Development of a SYBR Green I based duplex real-time PCR for detection of bovine herpesvirus-1 in semen
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Title |
Development of a SYBR Green I based duplex real-time PCR for detection of bovine herpesvirus-1 in semen
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Creator |
Sachinkumar Shivajirao Pawar
P.K. Gupta |
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Subject |
SYBR Green I,duplex real-time PCR,bovine herpesvirus-1,semen
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Description |
Not Available
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 (Tm) peaks obtained after melt curve analysis of the amplified products at the end of reaction. The Tm 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 TCID50 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. |
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Date |
2021-05-14T14:09:24Z
2021-05-14T14:09:24Z 2014-11-1 |
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Type |
Research Paper
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Identifier |
Pawar SS, Meshram CD, Singh NK, Saini M, Mishra BP, Gupta PK. Development of a SYBR Green I based duplex real-time PCR for detection of bovine herpesvirus-1 in semen. J Virol Methods. 2014 Nov;208:6-10. doi: 10.1016/j.jviromet.2014.07.027. Epub 2014 Jul 29. PMID: 25078112.
Not Available 0166-0934 http://krishi.icar.gov.in/jspui/handle/123456789/47018 |
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Language |
English
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Publisher |
Not Available
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