Infectious pathogens that cause abortions in ruminants result in major economic losses to the global livestock industry. Pathogens associated with abortion have a direct impact on animal and human health. The objective of this study was to develop and validate a quantitative PCR-based diagnostic panel for rapid and simultaneous detection of some important pathogens causing abortion in sheep, goats, and cattle. For this purpose, standard curves were constructed using standard controls of the pathogens (Akabane, Peste des Petits Ruminants, Bluetongue, Bovine Viral Diarrhea, Border Disease, Bovine Herpesvirus, Schmallenberg viruses, Coxiella burnetii, Listeria monocytogenes, Chlamydophila abortus, Toxoplasma gondii, and Neospora caninum) to develop a one-run qPCR diagnostic panel. These curves were used to determine linearity, reliability, efficacy, and sensitivity of the test panel. Thus, the assay verification was completed, and the results were evaluated. The developed panel was verified on ovine (n=156), caprine (n=17), and bovine (n=15) fetus samples from the field, and causative agents of abortion in the region were identified. The limit of detection (LOD), amplification efficiency (E), regression (R²), and slope (M) values were determined for optimization and verification studies. The LOD for one-run qPCR assays ranged from 0.45 to 3.24 log₁₀ DNA copies/reaction and 1.33 to 4.64 log₁₀ RNA copies/reaction. The dynamic range for each one-run qPCR assay covered more than five orders of magnitude, and E values ranged between 90 and 100%; R² value for all test designs was 0.99; M values also ranged between -3.10 and -3.60. In conclusion, this diagnostic system, which has completed verification studies, can be actively used in routine diagnosis of domestic ruminant abortions, can analyze clinical sample assays within a few hours and can detect all target pathogens simultaneously.

To Cite This Article: Dogan M, Oz ME and Akbaba S 2025. One-Run qPCR assays for identification of domestic ruminant abortion: verification and application process. Pak Vet J, 45(2): 553-565. http://dx.doi.org/10.29261/pakvetj/2025.181