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Rapid Sample Processing for Detection of Food-Borne Pathogens via Cross-Flow Microfiltration

机译:通过交叉流微滤快速检测食品中致病菌的样品

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This paper reports an approach to enable rapid concentration and recovery of bacterial cells from aqueous chicken homogenates as a preanalytical step of detection. This approach includes biochemical pretreatment and prefiltration of food samples and development of an automated cell concentration instrument based on cross-flow microfiltration. A polysulfone hollow-fiber membrane module having a nominal pore size of 0.2 μm constitutes the core of the cell concentration instrument. The aqueous chicken homogenate samples were circulated within the cross-flow system achieving 500- to 1,000-fold concentration of inoculated Salmonella enterica serovar Enteritidis and naturally occurring microbiota with 70% recovery of viable cells as determined by plate counting and quantitative PCR (qPCR) within 35 to 45 min. These steps enabled 10 CFU/ml microorganisms in chicken homogenates or 10~(2) CFU/g chicken to be quantified. Cleaning and sterilizing the instrument and membrane module by stepwise hydraulic and chemical cleaning (sodium hydroxide and ethanol) enabled reuse of the membrane 15 times before replacement. This approach begins to address the critical need for the food industry for detecting food pathogens within 6 h or less.
机译:本文报道了一种方法,该方法能够从鸡匀浆中快速浓缩和回收细菌细胞,作为检测的分析前步骤。这种方法包括食品样品的生化预处理和预过滤,以及基于错流微滤的自动细胞浓缩仪的开发。标称孔径为0.2μm的聚砜中空纤维膜组件构成了细胞浓缩仪的核心。将鸡匀浆含水样品在错流系统中循环,以达到500到1,000倍的接种肠炎沙门氏菌血清肠炎沙门氏菌和天然菌群的浓度,通过平板计数和定量PCR(qPCR)确定活菌的回收率达到70% 35至45分钟。这些步骤使鸡匀浆中的10 CFU / ml微生物或10〜(2)CFU / g鸡得以定量。通过逐步进行水力和化学清洁(氢氧化钠和乙醇)清洁和消毒仪器和膜组件,可以在更换之前将膜重复使用15次。这种方法开始满足食品工业在6小时或更短时间内检测到食品病原体的关键需求。

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