zooprofilattico Sperimentale dell'Umbria e delle Marche - Togo Rosati (IZSUM), Via Salvemini 1, 06126 Perugia, Italy. Electronic address: m.biagetti@izsum.it.
Jazyk: angličtina
Zdroj: Journal of microbiological methods [J Microbiol Methods] 2018 May; Vol. 148, pp. 12-17. Date of Electronic Publication: 2018 Mar 21.
DOI: 10.1016/j.mimet.2018.03.009
Abstrakt: 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 × 10 3 to 4 × 10 4 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.
(Copyright © 2018 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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