Laboratory of Renewable Resources Engineering.
Appl Environ Microbiol. 2013 Nov;79(22):7048-54. doi: 10.1128/AEM.02587-13. Epub 2013 Sep 6.
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.
本文报道了一种方法,可作为检测的分析前步骤,从水相鸡匀浆中快速浓缩和回收细菌细胞。该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化细胞浓缩仪器的开发。标称孔径为 0.2 μm 的聚砜中空纤维膜组件构成了细胞浓缩仪器的核心。水相鸡匀浆样品在错流系统中循环,实现了接种的肠炎沙门氏菌血清型肠炎和自然存在的微生物菌群的 500-1000 倍浓缩,通过平板计数和定量 PCR(qPCR)在 35 至 45 分钟内可回收 70%的活菌。这些步骤使鸡匀浆中 10 CFU/ml 的微生物或鸡中 10(2) CFU/g 的微生物能够被定量。通过逐步水力和化学清洗(氢氧化钠和乙醇)清洗仪器和膜组件,并对膜进行消毒,可在更换之前重复使用 15 次。该方法开始满足食品行业在 6 小时或更短时间内检测食源性病原体的迫切需求。