Preparation of test bacteria sheet for moisture barrier microbial penetration tester

The moisture barrier microbial penetration tester is suitable for evaluating the performance of materials in resisting the penetration of bacteria in liquids when subjected to mechanical friction. It is mainly used for medical surgical drapes, surgical gowns and clean clothes. Applicable standards: YY/T 0506.6, ENISO 22610 Main parameters: 1. Turntable speed: 60rpm±1rpm; 2. Test pressure: 3N±0.02N; 3. Outward wheel speed: 5～6rpm; 4. Timer setting range: 0～99.99min; 5. Total weight of inner and outer rings: 800g±1g. Plate preparation: Staphylococcus aureus ATCC29213 on tryptic soy agar at 36°C±Incubate at 1°C for 18-24h, inoculate 2-3 colonies into 3ml of tryptic soy broth, and inoculate at 36°C±Incubate at 1°C for 18-24h. Dilute 1:10 with peptone water to a concentration of 1×104CFU/mL~4×104CFU/mL, count the final bacterial suspension. Open the sterile bag and take out the polyurethane film still attached to the IQ, and place the wettable polyurethane film of the bacteria-carrying material sheet face up on the cleanliness plate. For ease of operation, cover the four corners of the bacteria-carrying material sheet with double-sided tape fixed on the plate. Use the petri dish cover as a template to form a corresponding area on the biofilm, apply 1.0 ml of Staphylococcus aureus suspension to this area, and then dry the bacterial sheet at 56°C for about 30min, using sterile glass during the drying period. The applicator continues to coat the bacterial suspension on the bacterial carrier film to make the bacterial liquid evenly distributed. Test principle: Put the test piece on the agar petri dish, place a piece of bacteria of the same size on the test piece, and then cover it with a piece of high-density polyethylene film with a thickness of about 10 microns, and use a tapered steel ring to seal the three-layer material ( The test material is at the bottom, the bacterial sheet is in the center, and the high-density polyethylene film is on the top), and the ring kit is placed on the agar petri dish on the turntable. The test finger acts on the material in a way that can move on the entire surface of the petri dish. , subjecting the material to the combined action of pressure and friction to simulate the possible stress and microbial penetration of the barrier material in the actual application process. The microorganisms on the bacterial sheet penetrate the test material and migrate to the surface of the agar medium. The penetration performance of the test material can be quantitatively evaluated by culturing the agar plate and counting the colonies.
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