Preparation of milk samples for PCR analysis using a rapid filtration technique.
J Appl Microbiol
; 96(6): 1342-6, 2004.
Article
en En
| MEDLINE
| ID: mdl-15139927
AIMS: To investigate the usefulness of a straightforward filtration method for the isolation of Escherichia coli O157:H7 contaminants from milk for PCR detection. METHODS AND RESULTS: Escherichia coli O157:H7 is grown in milk and enriched in Luria-Bertani (LB) medium. Samples are filtered through a 0.45-microm pore membrane. The membrane is immersed in 200-microl lysis buffer and incubated at 95 degrees C for 10 min to release bacterial DNA for subsequent PCR detection. Under current conditions, the overall duration from filtration to PCR-ready DNA generation is <20 min, and the detection level for PCR was as low as 10 CFU of bacteria in 1 ml of milk. CONCLUSION: Bacterial contaminants of milk can be concentrated and isolated by a simple, one-step filtration and their DNA can be released for subsequent PCR detection by heating the filter membrane at 95 degrees C for 10 min. SIGNIFICANCE AND IMPACT OF THE STUDY: The simplicity of this method allows inexpensive, high throughput automation that meets the demands of modern food hygiene monitoring.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Escherichia coli O157
/
Leche
/
Microbiología de Alimentos
Tipo de estudio:
Evaluation_studies
Límite:
Animals
Idioma:
En
Revista:
J Appl Microbiol
Asunto de la revista:
MICROBIOLOGIA
Año:
2004
Tipo del documento:
Article
País de afiliación:
Taiwán
Pais de publicación:
Reino Unido