SCREENING METHODS: Gradient plates

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Gradient plates

A method similar to the agar dilution assay is the gradient plate technique. In this method, melted agar is dispensed into a Petri dish with one edge elevated. The agar is allowed to solidify and form a wedge. A second, overlay wedge which contains the antimicrobial is then poured. The result is a plate which contains a gradient of antimicrobial concentrations from near 0 to the maximum in the overlay. The plate may then be inoculated using a spread plate method or by parallel streaks of several strains of microorganisms. The advantage of the analysis is that the microorganism is exposed to a continuous gradient of concentrations, as opposed to the 2-fold or 10- fold dilutions used in the previous tests. Payday Loans Online

This technique was used to test the antimicrobial activity of medicinal plants, spices, and essential oils by Ting and Deibel (1992) to study the sensitivity of Listeria monocytogenes to various extracts such as clove, oregano and black pepper. A variation of the gradient plate technique utilizes a spiral plating system. This assay uses the spiral plate to deposit a radial gradient of antimicrobial on an agar plate. Because the volume of liquid deposited on any section of the plate is known, the concentration of antimicrobial may then be determined as well. A bacterial suspension is inoculated onto the surface of the plate to produce a radial streak. The MIC of the compound against the microorganism is determined by a zone of clearence on the streak.

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