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Jpn J Infect Dis ; 65(2): 111-6, 2012.
Artículo en Inglés | MEDLINE | ID: mdl-22446116

RESUMEN

We developed an enrichment medium for use with the loop-mediated isothermal amplification (LAMP) assay (enrichment media + LAMP assay) to quickly increase a very small number of Vibrio parahaemolyticus cells to the detection limit of the assay. Thirty-nine different enrichment media were prepared based on evaluating 12 potential ingredients. From our assessment of the 39 media, enrichment medium #36, which contained 2% sodium chloride, 1% proteose peptone no. 2, 0.1% trehalose, 0.5% α-ketoglutaric acid, 0.25% pyruvic acid, and 0.5% yeast extract (pH 8.6), was found to be most effective at enhancing the proliferation of V. parahaemolyticus during incubation for 3 h at 40ºC. We compared the detection limits of the LAMP assay, the enrichment medium #36 + LAMP assay, and the cultivation method using bacterial cell and spiked shrimp sample tests. The detection limits of the LAMP assay, the medium #36 + LAMP assay, and the cultivation method were 10(3), 10(0)-10(-1), and 10(-1) CFU ml(-1), respectively. Enrichment medium #36 promoted a 10(3)- to 10(4)-fold increase in the bacterial population, and the detection limit of the enrichment media + LAMP assay was the same as that of the cultivation method.


Asunto(s)
Técnicas Bacteriológicas/métodos , Medios de Cultivo/química , Técnicas de Amplificación de Ácido Nucleico/métodos , Vibriosis/diagnóstico , Vibrio parahaemolyticus/aislamiento & purificación , Animales , Humanos , Límite de Detección , Temperatura , Factores de Tiempo , Vibriosis/microbiología
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