Simultaneous Screening of 24 Target Genes of Foodborne Pathogens in 35 Foodborne Outbreaks Using Multiplex Real-Time SYBR Green PCR Analysis-Reference-Cited by-同舟云学术

Simultaneous Screening of 24 Target Genes of Foodborne Pathogens in 35 Foodborne Outbreaks Using Multiplex Real-Time SYBR Green PCR Analysis

Published:2010 Issue: Volume:2010 Page:1-18
ISSN:1687-918X
Container-title:International Journal of Microbiology
language:en
Short-container-title:International Journal of Microbiology

Author:

Fukushima Hiroshi¹,Kawase Jun¹,Etoh Yoshiki²,Sugama Kumiko³,Yashiro Shunshuke⁴,Iida Natsuko⁵,Yamaguchi Keiji⁶

Affiliation:

1. Shimane Prefectural Institute of Public Health and Environmental Science, 582 Nishihamasada, Matsue, Shimane 690-0122, Japan

2. Fukuoka Institute of Health and Environmental Sciences, 39 Mukaizano, Dazaifu, Fukuoka 818-0135, Japan

3. Fukushima Institute of Public Health, 16-6 Houkida-aza-mitouchi, Fukushima 960-8560, Japan

4. Kumamoto Prefectural Institute of Health and Environmental Science, 1240-1 Kurisaki, Udo, Kumamoto 869-0425, Japan

5. Shizuoka Institute of Environment and Hygiene, 4-27-2 Kitayasuhigashi, Aoi, Shizuoka 420-8637, Japan

6. Hokkaido Institute of Public Health, West 12, Nortb 19, North Ward, Sapporo, Hokkaido 060-0819, Japan

Abstract

A set of 8 multiplex real-time SYBR Green PCR (SG-PCR) assays including 3 target primers and an internal amplification control (IAC) primer was simultaneously evaluated in 3 h or less with regard to detection of 24 target genes of 23 foodborne pathogens in 7 stool specimens of foodborne outbreak using a 96-well reaction plate. This assay, combined with DNA extraction (QIAamp DNA Stool Mini kit), offered detection of greater than103-104foodborne pathogens per g in stool specimens. The products formed were identified using melting point temperature (Tm) curve analysis. This assay was evaluated for the detection of foodborne pathogens in 33 out of 35 cases of foodborne outbreak, using 4 different PCR instruments in 5 different laboratories. No interference from the multiplex real-time SG-PCR assay, including IAC, was observed in stool specimens in any analysis. We found multiplex real-time SG-PCR assay for simultaneous detection of 24 target genes of foodborne pathogens to be comprehensive, rapid, inexpensive, accurate, of high selectivity, and good for screening probability.

Funder

Ministry of Health, Labour and Welfare

Publisher

Hindawi Limited

Subject

Microbiology (medical),Microbiology

Link

http://downloads.hindawi.com/journals/ijmicro/2010/864817.pdf

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