Optimized biomembrane living bacteria counting method
A technology for counting live bacteria and biofilm, which is applied in the field of optimized biofilm counting method, can solve the problems of decreased sensitivity of antibacterial agents, large differences among tested individuals, unfavorable screening work, etc., and achieves obvious dispersion effect and repeatability Strong, improve the effect of live bacteria count results
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[0091] Preparation method of dental plaque biofilm
[0092]The preparation mode of dental plaque biofilm involved in the present invention is as follows:
[0093] (1) Streak culture for each oral pathogenic bacteria on a TSA-supplemented medium plate in an oxygen-free atmosphere at a constant temperature of 36°C for 2 to 6 days, until a large number of bacterial lawns appear on the plate;
[0094] (2) Inoculate each cultured oral pathogenic bacteria into BHI-enriched medium, and culture them in an oxygen-free atmosphere at a constant temperature of 36°C for 1 to 3 days until the medium is cloudy, indicating that the culture of the bacterial suspension is mature ;
[0095] (3) Mix the mature bacterial suspension into the BHI enriched medium added with 0.2% adhesion promoter according to the volume ratio of 2% to 8%, and inoculate the TC in an oxygen-free atmosphere at a constant temperature of 36°C. In the small wells of the treated cell culture plate (24-well plate), a den...
Embodiment 1
[0218] Specifically include the following steps:
[0219] 1) Culture of dental plaque biofilm
[0220] According to the steps in the preparation example of the dental plaque biofilm model, the dental plaque biofilm was cultivated in the TC-treated cell culture plate. Calcium carbonate suspension was added as an adhesion promoter, and the incubation time was 16 hours.
[0221] 2) Antibacterial testing of oral care products using dental plaque biofilm
[0222] Draw TC to handle the bacterial suspension in the two small holes of the cell culture plate, and discard it; add 0.2mL~1mL oral care product samples of a certain concentration along the hole wall (the test sample solution in this embodiment is the test sample solution of the toothpaste sample) , keep for 3 minutes, draw the sample, and discard it; immediately add an equal volume of BHI-enriched medium to the small well along the wall of the hole, absorb the BHI-enriched medium, and discard it; then add it twice along the...
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