Desarrollo de una PCR en tiempo real multiplex para la detección de BoHV-1 y BVDV en semen bovino criopreservado
Palabras clave:
inseminación artificial, TaqMan, bioseguridadResumen
En México, la inseminación artificial es una de las principales biotecnologías reproductivas en la ganadería bovina, empleada para incrementar la productividad y el mejoramiento genético del hato. No obstante, gran parte del semen utilizado se procesa a nivel de campo sin un control sanitario estricto, lo que incrementa el riesgo de diseminación de agentes infecciosos como el virus de la diarrea viral bovina (BVDV) y el herpesvirus bovino tipo 1 (BoHV-1). Para fortalecer la detección de estos patógenos, se desarrolló una PCR en tiempo real multiplex con sondas TaqMan, capaz de identificar simultáneamente ambos virus en semen bovino criopreservado. Se emplearon controles positivos consistentes en virus de referencia replicados en células MDBK y, como controles negativos, sobrenadantes de células no infectadas y semen libre de los virus. Los cebadores y sondas se diseñaron a partir de secuencias depositadas en GenBank: el gen E2 para BVDV y la glicoproteína gB para BoHV-1; adicionalmente, se utilizó el gen de ?-actina de Bos taurus como constitutivo. Los ensayos demostraron amplificación específica en pruebas de punto final y resultados satisfactorios en tiempo real. Los hallazgos obtenidos permiten establecer una prueba diagnóstica multiplex que contribuirá a la bioseguridad en programas de inseminación artificial, con la finalidad de minimizar el riesgo de transmisión de BVDV y BoHV-1 a través del semen.
https://doi.org/10.21929/abanicomicrobiano/2025.6
2025-6
https://www.youtube.com/watch?v=FmFfdKDZACY
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