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Response surface methodology to design a selective co-enrichment broth of Escherichia coli, Salmonella spp. and Staphylococcus aureus for simultaneous detection by multiplex PCR

文献类型： 外文期刊

作者： Zhang, Qiao Yan 1 ; Zhou, Wen Wu 2 ; Zhou, Yu 1 ; Wang, Xiao Fu 1 ; Xu, Jun Feng 1 ;

作者机构： 1.Zhejiang Acad Agr Sci, Inst Qual & Stand Agroprod, Hangzhou 310021, Zhejiang, Peoples R China

2.Sinous E China Pharmaceut Co, Fermentat Engn Lab, Hangzhou 310011, Zhejiang, Peoples R China

关键词： Response surface methodology;Selective co-enrichment;Multiplex PCR;Food-borne pathogen;Inhibitor

期刊名称：MICROBIOLOGICAL RESEARCH （ 影响因子：5.415； 五年影响因子：6.038 ）

ISSN： 0944-5013

年卷期： 2012 年 167 卷 7 期

页码：

收录情况： SCI

摘要： Escherichia coli, Salmonella spp. and Staphylococcus aureus are frequent co-visitors of contaminated foods to cause food-borne diseases. To achieve rapid detection of three organisms by multiplex PCR, a selective co-enrichment broth was considered to design using response surface methodology (RSM) in this work. NaCl, LiCl and KSCN as selective bacterial inhibitors were selected to optimize their concentrations for a matched composition of bacterial biomass with uniform amplification of three targets. Central composite design was employed to collect the data and fit the responses. Three quadratic polynomial models were derived by computer simulation. A statistical analysis was carried out to explore the effects of the variables on the composition of bacterial biomass and PCR amplification yields. In the end, a novel broth (ESS-3 broth) of NaCl 1.60%, LiCl 0.70%, KSCN 0.10% was formulated to allow co-enrichment of the target pathogens and suppress growth of some non-target pathogens. The simultaneous detection of E. coli, Salmonella spp. and S. aureus was developed on a rapid, convenient and sensitive method consisting of selective co-enrichment in ESS-3 broth, DNA extraction with the boiling method and robust test by multiplex PCR. Our work provided broader application of RSM for the simultaneous detection of other combinations of multiple pathogens. (C) 2012 Elsevier GmbH. All rights reserved.

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