评估抗体对活细菌功能亲和力的方法。

Method for assessment of functional affinity of antibodies for live bacteria.

作者信息

Brovko Lubov, Young Denise, Griffiths Mansel W

机构信息

Canadian Research Institute for Food Safety, University of Guelph, 43 McGilvray str. Guelph, Ontario, Canada N1G 2W2.

出版信息

J Microbiol Methods. 2004 Jul;58(1):49-57. doi: 10.1016/j.mimet.2004.03.002.

DOI:10.1016/j.mimet.2004.03.002

PMID:15177903

Abstract

A rapid and convenient method for assessment of functional affinity of antibodies against live bacteria is described. When a combination of immunomagnetic separation (IMS) with bioluminescent or fluorescent genetic labelling of the cells was employed, the method showed good correlation with plate count. However, the use of reporter bacteria allowed results to be obtained within 1 h compared with days using conventional methods. Due to its lower detection limit, the bioluminescent assay performed better than the fluorescent assay. Antibody affinities for Escherichia coli O157:H7 and Salmonella enteritidis were examined at different environmental conditions such as pH 3-7, temperature 4-25 degrees C, and sodium chloride concentrations 0-5% and compared with sensitivities of ELISA.

摘要

本文描述了一种快速便捷的方法，用于评估抗体对活细菌的功能亲和力。当将免疫磁珠分离（IMS）与细胞的生物发光或荧光基因标记相结合时，该方法与平板计数法具有良好的相关性。然而，使用报告细菌可在1小时内获得结果，而传统方法则需要数天。由于其检测限较低，生物发光检测比荧光检测表现更好。在不同环境条件下，如pH值3 - 7、温度4 - 25摄氏度和氯化钠浓度0 - 5%，检测了抗体对大肠杆菌O157:H7和肠炎沙门氏菌的亲和力，并与酶联免疫吸附测定（ELISA）的灵敏度进行了比较。