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Development of a SYBR Green I based duplex real-time PCR for detection of bovine herpesvirus-1 in semen.
- Source :
-
Journal of Virological Methods . Nov2014, Vol. 208, p6-10. 5p. - Publication Year :
- 2014
-
Abstract
- Bovine herpesvirus-1 (BoHV-1) is a viral pathogen found in infected bull semen, which is transmitted to inseminated cows by artificial insemination. BoHV-1 infection can cause reproductive disorders leading to significant economic loss to cattle industry. To detect BoHV-1 in semen, in this study, a SYBR Green I based duplex real-time PCR was developed. The assay included primers from BoHV-1 glycoprotein C (gC) and bovine growth hormone (bGH) genes for simultaneous detection in single tube. The result was interpreted by analysing melting temperature ( T m ) peaks obtained after melt curve analysis of the amplified products at the end of reaction. The T m peaks for BoHV-1-gC indicated presence of BoHV-1 while the bGH peak indicated reaction without inhibition. The sensitivity of the assay was to detect ten BoHV-1 genome copies per reaction. The analytical sensitivity was to detect 0.21 TCID 50 infectious BoHV-1 in spiked semen. The assay was validated with 80 semen samples collected from breeding bulls. The diagnostic sensitivity and specificity of the assay was 100% with OIE recommended TaqMan probe based real-time PCR. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 01660934
- Volume :
- 208
- Database :
- Academic Search Index
- Journal :
- Journal of Virological Methods
- Publication Type :
- Academic Journal
- Accession number :
- 98480143
- Full Text :
- https://doi.org/10.1016/j.jviromet.2014.07.027