In:
Journal of Veterinary Research, Walter de Gruyter GmbH, Vol. 60, No. 2 ( 2016-6-1), p. 127-133
Kurzfassung:
Introduction: A real-time RT-PCR method for identification and quantification of porcine epidemic diarrhoea virus (PEDV) strains in China was developed. Material and Methods: Based on the conserved sequence of the PEDV nucleocapsid (N) gene, a primer pair and probe were designed to establish a TaqMan-MGB real-time RT-PCR assay for quantitative detection of the virus. The sequence was cloned into the pMD18-T vector and a series of diluted recombinant plasmids were used to generate a standard curve with an R2 value of 0.999. Results: The developed quantitative PCR assay detected viral titres as low as 0.1 TCID 50 with high specificity and no cross-reaction with other porcine viruses (PoRV, TGEV, PRRSV, or CSFV). The intra-batch and inter-batch coefficients of variation were both less than 1%, which indicated good reproducibility. Thirty clinical diarrhoea samples obtained from pigs in Shanghai and Fujian were analysed using this quantitative PCR assay. Out of these samples, 93.3% were found to be PEDV positive. Conclusion: This approach is suitable for clinical sample identification and pathogenesis studies.
Materialart:
Online-Ressource
ISSN:
2450-8608
DOI:
10.1515/jvetres-2016-0018
Sprache:
Englisch
Verlag:
Walter de Gruyter GmbH
Publikationsdatum:
2016
ZDB Id:
2855010-9
SSG:
22
