Journal of Rapid Methods & Automation in Microbiology

ISOLATION OF LISTERIA MONOCYTOGENES DNA FROM MEAT PRODUCTS FOR QUANTITATIVE DETECTION BY REAL-TIME PCR

Corresponding Author

DAVID RODRÍGUEZ-LÁZARO

Division of Veterinary Pathology, Infection and Immunity
Faculty of Medical and Veterinary Sciences
University of Bristol
BS40 5DU Langford, Bristol, United Kingdom

TEL: 44-0-117-331-9007; FAX: 44-0-117-928-9505; EMAIL: [email protected]Search for more papers by this author

MARTA HERNÁNDEZ,

MARTA HERNÁNDEZ

Laboratorio de Biologia Molecular
Subdirección de Investigación y Tecnología
Instituto Tecnológico Agrario de Castilla y León (ITACyL)
Carretera de Burgos, Km. 119
47.071 Valladolid, Spain

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DAVID RODRÍGUEZ-LÁZARO,

Corresponding Author

DAVID RODRÍGUEZ-LÁZARO

Division of Veterinary Pathology, Infection and Immunity
Faculty of Medical and Veterinary Sciences
University of Bristol
BS40 5DU Langford, Bristol, United Kingdom

TEL: 44-0-117-331-9007; FAX: 44-0-117-928-9505; EMAIL: [email protected]Search for more papers by this author

MARTA HERNÁNDEZ,

MARTA HERNÁNDEZ

Laboratorio de Biologia Molecular
Subdirección de Investigación y Tecnología
Instituto Tecnológico Agrario de Castilla y León (ITACyL)
Carretera de Burgos, Km. 119
47.071 Valladolid, Spain

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First published: 04 January 2007

https://doi.org/10.1111/j.1745-4581.2006.00058.x

Citations: 13

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ABSTRACT

We report a study of two different procedures for extraction of Listeria monocytogenes genomic DNA from cooked pork ham products, to be coupled to a specific real-time (RTi-) PCR assay, an open formula protocol based on the Chelex-100 resin and the Wizard DNA kit. The reliability of each DNA extraction was evaluated by performing three independent experiments using eight replicates. The quality and quantity of the L. monocytogenes DNA were tested using both a fluorescence-based DNA measuring protocol and a specific quantitative RTi-PCR assay. The total DNA yield was similar for both extraction protocols. However, the Wizard-based protocol produced a better quality of L. monocytogenes DNA, as it enhanced the RTi-PCR results (approximately one order of magnitude greater than the Chelex-based protocol). Thus, the protocol based on the Wizard DNA kit was more suitable for L. monocytogenes DNA extraction from meat samples when sensitive and accurate results are needed.

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Volume14, Issue4

December 2006

Pages 395-404

ISOLATION OF LISTERIA MONOCYTOGENES DNA FROM MEAT PRODUCTS FOR QUANTITATIVE DETECTION BY REAL-TIME PCR

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ISOLATION OF LISTERIA MONOCYTOGENES DNA FROM MEAT PRODUCTS FOR QUANTITATIVE DETECTION BY REAL-TIME PCR

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