La Reacción de Cadena de la Polimerasa como Herramienta Eficaz para la Detección de Mycoplasma en Cultivos Celulares
Resumen
Mycoplasma, bacterias sin pared celular, pueden contaminar cultivos celulares y comprometer la integridad de los experimentos biológicos, generando resultados erróneos si no se detectan. En este estudio se evaluaron dos técnicas para la detección de Mycoplasma en las líneas celulares SW613B3, HCT, HeLa, PC-3, PNT2 y RKO: la reacción en cadena de la polimerasa (PCR) y la tinción con 4’,6-diamidino-2-fenilindol (DAPI). La presencia de Mycoplasma se analizó mediante PCR, revelando contaminación en tres líneas: HCT, PC-3 y RKO, confirmada por la amplificación de productos de 270 pb. El análisis parcial del 16S rADN de HCT y PC-3 mostró una alta similitud (97,98–99,50%) con Mesomycoplasma hyorhinis. La tinción con DAPI permitió visualizar Mycoplasma en el citoplasma de las células contaminadas, apoyando los hallazgos moleculares. Asimismo, se evaluó la eficacia de un kit comercial y del antibiótico ciprofloxacino (10µg/mL) para eliminar la contaminación en las líneas PC-3 y PNT2, respectivamente. La ausencia de Mycoplasma se confirmó mediante PCR negativa tras el tratamiento. Ambos métodos de detección son efectivos para la identificación de Mycoplasma, destacando la PCR por su sensibilidad, especificidad y rapidez, por lo que se recomienda su uso en el control de calidad rutinario. Tanto el kit comercial como la ciprofloxacina demostraron ser estrategias prácticas y eficientes para la recuperación de cultivos contaminados.
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Derechos de autor 2026 Diana Flores Guevara, Ricardo Puglla Ganazhapa, Jackeline Elizabeth Guamán Hurtado, Paulo Herrera, Ana Paulina Arévalo Jaramillo
Esta obra está bajo licencia internacional Creative Commons Reconocimiento 4.0.
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