Detection of canine distemper virus (CDV) through one step RT-PCR combined with nested PCR.
- Author:
Yong Hwan KIM
;
Kyu Woan CHO
;
Hwa Young YOUN
;
Han Sang YOO
;
Hong Ryul HAN
- Publication Type:Original Article
- Keywords:
CDV;
RT-PCR;
nested PCR
- MeSH:
Animals;
Distemper Virus, Canine/genetics/*isolation & purification;
Dogs;
Polymerase Chain Reaction/*methods/*veterinary;
RNA, Viral/genetics/isolation & purification;
Reverse Transcriptase Polymerase Chain Reaction;
Vaccines, Attenuated;
Viral Vaccines
- From:Journal of Veterinary Science
2001;2(1):59-63
- CountryRepublic of Korea
- Language:English
-
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.
