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Rapid detection of bacterial cells using photon correlation spectroscope technique on HACCP

HACCP 中利用光子相关光谱技术快速检测细菌细胞

基本信息

批准号：
13660201
负责人：
ENDO Hideaki
金额：
$ 2.5万
依托单位：
Tokyo University of Marine Science and Technology
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

项目摘要

We developed new system for rapid detection of bacteria cells using photon correlation spectroscope (PCS) technique.1.Removal of bacterial cells by ultrasonic vibrationTo remove bacterial cells from the surface of seafood products (Surimi-based products, fish), ultrasonic energy was used. Almost of all cells can be removed from seafood in 5 min with ultrasonic treatment. Flow cytometry (FCM) technique applied to the determination of total number of microbial cells grown on seafood products. Cell numbers determined by FCM well paralleled that measured by traditional colony counting method in the range of 105-109 cells/g. One FCM assay could be completed within 60 sec and the total assay time including the preparation of microbial cells was within 30 min.2.Rapid detection of Salmonella cells by PCS techniqueRapid detection system for nitrifying bacteria was developed using PCS technique. The method is based on that Salmonella cells reacted with anti-Salmonella antibody were able to analyze by PCS technique. When Salmonella cells were reacted with the antibody beads, the size distribution was shifted to larger range. In contrast, in the reference bacterial cells (Escherichia coli), the size distribution shows little change. The phenomenon was assumed that since the antibody reacted with Salmonella cells selectively, they were aggregated with bacterial cells, and the distribution size increased. Thus, it is possible to discriminate between aimed nitrifying bacteria and other spices of bacterial cells using PCS technique. One assay could be completed in 10 min, and the total PCS assay time including sample preparation was within 60 min.

我们开发了一种新的光子相关光谱（PCS）技术快速检测细菌细胞的系统。1.超声波振动去除细菌细胞为了从海产品（基于鱼糜的产品，鱼）表面去除细菌细胞，使用超声波能量。超声波处理可以在5 min内去除几乎所有的细胞。应用流式细胞仪（FCM）技术测定海产品上微生物细胞总数。流式细胞仪检测细胞数，传统菌落计数法检测细胞数在105-109个/g之间。一次FCM检测可在60 s内完成，包括细胞制备在内的总检测时间在30 min内。2.应用PCS技术快速检测沙门氏菌该方法是基于沙门氏菌细胞与抗沙门氏菌抗体反应，能够通过PCS技术进行分析。当沙门氏菌细胞与抗体珠反应时，尺寸分布向更大的范围移动。相比之下，在参考细菌细胞（大肠杆菌）中，大小分布几乎没有变化。该现象被认为是由于抗体选择性地与沙门氏菌细胞反应，它们与细菌细胞聚集，并且分布尺寸增加。因此，利用PCS技术可以区分目标硝化细菌和其他种类的细菌细胞。一次测定可在10 min内完成，包括样品制备在内的总PCS测定时间在60 min内。