Porter J, Edwards C, Morgan J A, Pickup R W
Department of Genetics and Microbiology, University of Liverpool, United Kingdom.
Appl Environ Microbiol. 1993 Oct;59(10):3327-33. doi: 10.1128/aem.59.10.3327-3333.1993.
The use of fluorescence-activated flow cytometric cell sorting to obtain highly enriched populations of viable target bacteria was investigated. Preliminary studies employed mixtures of Staphylococcus aureus and Escherichia coli. Cells of S. aureus, when mixed in different proportions with E. coli, could be selectively recovered at a purity in excess of 90%. This was possible even when S. aureus composed only approximately 0.4% of the total cells. Cell sorting was also tested for the ability to recover E. coli from natural lake water populations and sewage. The environmental samples were challenged with fluorescently labelled antibodies specific for E. coli prior to cell sorting. Final sample purities of greater than 70% were routinely achieved, as determined by CFU. Populations of E. coli released into environmental samples were recovered at greater than 90% purity. The use of flow cytometry and cell sorting to detect and recover viable target bacteria present at levels of less than 1% within an indigenous microflora was also demonstrated.
研究了使用荧光激活流式细胞仪细胞分选技术来获得高度富集的活靶细菌群体。初步研究采用了金黄色葡萄球菌和大肠杆菌的混合物。当金黄色葡萄球菌细胞与大肠杆菌以不同比例混合时，可以以超过90%的纯度选择性回收。即使金黄色葡萄球菌仅占总细胞的约0.4%，这也是可能的。还测试了细胞分选从天然湖水菌群和污水中回收大肠杆菌的能力。在细胞分选之前，用针对大肠杆菌的荧光标记抗体处理环境样品。通过CFU测定，最终样品纯度通常达到70%以上。释放到环境样品中的大肠杆菌群体以大于90%的纯度回收。还证明了使用流式细胞术和细胞分选来检测和回收本地微生物群中含量低于1%的活靶细菌。
