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Rapid Detection of Escherichia coli O157:H7 by Loop-Mediated Isothermal Amplification Coupled with a Lateral Flow Assay Targeting the z3276 Genetic Marker

文献类型： 外文期刊

作者： Xia, Xuefeng 1 ; Zhang, Bicheng 3 ; Wang, Jing 2 ; Li, Bin 1 ; He, Kongwang 2 ; Zhang, Xuehan 1 ;

作者机构： 1.Jiangsu Univ, Sch Food & Biol Engn, 301 Xuefu Rd, Zhenjiang 212013, Jiangsu, Peoples R China

2.Jiangsu Acad Agr Sci, Inst Vet Med,Key Lab Vet Biol Engn & Technol, Minist Agr Minist,Jiangsu Key Lab Food Qual & Saf, State Key Lab Cultivat Base,Minist Sci & Technol, 50 Zhong Ling St, Nanjing 210014, Jiangsu, Peoples R China

3.Jiangsu Prov Anim Dis Prevent & Control Ctr, Nanjing 210024, Jiangsu, Peoples R China

4.Yangzhou Univ, Jiangsu Coinnovat Ctr Prevent & Control Important, Yangzhou 225000, Jiangsu, Peoples R China

关键词： E. coli O157:H7; z3276; Detection; Lateral flow assay; Loop-mediated isothermal amplification

期刊名称：FOOD ANALYTICAL METHODS （ 影响因子：3.366； 五年影响因子：3.07 ）

ISSN： 1936-9751

年卷期：

页码：

收录情况： SCI

摘要： The most commonly reported serotype of Shiga toxin-producing Escherichia coli (STEC) is O157:H7. This pathogen presents a threat to public health and is a cause of foodborne illness worldwide. The efficient and sensitive detection of E. coli O157:H7 remains a challenge for food safety. In this report, we developed a sensitive and specific loop-mediated isothermal amplification (LAMP) reaction coupled with a lateral flow (LF) assay to rapidly detect E. coli O157:H7. Six specific primers were targeted to the genetic marker z3276. Two loop primers were labeled with either fluorescein isothiocyanate (FITC) or digoxin (Dig), which facilitated analysis by the LF assay. The LAMP-LF assay detected E. coli O157:H7, and no cross-reactivity was observed with the other bacterial strains tested, including non-O157 STEC and other E. coli. The detection limit of the LAMP-LF assay was 1.3 CFU/mL for E. coli O157:H7 in broth culture, which was of equal or higher sensitivity than the other amplicon detection methods tested (agarose electrophoresis and dye). Analysis of samples from slaughterhouses indicted that the z3276-LAMP-LF assay was superior in terms of sensitivity, accuracy, and rapidity in detecting E. coli O157:H7 compared with conventional PCR. A further advantage of this method is that it is not dependent upon specialized equipment or techniques and therefore has wide potential applications in the field.

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