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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Research Article|
August 01 2004
Optimization of Rapid Detection of Escherichia coli O157:H7 and Listeria monocytogenes by PCR and Application to Field Test
GI-SEONG MOON;
GI-SEONG MOON
Food Safety Team, Korea Food Research Institute, Seongnam, Kyunggi 463-746, Republic of Korea
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WANG JUNE KIM;
WANG JUNE KIM
Food Safety Team, Korea Food Research Institute, Seongnam, Kyunggi 463-746, Republic of Korea
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WEON-SUN SHIN
WEON-SUN SHIN
*
Food Safety Team, Korea Food Research Institute, Seongnam, Kyunggi 463-746, Republic of Korea
* Author for correspondence. Tel: +82-(0)31-780-9125; Fax: +82-(0)31-709-9876; E-mail: hime@kfri.re.kr.
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J Food Prot (2004) 67 (8): 1634–1640.
Article history
Received:
July 30 2003
Accepted:
February 03 2004
Citation
GI-SEONG MOON, WANG JUNE KIM, WEON-SUN SHIN; Optimization of Rapid Detection of Escherichia coli O157:H7 and Listeria monocytogenes by PCR and Application to Field Test. J Food Prot 1 August 2004; 67 (8): 1634–1640. doi: https://doi.org/10.4315/0362-028X-67.8.1634
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