Detection of canine distemper virus (CDV) through one step RT-PCR combined with nested PCR
Journal of Veterinary Science
; : 59-63, 2001.
Article
en En
| WPRIM
| ID: wpr-72517
Biblioteca responsable:
WPRO
ABSTRACT
A one step reverse transcription PCR (RT-PCR) combined nested PCR was set up to increase efficiency in the diagnosis of canine distemper virus (CDV) infection after developement of nested PCR. Two PCR primer sets were designed based on the sequence of nucleocapsid gene of CDV Onderstepoort strain. One-step RT-PCR with the outer primer pair was revealed to detect 10(2) PFU/ml. The sensitivity was increased hundredfold using the one-step RT-PCR combined with the nested PCR. Specificity of the PCR was also confirmed using other related canine virus and peripheral blood mononuclear cells (PBMC) and body secretes of healthy dogs. Of the 51 blood samples from dogs clinically suspected of CD, 45 samples were revealed as positive by one-step RT-PCR combined with nested PCR. However, only 15 samples were identified as positive with a single one step RT-PCR. Therefore approximately 60% increase in the efficiency of the diagnosis was observed by the combined method. These results suggested that one step RT-PCR combined with nested PCR could be a sensitive, specific, and practical method for diagnosis of CDV infection.
Palabras clave
Texto completo:
1
Índice:
WPRIM
Asunto principal:
ARN Viral
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Vacunas Atenuadas
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Vacunas Virales
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Reacción en Cadena de la Polimerasa
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Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
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Virus del Moquillo Canino
Tipo de estudio:
Diagnostic_studies
Límite:
Animals
Idioma:
En
Revista:
Journal of Veterinary Science
Año:
2001
Tipo del documento:
Article