Optimización de la Reacción en Cadena de la Polimerasa para la Detección del gen B1 de T. gondii
Optimization of Polymerase Chain Reaction for the detection of the B1 gene of T. gondii
Optimization of Polymerase Chain Reaction for the detection of the B1 gene of T. gondii
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Optimizar las condiciones de amplificación del gen B1 (35 copias en el genoma) para la detección de ADN de T.gondii en casos probables de toxoplasmosis cerebral. Materiales y métodos: Se realizó extracción de ADN a partir de exudado peritoneal de ratones inoculados con la cepa RH de T. gondii obteniendo 17 mL con una concentración inicial de 1x107 parásitos/mL. Se optimizaron las condiciones de PCR del gen B1. Resultados: Se obtuvo amplificación de un fragmento de 132 pb a partir de ADN obtenido de diluciones seriadas desde 1x106 a 1x10-1 parásitos por mL, estableciéndose un límite de detección de 1 taquizoíto de T. gondii. Objective: This study aimed at optimizing the amplification conditions of the B1 gene (35 copies in the genome) of T. gondii in cases of possible brain toxoplasmosis. Materials and methods: DNA was extracted from the peritoneal exudate of mice inoculated with the RH strain of T. gondii. Total exudate volume was 17 mL with a concentration of 1x107 parasites/mL. PCR conditions for the B1 gene were further optimized.Results: Serial dilutions from 1x106 to 1x10-1 parasites/mL were needed for amplifying a fragment of 132 bp . This set the detection limit for 1 T. gondii tachyzoite.
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http://hemeroteca.unad.edu.co/index.php/nova/article/view/1044http://dx.doi.org/10.22490/24629448.1044
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