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PCR detection of Listeria monocytogenes in 'gravad' rainbow trout.

https://arctichealth.org/en/permalink/ahliterature75590
Source
Int J Food Microbiol. 1997 Apr 15;35(3):281-5
Publication Type
Article
Date
Apr-15-1997
Author
H. Ericsson
P. Stålhandske
Author Affiliation
Department of Food Hygiene, Faculty of Veterinary Medicine, Swedish University of Agricultural Sciences, Uppsala, Sweden. Henrik.Ericsson@lmhyg.slu.se
Source
Int J Food Microbiol. 1997 Apr 15;35(3):281-5
Date
Apr-15-1997
Language
English
Publication Type
Article
Keywords
Animals
Base Sequence
Colony Count, Microbial
DNA, Bacterial - analysis - chemistry - genetics
Food Handling - standards
Food Microbiology
Food Poisoning - etiology
Humans
Listeria monocytogenes - genetics - isolation & purification
Oncorhynchus mykiss - microbiology
Polymerase Chain Reaction - methods
Research Support, Non-U.S. Gov't
Sensitivity and specificity
Spermidine - analysis
Sweden
Time Factors
Abstract
'Gravad' rainbow trout artificially contaminated with Listeria monocytogenes was analyzed by use of a 4 h enrichment period followed by extraction of DNA and PCR amplification. This procedure made it possible to detect 10-100 cfu L. monocytogenes per gram 'gravad' rainbow trout, within 12 h. After a prolonged enrichment period of 24 h, numbers as low as 1-10 cfu L. monocytogenes per gram could be detected. The method described may be a useful tool for screening samples of 'gravad' rainbow trout for the presence of L. monocytogenes, since it is sensitive, rapid and simple.
PubMed ID
9105939 View in PubMed
Less detail

Sample preparation and DNA extraction procedures for polymerase chain reaction identification of Listeria monocytogenes in seafoods.

https://arctichealth.org/en/permalink/ahliterature11066
Source
Int J Food Microbiol. 1997 Apr 15;35(3):275-80
Publication Type
Article
Date
Apr-15-1997
Author
A. Agersborg
R. Dahl
I. Martinez
Author Affiliation
Norwegian Institute of Fisheries and Aquaculture N-9005 Tromso, Norway.
Source
Int J Food Microbiol. 1997 Apr 15;35(3):275-80
Date
Apr-15-1997
Language
English
Publication Type
Article
Keywords
Animals
Base Sequence
DNA, Bacterial - analysis - chemistry - genetics
Decapoda (Crustacea) - microbiology
Detergents - pharmacology
Electrophoresis, Agar Gel
Endopeptidase K - pharmacology
Fish Products - microbiology
Food Microbiology
Food Poisoning - diagnosis - epidemiology - etiology
Gene Amplification
Heat
Humans
Listeria monocytogenes - drug effects - genetics - isolation & purification
Muramidase - pharmacology
Norway - epidemiology
Octoxynol - pharmacology
Polymerase Chain Reaction - methods
Prevalence
Research Support, Non-U.S. Gov't
Sensitivity and specificity
Time Factors
Abstract
Five grams of seafood products were inoculated with one to 500 viable or 10(9) heat-killed cells of Listeria monocytogenes. The presence of the pathogen was detected by the polymerase chain reaction (PCR) with primers specific for fragments of the listeriolysin O (hly) gene (two sets) and for the invasion-associated protein (iap) gene (one set). For DNA preparation, boiling, either alone or in combination with lysozyme and proteinase K treatment, was not always sufficient to lyse L. monocytogenes, while treatment with Triton X-100 produced consistently good DNA suitable for amplification. To avoid false-negative and false-positive results, 48 h incubations were necessary and a subculturing step after an initial 24 h incubation greatly improved the results. The primers that amplified regions of the listeriolysin O gene gave clearer and stronger products than primers for the invasion-associated protein gene. Using this method we were able to detect one to five L. monocytogenes cells in 5 g of product in a total of 55 h.
PubMed ID
9105938 View in PubMed
Less detail