Validation of a real-time PCR assay for the detection of bovine herpesvirus 1 in bovine semen

Wang, Jianning; O’Keefe, John; Orr, Della; Loth, Leo; Banks, M.; Wakeley, Philip R.; West, Donna; Card, Roderick M.; Ibata, G.; Maanen, Kees van; Thorén, Peter; Isaksson, Mats; Kerkhofs, Pierre

doi:10.1016/j.jviromet.2007.04.002

Cited by 52 publications

(47 citation statements)

References 21 publications

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“…Our finding on real time assay is in agreement with recent finding and was with better sensitivity/detection limits compared to the assays that have been described previously [8,29,30]. The assay can be efficiently used for diagnosis of BoHV-1 in clinical samples with special reference to semen samples which contain very low levels of virus and because if unchecked, an infected bull can be source of infection to thousands of female animals.…”

Section: Discussionsupporting

confidence: 91%

“…This is the smallest amplicon described for BoHV-1 diagnosis using real time PCR assay, as on date. Prior to our work Abril and colleagues [1], have reported the first set of primers and probe with an amplicon size of 97 bp for detection of BoHV-1 and these were validated by Wang et al [29]. The real time PCR developed in this study had a detection limit at 0.001 TCID 50 /0.1 ml of BoHV-1 in tissue culture supernatants .…”

Section: Discussionmentioning

confidence: 78%

“…This ultra low detection limit achieved during the study is attributed to the primers and probe targeted to amplify a very short, conserved region of BoHV-1. Wang et al [29,30] have reported a detection limit of 0.36 TCID 50 /0.1 ml of BoHV-1 in tissue culture supernatants.…”

Section: Discussionmentioning

confidence: 99%

“…This is significant, since shorter distances between the forward primer, the probe and reverse primer in real time assays increases the specificity of the assay [29]. This is the smallest amplicon described for BoHV-1 diagnosis using real time PCR assay, as on date.…”

Section: Discussionmentioning

confidence: 99%

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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

et al. 2012

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This study describes development of a TaqMan probe based real time PCR assay that can detect BoHV-1 of as low as 0.001 TCID 50 /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 C99 %.

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Section: Discussionsupporting

confidence: 91%

Section: Discussionmentioning

confidence: 78%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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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

et al. 2012

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“…The differential diagnosis process continued with the following molecular tests: BVDV, 6,25 VSV (protocol used for detection subtypes Alagoas, Indiana, New Jersey, and Cocal), 29 OvHV-2 and BTV, 18,30 orthopoxvirus (OPV), 13 PPV, 37 and BoHV-1. 33 All PCR assays were accompanied by blank controls to evaluate contamination.…”

Section: Molecular Analysismentioning

confidence: 99%

Detection of multiple viral infections in cattle and buffalo with suspected vesicular disease in Brazil

Laguardia-Nascimento¹,

Sales²,

Gasparini³

et al. 2016

J VET Diagn Invest

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Abstract. Vesicular diseases are of high importance for livestock, primarily because of foot-and-mouth disease (FMD), which is a high-morbidity disease that generates direct losses caused by low milk production, weight loss, and indirect losses because of the need for sanitary barriers. Other vesicular diseases are also of importance for livestock because of direct impacts or because their clinical signs may be confused with those of FMD. We report herein the detection of multiple infections in cattle with suspected vesicular disease in the Brazilian states of Amazonas (AM), Mato Grosso (MT), and Roraima. Thirtyseven epithelial samples from cattle and 1 sample from a buffalo were sent to the laboratory for testing for FMDV and similar disease agents. All samples from MT were positive for parapoxvirus (Pseudocowpox virus and Bovine papular stomatitis virus). In addition, 3 samples were positive for Bluetongue virus, and 5 samples were positive for Bovine herpesvirus 1. Among these samples, 1 was positive for all of these 3 agents. Only 2 samples from AM were negative for parapoxvirus. The molecular tests conducted in this study detected multiple infections, with a high prevalence of parapoxvirus.

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Infectious Agents

Tibary

2014

Bovine Reproduction

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Validation of a real-time PCR assay for the detection of bovine herpesvirus 1 in bovine semen

Cited by 52 publications

References 21 publications

Development of a Probe Based Real Time PCR Assay for Detection of Bovine Herpes Virus-1 in Semen and Other Clinical Samples

Development of a Probe Based Real Time PCR Assay for Detection of Bovine Herpes Virus-1 in Semen and Other Clinical Samples

Detection of multiple viral infections in cattle and buffalo with suspected vesicular disease in Brazil

Infectious Agents

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