Development of Duplex SYBR Green Real-Time PCR for Rapid and Simultaneous Detection of 16 Specific Genes of 16 Major Foodborne Bacteria
[Introduction] In foodborne outbreaks, public health administrators must respond promptly and effectively to prevent the spread of pathogens and the recurrence of food poisoning. To do this, it is necessary to identify the causative agents of outbreaks as soon as possible. Traditional culture method...
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| Published in | JAPANESE JOURNAL OF FOOD MICROBIOLOGY Vol. 30; no. 3; pp. 160 - 164 |
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| Main Authors | , , , , , , |
| Format | Journal Article |
| Language | English Japanese |
| Published |
Japanese Society of Food Microbiology
30.09.2013
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| Subjects | |
| Online Access | Get full text |
| ISSN | 1340-8267 1882-5982 |
| DOI | 10.5803/jsfm.30.160 |
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| Abstract | [Introduction] In foodborne outbreaks, public health administrators must respond promptly and effectively to prevent the spread of pathogens and the recurrence of food poisoning. To do this, it is necessary to identify the causative agents of outbreaks as soon as possible. Traditional culture methods that have been routinely used require several days to identify foodborne bacteria. Traditional culture methods are time-consuming and laborious, since the necessary culture conditions and nutrient media differ from bacteria to bacteria. Thus, simple and reasonable methods for simultaneously detecting various food-poisoning bacterial species are preferable. Recently, specific gene sequences have been identified in various pathogenic bacteria, and reports on PCR methods targeting the gene sequences of foodborne or waterborne pathogens have been extensively studied 4,7,9). In food poisoning tests, these PCR methods are useful for screening prior to culture. For instance, Fukushima et al. developed a method for simultaneously detecting 24 specific genes of foodborne bacteria using a relatively low-cost multiplex real-time PCR system 3). |
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| AbstractList | [Introduction] In foodborne outbreaks, public health administrators must respond promptly and effectively to prevent the spread of pathogens and the recurrence of food poisoning. To do this, it is necessary to identify the causative agents of outbreaks as soon as possible. Traditional culture methods that have been routinely used require several days to identify foodborne bacteria. Traditional culture methods are time-consuming and laborious, since the necessary culture conditions and nutrient media differ from bacteria to bacteria. Thus, simple and reasonable methods for simultaneously detecting various food-poisoning bacterial species are preferable. Recently, specific gene sequences have been identified in various pathogenic bacteria, and reports on PCR methods targeting the gene sequences of foodborne or waterborne pathogens have been extensively studied 4,7,9). In food poisoning tests, these PCR methods are useful for screening prior to culture. For instance, Fukushima et al. developed a method for simultaneously detecting 24 specific genes of foodborne bacteria using a relatively low-cost multiplex real-time PCR system 3). |
| Author | IIDA, Natsuko FUKUSHIMA, Hiroshi KANDA, Takashi SUGIYAMA, Kanji HIROI, Midori YAGI, Miya MURAKAMI, Masaru |
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| Copyright | 2013 Japanese Society of Food Microbiology |
