Evidence of Two Cocirculating Canine Distemper Virus Strains in Mesocarnivores From Northern Colorado, Usa

Abstract

Canine distemper virus (CDV) is a highly contagious pathogen that principally infects wildlife and domestic carnivores. Peridomestic species such as raccoons ( Procyon lotor) experience outbreaks with high mortality. Clinical signs of infection include anorexia, fever, respiratory infection, and neurologic complications. Although not zoonotic, CDV poses a high risk to unvaccinated domestic animals and the conservation of endangered species. During 2013-16, we opportunistically collected wild and domestic carnivore specimens through a rabies surveillance program in northern Colorado, US. Brainstem and cerebellar tissue samples were independently tested for rabies and CDV by fluorescent antibody test. We tested a total of 478 animals for CDV, comprised of 10 wild and domestic carnivore species. A total of 15% (72/478) of all animals sampled tested positive for CDV, consisting of 24% (71/300) of raccoons and 4% (1/26) of coyotes ( Canis latrans), but coinfection with rabies virus was not observed among CDV-positive animals. We extracted RNA from positive tissues, and a reverse-transcription PCR was used to create complementary DNA. We amplified and sequenced the hemagglutinin gene from 60 CDV-positive tissues, and a median joining network and maximum likelihood phylogenetic tree revealed two major lineages among samples. Phylogenetic analysis indicated that our sequences were most similar to the America-2 ( n=55) and the America-3 ( n=5) CDV lineages circulating in North America. Our results indicated two distinct and distantly related clades of CDV overlapping geographically and temporally among raccoon populations in northern Colorado.

Randomly primed, strand-switching, MinION-based sequencing for the detection and characterization of cultured RNA viruses

“…It is possible that the slightly lower percent identity for the swine IAV is a result of relative paucity of sequence data available for 2019 swine IAV at this time. The CDV America II lineage, the classification of CDV in our study, is a common lineage found in North American wildlife, 54 and is consistent with the collection of this sample from a raccoon in Kentucky. The sequences for IBV and PRRSV were highly similar to the known sequences of those isolates.…”

Randomly primed, strand-switching MinION-based sequencing for the detection and characterization of cultured RNA viruses

“…It is possible that the slightly lower percent identity for the swine-origin influenza is due to a relative paucity of sequence data available for 2019 swine-origin IAV at this time. The CDV America II lineage, the classification of CDV in this study, is a common lineage found in North American wildlife 42 and is consistent with the collection of this sample from a raccoon in Kentucky. The sequences for IBV and PRRSV were highly similar to the known sequences of those isolates.…”

Canine morbillivirus (CDV): a review on current status, emergence and the diagnostics

“…Virus detection a. Virus isolation (MDCK, Vero cells-SLAM B95a, etc) Virus Gold standard test Highly sensitive, helpful for generating virus repository (Require live virus titre, specific cell line and cell culture facilities) [ 7 ] [ 107 ] [ 120 ] Direct ELISA CDV antigen Detects antigen in serum [ 119 ] b. Antigen detection Sandwich ELISA H protein High specificity Detection and quantitation [ 91 ] F protein Efficient in field application with fecal and serum samples [ 148 ] Sandwich dot ELISA Virus Epidemiological surveillance [ 65 ] LFA F protein Practically applicable in the field for quick diagnosis [ 1 ] c. Nucleic acid detection RT-PCR N gene Standard laboratory test [ 39 ] One-step nested-RT-PCR N gene 100-fold sensitivity than RT-PCR and nested PCR [ 56 ] Double step real time-RT-PCR N gene Highly sensitive and specific Quantitate viral load in clinical samples [ 34 ] One-step real-time RT-PCR -- To study viral replication and kinetics of viral RNA load in infection …”