SYBR Green I Based Real-Time RT-PCR assay for the detection of bovine herpesvirus type 1

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Published: Dec 1, 2019
Keywords:
bovine herpesvirus type 1, real-time PCR, SYBR-Green I

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Laiyen García Delgado
Centro Nacional de Sanidad Agropecuaria
Roxana Marrero-Perera
Centro Nacional de Sanidad Agropecuaria
Liani Coronado Báez
Centro Nacional de Sanidad Agropecuaria
Yoandri Hinojosa López
Centro Nacional de Sanidad Agropecuaria
Ana María Acevedo
Centro Nacional de Sanidad Agropecuaria
Damarys Relova Vento
Centro Nacional de Sanidad Agropecuaria
Armando Vega Redondo
Centro Nacional de Sanidad Agropecuaria
Carmen Laura Perera González
Centro Nacional de Sanidad Agropecuaria

Abstract

Bovine herpesvirus type 1 (BoHV-1) infects the respiratory and genital tracts of cattle and several species of ruminants. The infection can result in different diseases such as infectious bovine rhinotracheitis, infectious pustular vulvovaginitis in females and infectious pustular balanoposthitis in males, with a severe economic impact on livestock production. The infection causes respiratory symptoms, conjunctivitis, abortions, enteritis, and fatal systemic infection in newborn calves. One of the most important characteristics of bovine herpesvirus type 1 is the establishment of a latent infection that can be reactivated under stress conditions or by treatment with high doses of corticosteroids. The present work was aimed at developing an SYBR Green I Based Real-Time RT-PCR assay for the detection of bovine herpesvirus type 1, by amplifying a fragment of the viral thymidine kinase gene. The parameters of the critical components of the PCR reaction as well as the thermodynamic conditions were established. Dissociation curve analysis showed that the amplification curves were specific (Tm = 93°C ± 1°C) and that there were no unspecific amplifications. High levels of specificity, repeatability and sensitivity were obtained, with a detection limit of 100.5 TCID50/mL. Diagnostic sensitivity was assessed on the basis of nasal exudates from experimental breeding animals, reactivation of latent infection and experimental dairy farms. The SYBR Green I Based Real-Time RT-PCR assay proved to be more sensitive for the diagnosis of BoHV-1 than the endpoint PCR assay and virus isolation.

Article Details

How to Cite
1.
García Delgado L, Marrero-Perera R, Coronado Báez L, Hinojosa López Y, Acevedo AM, Relova Vento D, Vega Redondo A, Perera González CL. SYBR Green I Based Real-Time RT-PCR assay for the detection of bovine herpesvirus type 1. Rev. Salud Anim. [Internet]. 2019 Dec. 1 [cited 2024 Nov. 22];41(3). Available from: https://revistas.censa.edu.cu/index.php/RSA/article/view/1040
Issue
Vol. 41 No. 3 (2019): septiembre-diciembre
Section
ARTÍCULOS ORIGINALES
National Center for Animal and Plant Health (CENSA)

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