Broadly Reactive Pan-Paramyxovirus Reverse Transcription Polymerase Chain Reaction and Sequence Analysis for the Detection of Canine Distemper Virus in a Case of Canine Meningoencephalitis of Unknown Etiology

Abstract

Abstract. Despite the immunologic protection associated with routine vaccination protocols, Canine distemper virus (CDV) remains an important pathogen of dogs. Antemortem diagnosis of systemic CDV infection may be made by reverse transcription polymerase chain reaction (RT-PCR) and/or immunohistochemical testing for CDV antigen; central nervous system infection often requires postmortem confirmation via histopathology and immunohistochemistry. An 8-month-old intact male French Bulldog previously vaccinated for CDV presented with multifocal neurologic signs. Based on clinical and postmortem findings, the dog's disease was categorized as a meningoencephalitis of unknown etiology. Broadly reactive, pan-paramyxovirus RT-PCR using consensusdegenerate hybrid oligonucleotide primers, combined with sequence analysis, identified CDV amplicons in the dog's brain. Immunohistochemistry confirmed the presence of CDV antigens, and a specific CDV RT-PCR based on the phosphoprotein gene identified a wild-type versus vaccinal virus strain. This case illustrates the utility of broadly reactive PCR and sequence analysis for the identification of pathogens in diseases with unknown etiology.

Viral Pathogen Detection by Metagenomics and Pan-Viral Group Polymerase Chain Reaction in Children With Pneumonia Lacking Identifiable Etiology

“…For children hospitalized with CAP, combined nasopharyngeal (NP) and oropharyngeal (OP) swabs were collected within 72 hours of hospital admission. Specimens were transferred into 3-mL universal transport media, refrigerated, and stored at −80°C within 24 , and respiratory syncytial virus) were detected by PCR of NP/OP swabs or serology of acute-and convalescent-phase serum (except for human rhinovirus and coronaviruses) [4]. Nasopharyngeal and OP swabs were also obtained from asymptomatic control subjects before elective surgery while in the operating room and tested for viral pathogens, C. pneumoniae, and M. pneumoniae [4].…”

“…They were designed using the consensus-degenerate hybrid oligonucleotide primer (CODEHOP) principle to conserved genes and regions [19,20] and had analytical sensitivities of 10-500 copies (RNA viruses) and 10-1000 copies (DNA viruses) per reaction. Samples (5 μl of total nucleic acid) were tested with broadly reactive PVG PCR for the following viral families/genera: Adenoviridae, Anelloviridae, Arenaviridae, Astroviridae, Bunyaviridae, Caliciviridae, Circoviridae, Coronaviridae, Flaviviridae (Flavivirus), Herpesviridae, Orthomyxoviridae (influenza viruses A, B, and C), Paramyxoviridae, Parvoviridae, Picornaviridae (enterovirus and parechovirus; primers do not target all human rhinoviruses), Polyomaviridae, Reoviridae (aquareovirus, orthoreovirus, orbivirus, rotavirus, and seadornavirus), Rhabdoviridae, and Togaviridae (alphavirus) [21][22][23][24][25][26][27][28]. The Picornaviridae PCR targets only a subset of human rhinoviruses.…”

Canine Distemper Spillover in Domestic Dogs from Urban Wildlife

“…27,32 The major vaccine strains were isolated in the 1930s and it is not known if they continue to circulate in nature as they have not been detected for many years. 33,34 Although CDV vaccine strains have not changed in the past 60 years, there is potential for newer antigenic variants of CDV to emerge around the world. 23 However, the current vaccines have largely provided adequate protection against clinical disease when properly administered to healthy domesticated dogs in this country.…”

Conventional and molecular diagnostic testing for the acute neurologic patient

“…Recently, the CDC has reported on a panel of 4 semi‐nested RT‐PCR assays that utilize CODEHOP primers for the diagnosis of paramyxovirus infections. We have utilized this pan‐paramyxovirus PCR to determine CDV as the etiology in an unusual case of necrotizing meningoencephalitis (NME) 65 …”