A probe-based real-time PCR assay for the detection of Neospora caninum in clinical samples from cattle

Rhiannon Barry, Ruth H. Nissly, Willard Feria, Nagaraja Thirumalapura, Deepanker Tewari, Bhushan M. Jayarao, Suresh Varma Kuchipudi

Research output: Contribution to journalArticle

Abstract

Neospora caninum is an apicomplexan protozoan parasite that is a leading cause of abortion in cattle. Detection of parasite-specific DNA by PCR is a highly sensitive method for identifying the presence of N. caninum in a variety of tissues. We developed and validated a probe-based real-time PCR assay targeting the conserved Nc5 gene of N. caninum. Using N. caninum strain Nc-1 genomic DNA and a synthetic gene fragment as amplification standards, we determined the PCR amplification efficiency and the limit of detection to be 95.60% and 3 copies, respectively. Five pathogens frequently associated with bovine abortions, namely bovine viral diarrhea virus types I and II, bovine alphaherpesvirus-1, Chlamydia, and Leptospira, were tested to ensure analytical exclusivity. A total of 103 clinical samples from aborted fetuses were tested concurrently with a standard conventional PCR and the new probe-based real-time PCR assay. All tested samples showed 100% agreement between these two assays. In conclusion, the probe-based real-time PCR assay facilitates accurate and rapid detection of N. caninum from abortions in cattle.

Original languageEnglish (US)
Pages (from-to)2-6
Number of pages5
JournalVeterinary Parasitology
Volume269
DOIs
StatePublished - May 1 2019

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Neospora
Neospora caninum
Real-Time Polymerase Chain Reaction
quantitative polymerase chain reaction
abortion (animals)
cattle
assays
Polymerase Chain Reaction
Parasites
sampling
Aborted Fetus
Synthetic Genes
Bovine Viral Diarrhea Viruses
synthetic genes
parasites
Leptospira
Chlamydia
DNA
Bovine viral diarrhea virus
Protozoa

All Science Journal Classification (ASJC) codes

  • Parasitology
  • veterinary(all)

Cite this

Barry, Rhiannon ; Nissly, Ruth H. ; Feria, Willard ; Thirumalapura, Nagaraja ; Tewari, Deepanker ; Jayarao, Bhushan M. ; Kuchipudi, Suresh Varma. / A probe-based real-time PCR assay for the detection of Neospora caninum in clinical samples from cattle. In: Veterinary Parasitology. 2019 ; Vol. 269. pp. 2-6.
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A probe-based real-time PCR assay for the detection of Neospora caninum in clinical samples from cattle. / Barry, Rhiannon; Nissly, Ruth H.; Feria, Willard; Thirumalapura, Nagaraja; Tewari, Deepanker; Jayarao, Bhushan M.; Kuchipudi, Suresh Varma.

In: Veterinary Parasitology, Vol. 269, 01.05.2019, p. 2-6.

Research output: Contribution to journalArticle

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AU - Barry, Rhiannon

AU - Nissly, Ruth H.

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AU - Tewari, Deepanker

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AU - Kuchipudi, Suresh Varma

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