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| References | 8) Nguyen, T. V., Van, P. L., Huy, C. L., Gia, K. N. and Weintraub, A.: Detection and characterization of diarrheagenic Escherichia coli from young children in Hanoi, Vietnam. J. Clin. Microbiol., 43, 755–760 (2005). 1) Bubert, A., Riebe, J., Schnitzler, N., Schönberg, A., Goebel, W. and Schubert, P.: Isolation of catalase-negative Listeria monocytogenes strains from listeriosis patients and their rapid identification by anti-p60 antibodies and/or PCR. J. Clin. Microbiol., 35, 179–183 (1997). 3) Fukushima, H., Kawase, J., Etoh, Y., Sugama, K., Yashiro, S., Iida, N. and Yamaguchi, K.: Simultaneous screening of 24 target genes of foodborne pathogens in 35 foodborne outbreaks using multiplex real-time SYBR Green PCR analysis. Int. J. Microbiol., Article ID 864817, 17 pages (2010). 4) Fukushima, H., Tsunomori, Y. and Seki, R.: Duplex real-time SYBR Green PCR assays for detection of 17 species of food- or waterborne pathogens in stools. J. Clin. Microbiol., 41, 5134–5146 (2003). 5) Health and Welfare Department, Shizuoka Prefecture: Food poisoning in Shizuoka Prefecture 2004 (in Japanese). 2) Fukushima, H., Katsube, K., Tsunomori, Y., Kishi, R., Atsuta, J. and Akiba, Y.: Comprehensive and rapid real-time PCR analysis of 21 foodborne outbreaks. Int. J. Microbiol., Article ID 917623, 13 pages (2009). 6) Health Department, Shizuoka Prefecture: Food poisoning in Shizuoka Prefecture 2009 (in Japanese). 9) Ott, S. J., Musfeldt, M., Ullmann, U., Hampe, J. and Schreiber, S.: Quantification of intestinal bacterial populations by real-time PCR with a universal primer set and minor groove binder probes: a global approach to the enteric flora. J. Clin. Microbiol., 42, 2566–2572 (2004). 7) Matsuki, T., Watanabe, K., Fujimoto, J., Takada, T. and Tanaka, R.: Use of 16S rRNA gene-targeted group-specific primers for real-time PCR analysis of predominant bacteria in human feces. Appl. Environ. Microbiol., 70, 7220–7228 (2004). 1 2 3 4 5 6 7 8 9 |
| References_xml | – reference: 8) Nguyen, T. V., Van, P. L., Huy, C. L., Gia, K. N. and Weintraub, A.: Detection and characterization of diarrheagenic Escherichia coli from young children in Hanoi, Vietnam. J. Clin. Microbiol., 43, 755–760 (2005). – reference: 5) Health and Welfare Department, Shizuoka Prefecture: Food poisoning in Shizuoka Prefecture 2004 (in Japanese). – reference: 9) Ott, S. J., Musfeldt, M., Ullmann, U., Hampe, J. and Schreiber, S.: Quantification of intestinal bacterial populations by real-time PCR with a universal primer set and minor groove binder probes: a global approach to the enteric flora. J. Clin. Microbiol., 42, 2566–2572 (2004). – reference: 3) Fukushima, H., Kawase, J., Etoh, Y., Sugama, K., Yashiro, S., Iida, N. and Yamaguchi, K.: Simultaneous screening of 24 target genes of foodborne pathogens in 35 foodborne outbreaks using multiplex real-time SYBR Green PCR analysis. Int. J. Microbiol., Article ID 864817, 17 pages (2010). – reference: 6) Health Department, Shizuoka Prefecture: Food poisoning in Shizuoka Prefecture 2009 (in Japanese). – reference: 1) Bubert, A., Riebe, J., Schnitzler, N., Schönberg, A., Goebel, W. and Schubert, P.: Isolation of catalase-negative Listeria monocytogenes strains from listeriosis patients and their rapid identification by anti-p60 antibodies and/or PCR. J. Clin. Microbiol., 35, 179–183 (1997). – reference: 4) Fukushima, H., Tsunomori, Y. and Seki, R.: Duplex real-time SYBR Green PCR assays for detection of 17 species of food- or waterborne pathogens in stools. J. Clin. Microbiol., 41, 5134–5146 (2003). – reference: 7) Matsuki, T., Watanabe, K., Fujimoto, J., Takada, T. and Tanaka, R.: Use of 16S rRNA gene-targeted group-specific primers for real-time PCR analysis of predominant bacteria in human feces. Appl. Environ. Microbiol., 70, 7220–7228 (2004). – reference: 2) Fukushima, H., Katsube, K., Tsunomori, Y., Kishi, R., Atsuta, J. and Akiba, Y.: Comprehensive and rapid real-time PCR analysis of 21 foodborne outbreaks. Int. J. Microbiol., Article ID 917623, 13 pages (2009). – ident: 2 – ident: 3 – ident: 5 – ident: 4 – ident: 1 – ident: 6 – ident: 9 – ident: 7 – ident: 8 |
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| Title | Development of Duplex SYBR Green Real-Time PCR for Rapid and Simultaneous Detection of 16 Specific Genes of 16 Major Foodborne Bacteria |
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