Multiplex real-time PCR detection of heat-labile and heat-stable toxin genes in enterotoxigenic Escherichia coli

被引:17
作者
Grant, MA
Hu, JX
Jinneman, KC
机构
[1] US FDA, Pacif Reg Lab NW, Bothell, WA 98021 USA
[2] US FDA, Seafood Prod Res Ctr, Bothell, WA 98021 USA
[3] Washington State Publ Hlth Lab, Shoreline, WA 98155 USA
关键词
D O I
10.4315/0362-028X-69.2.412
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
A multiplex real-time PCR method was developed for detection of heat-labile and heat-stable toxin genes in enterotoxigenic Escherichia coli. Approximately 10 CFU per reaction mixture could be detected in rinsates from produce samples. Several foods representative of varieties previously shown to have caused enterotoxigenic E. coli outbreaks were spiked and enriched for 4 or 6 h. Both heat-labile and heat-stable toxin genes could be detected in the foods tested, with the exception of hot sauce, with threshold cycle values ranging from 25.2 to 41.1. A procedure using membrane filtration which would allow enumeration of the enterotoxigenic E. coli population in a food sample in less than 28 h by real-time PCR analysis of colonies picked from media highly selective for E. coli was also developed.
引用
收藏
页码:412 / 416
页数:5
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