ABSTRACT
Aves polyomavirus 1, psittacine beak and feather disease virus, and psittacid herpesvirus 1 are important pathogens of psittacine birds with the potential to cause substantial morbidity and mortality. Using publically available nucleotide sequences, we developed and validated a triplex real-time PCR (rtPCR) assay to rapidly detect these 3 viruses. The assay had high analytical sensitivity, detecting <6 copies of viral DNA per reaction, and 100% analytical specificity, showing no cross-reactivity with 59 other animal pathogens. Archived formalin-fixed, paraffin-embedded tissues from psittacine birds diagnosed at postmortem as infected with each of the viruses as well as virus-negative birds were used to validate the utility of the assay. Birds were selected for the positive cohort if they showed histologic evidence of infection (i.e., characteristic inclusion bodies in tissues); birds in the negative cohort had final diagnoses unrelated to the pathogens of interest. The triplex rtPCR assay confirmed 98% of histopathology-positive cases, and also identified subclinical infections that were not observed by histologic examination, including coinfections. Birds that tested positive only by rtPCR had significantly higher cycle threshold values compared to those with histologic evidence of infection. Positive, negative, and overall percentage agreements as well as the kappa statistic between the results of the assay and histopathology were high, demonstrating the usefulness of the assay as a tool to confirm disease diagnoses, and to improve detection of subclinical infections.
Subject(s)
Bird Diseases/diagnosis , DNA Virus Infections/veterinary , DNA Viruses/isolation & purification , Herpesviridae/isolation & purification , Multiplex Polymerase Chain Reaction/veterinary , Psittaciformes/virology , Alphaherpesvirinae/genetics , Alphaherpesvirinae/isolation & purification , Animals , Bird Diseases/virology , Circoviridae Infections/diagnosis , Circoviridae Infections/veterinary , Circoviridae Infections/virology , Circovirus/genetics , Circovirus/isolation & purification , DNA Virus Infections/diagnosis , DNA Virus Infections/virology , DNA Viruses/genetics , DNA, Viral , Herpesviridae/genetics , Parrots/virology , Polyomaviridae/genetics , Polyomaviridae/isolation & purification , Polyomavirus/genetics , Polyomavirus/isolation & purification , Real-Time Polymerase Chain Reaction/veterinaryABSTRACT
In January, 2014, increased mortality was reported in piglets with acute diarrhea on an Ontario farm. Villus atrophy in affected piglets was confined to the small intestine. Samples of colon content were PCR-positive for porcine epidemic diarrhea virus (PEDV). Other laboratory tests did not detect significant pathogens, confirming this was the first case of PED in Canada.
Premier cas de diarrhée épidémique porcine au Canada. En janvier 2014, une mortalité accrue a été signalée chez des porcelets atteints de diarrhée aiguë dans une ferme de l'Ontario. L'atrophie des villosités chez les porcelets touchés a été confinée au petit intestin. Des échantillons du contenu du côlon étaient positifs par RCP pour le virus de la diarrhée épidémique porcine (VDEP). D'autres tests de laboratoire n'ont pas détecté d'agents pathogènes importants, ce qui confirme qu'il s'agit du premier cas de DEP au Canada.(Traduit par Isabelle Vallières).
