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Detection and differentiation of porcine epidemic diarrhoea virus and transmissible gastroenteritis virus in clinical samples by multiplex RT-PCR.
- Source :
-
The Veterinary record [Vet Rec] 2000 May 27; Vol. 146 (22), pp. 637-40. - Publication Year :
- 2000
-
Abstract
- A multiplex reverse-transcriptase-PCR (RT-PCR) procedure was developed for the simultaneous detection of porcine epidemic diarrhoea virus (PEDV) and transmissible gastroenteritis virus (TGEV) in preweaning pigs with diarrhoea. The membrane gene of PEDV and the nucleocapsid gene of TGEV were chosen as targets. The PCR products of PEDV and TGEV had molecular sizes of 412 and 612 base pairs, respectively. Primers from PEDV did not react with any TGEV tested and vice versa. In addition, the primers did not react with other pig viruses. The multiplex RT-PCR was able to detect 10 tissue culture-infective doses 50 per cent (TCID50)/ml of PEDV or TGEV with each of the primer sets for PEDV and TGEV, respectively. The RNAS of PEDV and TGEV were detected directly in intestinal and faecal samples from pigs infected experimentally with either virus. The results of the assay correlated well with the results of virus isolation. None of the five control specimens was positive. PEDV was detected in 10 intestinal and nine faecal samples, and among the nine positive faecal samples two were culture-negative. TGEV was also detected in 10 intestinal and nine faecal samples, and among the nine positive faecal samples, three were culture-negative.
- Subjects :
- Animals
Animals, Newborn
Coronaviridae genetics
Coronaviridae Infections diagnosis
Coronaviridae Infections veterinary
Coronaviridae Infections virology
DNA Primers
Diagnosis, Differential
Diarrhea diagnosis
Diarrhea virology
Feces virology
Female
Gastroenteritis, Transmissible, of Swine diagnosis
Intestines virology
Male
Sensitivity and Specificity
Swine
Swine Diseases diagnosis
Swine Diseases virology
Transmissible gastroenteritis virus genetics
Coronaviridae isolation & purification
Diarrhea veterinary
Gastroenteritis, Transmissible, of Swine virology
Reverse Transcriptase Polymerase Chain Reaction veterinary
Transmissible gastroenteritis virus isolation & purification
Subjects
Details
- Language :
- English
- ISSN :
- 0042-4900
- Volume :
- 146
- Issue :
- 22
- Database :
- MEDLINE
- Journal :
- The Veterinary record
- Publication Type :
- Academic Journal
- Accession number :
- 10872784
- Full Text :
- https://doi.org/10.1136/vr.146.22.637