Detection of canine distemper virus nucleocapsid protein gene in canine peripheral blood mononuclear cells by RT-PCR.

For a rapid diagnosis of canine distemper virus (CDV) infection, the reverse transcription-PCR (RT-PCR) was carried out to detect CDV nucleoprotein (NP) gene from canine peripheral blood mononuclear cells (PBMCs). Two sets of primers were targeted to two regions of NP gene of CDV Onderstepoort strain. The NP gene fragments were well amplified by the RT-PCR in each of the RNA extracts from Vero cells infected with 6 laboratory strains of CDV including Onderstepoort strain, and from PBMCs of a dog experimentally infected with CDV. The amplified NP gene was detected in 17 of 32 samples from dogs which were clinically suspected for CDV infection at veterinary hospitals. No RT-PCR product was found in 52 samples from healthy dogs including 40 specific pathogen free beagles vaccinated with an attenuated live virus-vaccine for CDV and 12 stray dogs. The RT-PCR provides a fast, sensitive, and supplementary method for the diagnosis of CDV infection in dogs.

[1]  C. Kai,et al.  Use of B95a cells for isolation of canine distemper virus from clinical cases. , 1993, The Journal of veterinary medical science.

[2]  T. Kuiken,et al.  Characterization of morbilliviruses isolated from dolphins and porpoises in Europe. , 1993, The Journal of general virology.

[3]  V. ter meulen,et al.  Constant and variable regions of measles virus proteins encoded by the nucleocapsid and phosphoprotein genes derived from lytic and persistent viruses. , 1992, Virology.

[4]  E. Norrby,et al.  Sequence analysis of the genes encoding the nucleocapsid protein and phosphoprotein (P) of phocid distemper virus, and editing of the P gene transcript. , 1992, The Journal of general virology.

[5]  T. Harder,et al.  Detection of phocid distemper virus RNA in seal tissues using slot hybridization and the polymerase chain reaction amplification assay: genetic evidence that the virus is distinct from canine distemper virus. , 1991, The Journal of general virology.

[6]  A. Sugiura,et al.  B95a, a marmoset lymphoblastoid cell line, as a sensitive host for rinderpest virus. , 1991, The Journal of general virology.

[7]  D. Gajdusek,et al.  Detection of measles virus genomic sequences in sspe brain tissue by the polymerase chain reaction , 1990, Journal of medical virology.

[8]  A. Sugiura,et al.  Marmoset lymphoblastoid cells as a sensitive host for isolation of measles virus , 1990, Journal of virology.

[9]  G. Allan,et al.  Confirmation of cause of recent seal deaths , 1988, Nature.

[10]  S. Konishi,et al.  Kennel cough complexに関する研究 : わが国における流行の確認と病原の解析 , 1988 .

[11]  T. Wild,et al.  Antigenic analysis of African measles virus field isolates: identification and localisation of one conserved and two variable epitope sites on the NP protein. , 1988, Virus research.

[12]  M. Billeter,et al.  Multiple viral mutations rather than host factors cause defective measles virus gene expression in a subacute sclerosing panencephalitis cell line , 1988, Journal of virology.

[13]  K. Mullis,et al.  Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. , 1988, Science.

[14]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[15]  Y. Yoshikawa,et al.  Comparison of biological and molecular properties among canine distemper virus strains. , 1986, Nihon juigaku zasshi. The Japanese journal of veterinary science.

[16]  W. Bellini,et al.  Sequence homology within the morbilliviruses , 1985, Journal of virology.

[17]  S. Krakowka,et al.  Serologic response to canine distemper viral antigens in gnotobiotic dogs infected with canine distemper virus. , 1975, The Journal of infectious diseases.

[18]  R. L. Ott,et al.  Etiologic Studies of Old Dog Encephalitis , 1971, Veterinary pathology.

[19]  B. Mahy,et al.  The Molecular Biology of the Morbilliviruses , 1991 .

[20]  Gillespie Jh,et al.  Canine Distemper Virus , 1972, Virology Monographs / Die Virusforschung in Einzeldarstellungen.