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Development of a probe based real time PCR assay for detection of bovine herpes virus-1 in semen and other clinical samples.

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Title Development of a probe based real time PCR assay for detection of bovine herpes virus-1 in semen and other clinical samples.
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Creator Chandranaik BM
Rathnamma D
Patil SS
Kovi RC
Dhawan J
Ranganatha S
Isloor S
Renukaprasad C
Prabhudas K
 
Subject Bovine herpes virus-1
Indirect ELISA
Real time PCR
Virus isolation
 
Description Not Available
This study describes development of a TaqMan probe based real time PCR assay that can detect BoHV-1 of as low as 0.001 TCID50/0.1 ml in clinical samples, its comparative evaluation with indirect ELISA and virus isolation for detection of Bovine herpes virus-1 (BoHV-1) in semen and swab clinical samples. For this study, we collected samples from 212 animals (cattle and buffaloes) comprising 91 bulls and 121 females. Avidin-biotin ELISA employed on serum samples from 212 animals revealed 74 as seropositive for BoHV-1. On inoculation of semen/swabs on MDBK cell line, nine samples yielded cytopathic changes characteristic of herpes viruses. The isolates were confirmed by VNT and a conventional PCR. A real time PCR assay was standardised by designing a new set of TaqMan probe and primers targeting a 71 bp region on gB gene of the virus. The assay detected viral antigen in 21 seropositive and 14 seronegative animals, emphasizing the relevance of serology in BoHV-1 diagnosis, particularly in breeding stations. Further, real time PCR assay was 100 % sensitive and 87.19 % specific compared to virus isolation in detection of the BoHV-1 in clinical samples. The assay was validated at reputed national laboratories, with a sensitivity of ≥99 %.
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Date 2018-10-29T11:18:08Z
2018-10-29T11:18:08Z
2013-06-01
 
Type Research Paper
 
Identifier Not Available
Not Available
http://krishi.icar.gov.in/jspui/handle/123456789/8583
 
Language English
 
Relation Not Available;
 
Publisher Pubmed