DNA Extraction and PCR Methods for the Detection of Listeria monocytogenes in Cold-Smoked Salmon

Protocols for the specific detection of Listeria monocytogenes in cold-smoked salmon were developed. PCR was used as the method of detection. Inhibitors of PCR present in the food samples were removed by ether extraction or column purification, or their effect was overcome by the use of Tween 20 as an enhancer. These protocols are many times more rapid than conventional detection methodologies and also have the potential for automation.

[1]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[2]  K. Holmstrøm,et al.  Inhibition of PCR by components of food samples, microbial diagnostic assays and DNA-extraction solutions. , 1992, International journal of food microbiology.

[3]  S. Ratnam,et al.  Occurrence of Listeria in hot and cold smoked seafood products. , 1994, International journal of food microbiology.

[4]  M. Salimans,et al.  Rapid and simple method for purification of nucleic acids , 1990, Journal of clinical microbiology.

[5]  E. Domann,et al.  Suitability of the prfA gene, which encodes a regulator of virulence genes in Listeria monocytogenes, in the identification of pathogenic Listeria spp , 1992, Applied and environmental microbiology.

[6]  G. Stewart,et al.  Ultra sensitive detection of Listeria monocytogenes in milk by the polymerase chain reaction (PCR) , 1992, Letters in applied microbiology.

[7]  W. F. Thompson,et al.  Rapid isolation of high molecular weight plant DNA. , 1980, Nucleic acids research.

[8]  S. Ratnam,et al.  Prevalence of Listeria in Smoked Fish. , 1992, Journal of food protection.

[9]  W. Goebel,et al.  Identification of a gene that positively regulates expression of listeriolysin, the major virulence factor of listeria monocytogenes. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[10]  T. Chakraborty,et al.  Use of the polymerase chain reaction for direct detection of Listeria monocytogenes in soft cheese. , 1991, The Journal of applied bacteriology.

[11]  P. B. Ben Embarek.  Presence, detection and growth of Listeria monocytogenes in seafoods: a review. , 1994, International journal of food microbiology.

[12]  R. Buchanan,et al.  Comparison of lithium chloride-phenylethanol-moxalactam and modified Vogel Johnson agars for detection of Listeria spp. in retail-level meats, poultry, and seafood , 1989, Applied and environmental microbiology.

[13]  P. B. Embarek,et al.  Presence, detection and growth of Listeria monocytogenes in seafoods: a review. , 1994 .

[14]  R. Lacey,et al.  ACP Broadsheet 129: August 1991. Isolation and identification of Listeria monocytogenes. , 1991, Journal of clinical pathology.