Enzyme Labeled Antibody and Colony Hybridization Methods

A number of researchers have taken advantage of the orderly positioning of colonies on the HGMF to develop either research or analytical tools based on either enzyme labeled antibody (ELA) or colony hybridization. ELA-based methods developed by Todd et al. (34) and by Doyle and Schoeni (33) for detecting or enumerating E. coli 0157:H7 have already been mentioned. Peterkin et al. (51) developed a process for screening large numbers of DNA probes using HGMF. In addition, Kaysner et al. (52) reported on a DNADNA colony hybridization method to enumerate and differentiate V. parahaemolyticus and V. vulnificus, and Peterkin et al. (53) used a chromogen-labeled DNA probe to detect and confirm Listeria monocytogenes colonies on HGMF using a colony hybridization process.

Several characteristics of the HGMF contribute to its excellent performance. Principal among these are the colony-retaining and separating properties of the hydrophobic grid (23,27). The filter also serves as a passive support, enabling bacterial colonies to be transported undisturbed from one culture medium to another (32,54). Filtration often improves sensitivity of detection both by concentrating low levels of target organisms and by separating the organisms from any inhibitory materials (eg, preservatives, spice components) that might be present in the sample homogenate (27,54). The orderly grid array facilitates colony counting, thus reducing a significant source of between-analyst variation (23,27,55).

Numerous researchers have taken advantage of one or more of these HGMF characteristics to develop completely new procedures or to improve existing ones. It is inevitable that more uses will continue to be found for this highly versatile microbiological tool.

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