2007 Fiscal Year Final Research Report Summary
Contamination of foods and sources of the emerging diarrheagenic Fec ericbia mallet foreshadowspandemics
| Project/Area Number |
18602001
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
食の安全
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| Research Institution | Osaka City University |
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Principal Investigator |
NISHIKAWA Yoshikazu Osaka City University, Graduate School of Human Life Science, Professor (60183539)
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| Co-Investigator(Kenkyū-buntansha) |
OGASAWARA Jun Osaka City Institute of Public Health and Environmental Sciences, Department of Microbiology, Senior Researcher (00270760)
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| Project Period (FY) |
2006 – 2007
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| Keywords | darrheagenic / Escherichia coli / real-time PCR / multiplex / food / screening / epidemiology / contamination |
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| Research Abstract |
Diarrheagenic Escherichia coli (DEC) comprise a variety of pathotypes: enteropathogenic E. coli (EPEC); shigatoxin-producing E. coil (STEC); enterotoxigenic E. coli (ETEC); enteroinvasive E. coli (EIEC); enteroaggregative E. call (EAggEC); and diffusely adhering E. coil (DAEC). However, the source and routes of infection have not been clarified because these organisms are scarcely detected among numerous coliform bacteria. To exhaustively detect DEC in food, we have developed multiplex real-time PCR assays against enterovirulent genes (eae, stx1, stx2, elt, est, virB, aggR, astA and afaB). Primers and TaqMan probes were designed to amplify and quantify the genes in duplex or triplex reactions under the same PCR cycling conditions. Specificity was confirmed using 96 DEC strains, 2 strains of Vibrio cholerae El Tor Ogawa, and Shigella dysenteriae serotype 1. The fluorescence threshold cycle and DNA concentrations correlated with decision coefficients of more than 0.99. Subsequently, ground meat samples and enrichment broths were spiked with DEC. Detection limits varied from 3.6×10^2 to 6.0×10^3 CFU/ml, depending on the target genes. All meat samples spiked with a variety of DEC (more than 10 CFU/10g) were found to be positive by screening the enrichment broth using multiplex real-time PCR. As a preliminary test, 148 food samples were exhaustively assayed by multiplex real-time PCR; 41 samples (28%) were positive for DEC pathotypes. The present multiplex real-time PCR system allows the efficient and simultaneous determination of various DEC pathotypes in food.
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[Journal Article] Enhancement of Shiga toxin production in enterohemorrhagic Escherichia coli serotype O157:H7 by DNase colicins2007
Author(s)
Toshima, H., Yoshimura, A., Arikawa., K., Hidaka, A., Ogasawara, J., Hale, A., Masaki, H., and Nishikawa, Y.
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Journal Title
Appl.Environ.Microbiol. 73
Pages: 7582-7588
Description
「研究成果報告書概要(和文)」より
Peer Reviewed
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[Journal Article] Association of IL-8-Inducing strains of diffusely adherent Escherichia coli with sporadic diarrheal patients less than 5 years of age2007
Author(s)
Meraz, I.M., Arikawa, K., Nakamura, H., Ogasawara, J., Hase, A., the Surveillance Team, and Nishikawa, Y.
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Journal Title
Braz.J.Infect.Dis. 11
Pages: 44-49
Description
「研究成果報告書概要(和文)」より
Peer Reviewed
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[Journal Article] Prevalence of enteric bacteria that inhibit growth of enterohaemorrhagic Escherichia coli 0157 in humans2007
Author(s)
Toshima, H., Y., Hachio, M., Ikemoto, Y., Ogasawara, J., Hase, A., Takahashi, K., Masaki, N., and Nishikawa, Y.
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Journal Title
Epidemiol. Infect. 135
Pages: 110-117
Description
「研究成果報告書概要(和文)」より
Peer Reviewed
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[Journal Article] Enhancement of Shiga toxin production in enterohemorrhagic Escherichia coil serotype 0157:H7 by DNase colicins2007
Author(s)
Toshima, H, Yoshimura, A, Arikawa, K, Hidaka, A, Ogasawara, J, Hase, A, Masaki, H, Nishikawa, Y
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Journal Title
Appl. Environ. Microbiol 73
Pages: 7582-7588
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Association of IL-8-Inducing strains of diffusely adherent Escherichia coli with sporadic diarrheal patients less than 5 years of age2007
Author(s)
Meraz, I. M, Arikawa, K, Nakamura, H, Ogasawara, J, Hase, A, the Surveillance, Team, Nishikawa, Y
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Journal Title
Braz. J. Infect. Dis. 11
Pages: 44-49
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Prevalence of enteric bacteria that inhibit growth of enterohaemorrhagic Escherichia coil ○ 157 in humans2007
Author(s)
Toshima, H., Y. Hachio, M., Ikemoto, Y. Ogasawara, J. Hase, A. Takahashi, K. Masaki, H. and Nishikawa, Y
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Journal Title
Epidemiol. Infect. 135
Pages: 110-117
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Investigation of diarrhoeic faecal samples for enterotoxigenic,Shiga toxin-producing and typical or atypical enteropathogenic Escherichia coli in Kashmir,India2006
Author(s)
Wani, S.A., Nabi, A., Fayaz, I., Ahmad, I., Nishikawa, Y., Qureshi, K., Khan, M.A., and Chowdhary, J.
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Journal Title
FEMS Microbiol.Lett. 261
Pages: 238-244
Description
「研究成果報告書概要(和文)」より
Peer Reviewed
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[Journal Article] Investigation of diarrhoeic faecal samples for enterotoxigenic, Shiga toxin-producing and typical or atypical enteropathogenic Escherichia coli in Kashmir, India2006
Author(s)
Wani, S. A, Nabi, A, Fayaz, I, Ahmad, I. Nishikawa, Y, Qureshi, K, Khan, M. A, Chowdhary, J
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Journal Title
FEMS Microbiol. Lett. 261
Pages: 238-244
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Multiplex real-time PCR for exhaustive detection of diarrheagenic Escherichia coil in foods2007
Author(s)
Hidaka, A, Hokyo, T, Ogasawara, J, Hase, A, Takatori, K, Nishikawa, Y
Organizer
The 28th Annual Meeting of Japanese Society of Food Microbiology
Place of Presentation
Tokyo
Year and Date
20070926-27
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Simultaneous detection of diarrheagenic Escherichia coli(EHEC, EPEC,ETEC)in foods by real-time PCR2006
Author(s)
Hidaka, A, Hokyo, T, Ogasawara, J, Hase, A, Nishikawa, Y
Organizer
The 27th Annual Meeting of Japanese Society of Food Microbiology
Place of Presentation
Osaka
Year and Date
20060921-22
Description
「研究成果報告書概要(欧文)」より
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