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

J Microbiol Methods. 2018 May:148:12-17. doi: 10.1016/j.mimet.2018.03.009. Epub 2018 Mar 21.

Abstract

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.

Keywords: Abortive agents; Fast qPCR; Multi-screening; Rapid method; Ruminants.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Abortion, Septic / diagnosis
  • Abortion, Septic / veterinary*
  • Animals
  • Female
  • Mass Screening / methods*
  • Molecular Diagnostic Techniques / methods*
  • Multiplex Polymerase Chain Reaction / methods*
  • Pregnancy
  • Pregnancy Complications, Infectious / diagnosis
  • Pregnancy Complications, Infectious / veterinary*
  • Real-Time Polymerase Chain Reaction / methods*
  • Ruminants*
  • Sensitivity and Specificity