Comparison of reverse-transcription real-time PCR and immunohistochemistry for the detection of canine distemper virus infection in raccoons in Ontario, Canada

J Vet Diagn Invest. 2018 Mar;30(2):319-323. doi: 10.1177/1040638717751825. Epub 2017 Dec 28.

Abstract

Canine distemper virus (CDV) is a widespread morbillivirus that causes subclinical to fatal infections in domestic and wild carnivores. Raccoons ( Procyon lotor) are CDV reservoirs and suffer from associated disease. Aspects of pathogenesis may lead to difficulty in the interpretation of commonly used testing modalities, such as reverse-transcription real-time (RT-rt)PCR and immunohistochemistry (IHC). The reliance upon such tests is greater for wildlife, which are often submitted as carcasses with no clinical history. We compared CDV RT-rtPCR results to immunohistochemistry (the gold standard) in tissues from 74 raccoons. These tests had high kappa agreement (lymph node: 0.9335; lung: 0.8671) and a negative correlation between IHC score and threshold cycle (Ct) value for lymph node and lung (Spearman rank correlation coefficient [ rs] = -0.8555 and -0.8179, respectively; p < 0.00001). An RT-rtPCR Ct value of 30 in lung and lymph node with sensitivity and specificity of 92.3 and 92.6% and 86.8 and 96.4%, respectively, was suitable for determining CDV involvement. Conjunctival swabs provide an alternative for distemper diagnosis, as there was a strong correlation between Ct values of conjunctival swabs and tissues ( rs = -0.8498, p < 0.00001, n = 46). This information will aid in more efficient and accurate diagnoses in individuals, small-scale outbreaks, and epidemiologic investigations in wildlife.

Keywords: Detection test; distemper; morbillivirus; raccoon; wild animal.

Publication types

  • Comparative Study
  • Evaluation Study

MeSH terms

  • Animals
  • Animals, Wild
  • Disease Outbreaks / veterinary*
  • Distemper / epidemiology*
  • Distemper / virology
  • Distemper Virus, Canine / genetics
  • Distemper Virus, Canine / isolation & purification*
  • Female
  • Immunohistochemistry / veterinary
  • Male
  • Ontario / epidemiology
  • Raccoons*
  • Real-Time Polymerase Chain Reaction / veterinary
  • Sensitivity and Specificity