Optimization of an amplified system for the detection of <i>Clostridium tyrobutyricum</i> on nitrocellulose filters by use of monoclonal antibody in a gelified medium

The normal procedure for bacterial colony detection requires a nitrocellulose transfer step after membrane filtration and culture to prevent the development of a high background during the immunodetection. In this paper, we describe a modification of the basic protocol that omits the transfer step and reduces the risk of background. Previous observations indicated that interactions between milk components (principally cream) and membrane are responsible for the high non-specific staining observed. Experiments were performed to remove lipid components or to block the membrane binding sites before milk filtration. Samples of milks of different origin (collected at different times of the year) and different membranes were tested. The results obtained showed that removing lipids did not significantly improve the test but, on the contrary, led to an antigen diffusion. Incubation of the membrane in 0.1% (w/v) of Tween 20 in phosphate-buffered saline before milk filtration prevented non-specific binding, and allowed performance of the detection without any noticeable background.

show abstract

Optimization of an amplified system for the detection of Clostridium tyrobutyricum on nitrocellulose filters by use of monoclonal antibody in a gelified medium

Cited by 7 publications

References 14 publications

CLOSTRIDIUM | Clostridium tyrobutyricum

CLOSTRIDIUM | Clostridium tyrobutyricum

Relevance and analysis of butyric acid producing clostridia in milk and cheese

Monoclonal antibody detection of Clostridium microcolonies directly on membrane used for milk filtration

Contact Info

Product

Resources

About