Optimization of rapid detection of Escherichia coli O157:H7 and Listeria monocytogenes by PCR and application to field test

Gi Seong Moon, June Kim Wang, Weon-Sun Shin

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

For rapid detection of Escherichia coli O157:H7 and Listeria monocytogenes, simple methods for sample preparation and PCR were established and applied to a field test. To improve specificity, primer sets LP43-LP44 and C(+)-D(-) were selected for E. coli O157:H7 and L. monocytogenes, respectively. Through centrifugation and partial heat treatment after enrichment, E. coli O157:H7 and L. monocytogenes were detected at 1 initial CFU without genomic DNA extraction in the culture and with artificially inoculated food samples including milk, chicken, ham, and pork. Based on the optimized PCR method, a feasibility test was carried out using randomly collected field samples. To remove false positives and false negatives, a PCR method using several primer sets, including the optimized primer set, and a standard culture method were used. With the PCR detection and standard culture methods, two pork samples were positive for L. monocytogenes after enrichment, indications that the PCR assay could be effectively used for rapid, sensitive, and species-specific detection of foodborne pathogens.

Original languageEnglish
Pages (from-to)1634-1640
Number of pages7
JournalJournal of Food Protection
Volume67
Issue number8
DOIs
StatePublished - 2004 Jan 1

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Escherichia coli O157
Listeria monocytogenes
Polymerase Chain Reaction
testing
pork
sampling
methodology
ham
food pathogens
Centrifugation
centrifugation
Chickens
Milk
foods
Hot Temperature
heat treatment
chickens
genomics
milk
Food

Cite this

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abstract = "For rapid detection of Escherichia coli O157:H7 and Listeria monocytogenes, simple methods for sample preparation and PCR were established and applied to a field test. To improve specificity, primer sets LP43-LP44 and C(+)-D(-) were selected for E. coli O157:H7 and L. monocytogenes, respectively. Through centrifugation and partial heat treatment after enrichment, E. coli O157:H7 and L. monocytogenes were detected at 1 initial CFU without genomic DNA extraction in the culture and with artificially inoculated food samples including milk, chicken, ham, and pork. Based on the optimized PCR method, a feasibility test was carried out using randomly collected field samples. To remove false positives and false negatives, a PCR method using several primer sets, including the optimized primer set, and a standard culture method were used. With the PCR detection and standard culture methods, two pork samples were positive for L. monocytogenes after enrichment, indications that the PCR assay could be effectively used for rapid, sensitive, and species-specific detection of foodborne pathogens.",
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Optimization of rapid detection of Escherichia coli O157:H7 and Listeria monocytogenes by PCR and application to field test. / Moon, Gi Seong; Wang, June Kim; Shin, Weon-Sun.

In: Journal of Food Protection, Vol. 67, No. 8, 01.01.2004, p. 1634-1640.

Research output: Contribution to journalArticle

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