Simultaneous Enrichment of Salmonella spp, Escherichia coli O157:H7, Vibrio parahaemolyticus, Staphylococcus aureus, Bacillus cereus, and Listeria monocytogenes by Single Broth and Screening of the Pathogens by Multiplex Real-time PCR

被引：21

作者：

Kobayashi, Hiroshi ^{[1
]}

Kubota, Jun ^{[1
]}

Fujihara, Kumiko ^{[1
]}

Honjoh, Ken-ichi ^{[1
]}

Ito, Masayoshi ^{[1
]}

Fujiki, Naomi ^{[2
]}

Nakabe, Mika ^{[2
]}

Oda, Shun-ichi ^{[2
]}

Satoyama, Toshiya ^{[2
]}

Takasu, Kazushige ^{[3
]}

Nakanishi, Hisao ^{[3
]}

Miyamoto, Takahisa ^{[1
]}

机构：

[1] Kyushu Univ, Lab Food Hyg Chem, Div Food Biotechnol, Dept Biosci & Biotechnol,Fac Agr,Higashi Ku, Fukuoka 8128581, Japan

[2] Japan Food Res Labs, Kyushu Branch, Hakata Ku, Fukuoka 8120034, Japan

[3] Japan Food Res Labs, Osaka Branch, Suita, Osaka 5640051, Japan

来源：

FOOD SCIENCE AND TECHNOLOGY RESEARCH | 2009年 / 15卷 / 04期

关键词：

simultaneous screening; multiplex real-time PCR; single-enrichment; food poisoning; UNIVERSAL PREENRICHMENT BROTH; POLYMERASE-CHAIN-REACTION; ASSAY; O157-H7; GROWTH; FOODS; SPP; IDENTIFICATION; AMPLIFICATION; YERSINIA;

D O I：

10.3136/fstr.15.427

中图分类号：

TS2 [食品工业];

学科分类号：

0832 ;

摘要：

Simultaneous enrichment broth (SEB) was developed for the single-enrichment simultaneous screening of six, major food poisoning bacteria. After enrichment in SEB for 18 h at 37 degrees C, viable counts of six major food poisoning bacteria (Staphylococcus aureus, Salmonella spp., Escherichia coli O157:H7, Vibrio parahaemolyticus, Bacillus cereus, and Listeria monocytogenes) were sufficient for 5' nuclease multiplex real-time PCR assay using existing primers and probes. By labeling the probes with three different fluorescent dyes, the assay could be carried out in 2 tubes. The whole process, including enrichment and PCR, was completed within 24 h and the detection limits for the target bacteria from the food sample (boiled chicken) were 36 cfu/25 g for S. aureus, 5.3 cfu/25 g for Salmonella spp, 2.9 cfu/25 g for E. coli O157:H7, 2.0 cfu/25 g for V. parahaemolyticus, 5.5 cfu/25 g for B. cereus, and 6.2 cfu/25 g for L. monocytogenes. The results were comparable to conventional methods that require 4-6 days.

引用

页码：427 / 438

页数：12