Virtual colony count (VCC) is a kinetic, 96-well microbiological assay originally developed to measure the activity of defensins.[1] It has since been applied to other antimicrobial peptides including LL-37.[2] It utilizes a method of enumerating bacteria called quantitative growth kinetics, which compares the time taken for a bacterial batch culture to reach a threshold optical density with that of a series of calibration curves. The name VCC has also been used to describe the application of quantitative growth kinetics to enumerate bacteria in cell culture infection models.[3]
Antimicrobial susceptibility testing (AST) can be done on 96-well plates by diluting the antimicrobial agent at varying concentrations in broth inoculated with bacteria and measuring the minimum inhibitory concentration that results in no growth. However, these methods cannot be used to study some membrane-active antimicrobial peptides, which are inhibited by the broth itself. The virtual colony count procedure takes advantage of this fact by first exposing bacterial cells to the active antimicrobial agent in a low-salt buffer for two hours, then simultaneously inhibiting antimicrobial activity and inducing exponential growth by adding broth. The growth kinetics of surviving cells can then be monitored using a temperature-controlled plate reader. The time taken for each growth curve to reach a threshold change in optical density is then converted into virtual survival values, which serve as a measure of antimicrobial activity.
^Ericksen B, Wu Z, Lu W, Lehrer RI (2005). "Antibacterial Activity and Specificity of the Six Human α-Defensins". Antimicrob. Agents Chemother. 49 (1): 269–75. doi:10.1128/AAC.49.1.269-275.2005. PMC 538877. PMID 15616305.
^Pazgier M, Ericksen B, Ling M, Toth EA, Shi J, Li X, Galliher-Beckley A, Lan L, Zou G, Zhan C, Yuan W, Pozharski E, Lu W (2013). "Structural and functional analysis of the pro-domain of human cathelicidin, LL-37". Biochemistry. 52 (9): 1547–58. doi:10.1021/bi301008r. PMC 3634326. PMID 23406372.
^Hoffmann S, Walter S, Blume AK, Fuchs S, Schmidt C, Scholz A, Gerlach RG (2018). "High-Throughput Quantification of Bacterial-Cell Interactions Using Virtual Colony Counts". Front Cell Infect Microbiol. 8 (43): 43. doi:10.3389/fcimb.2018.00043. PMC 5818393. PMID 29497603.
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