Antimicrobial Testing of Dry Surfaces through Large Droplet Inoculation

Protocol Exchange
Lukasz PorosaDaniel Foucher

Abstract

The Large Droplet Inoculation (LDI) protocol for testing antimicrobial coatings and treatments is simple, reproducible, and closely mimics real-world conditions at solid/air interfaces. This is an advancement of the current ISO 22196/JIS Z 2801 standard method, as it provides greater ease-of-use and more closely resembles the contamination of common surfaces. The protocol involves inoculating 100 µL droplets containing ~107 microbial cells of a test microorganism onto sets of antimicrobial-treated and untreated control sample material. The sets are protected from dust accumulation and allowed to air dry, which causes all cells within each droplet to encounter the sample surface. The surviving cells are then collected by vortexing in a collection liquid and enumerating to evaluate differences in survival on the antimicrobial-treated and untreated control samples. Overall, the LDI protocol requires 10 hours of basic microbiology over one week and provides a simple means of assessing the efficacy of various antimicrobial coatings and treatments.

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