Requirements for sterile rooms
The sterile room is generally a small room specially set up in the microbiology laboratory, the area should not be too large, about 4m2~5m2, and the height is about 2.5m. A buffer room, a clean air shower room and a dressing room should be set up outside the aseptic room. Both the sterile room and the buffer room must be airtight. Indoor equipment must have air filtering devices. Both the sterile room and the buffer room are equipped with ultraviolet lamps, and the ultraviolet lamp in the sterile room is lm away from the work surface. The floor and walls of the aseptic room must be flat, not easy to hide dirt, and easy to clean. The surface of the workbench must be level. Workers entering the sterile room should wear sterilized clothes, hats, and shoes in the dressing room, pass through the clean air shower, then to the buffer room, and then enter the sterile room for inspection work.
Enterprises in county-level cities generally use small ultra-clean benches in their laboratories. The ultra-clean workbench has a simple structure and is easy to move. There are ultraviolet lamps and a purification fan inside the workbench. The outside is a small push-pull door or a glass door with two holes. You can extend your arms in during operation.
Generally, there are the following requirements for sterile rooms:
(1) The cleanliness of the aseptic operation room should reach class 10,000, the indoor temperature should be kept at 20-24℃, the humidity should be kept at 45%~60%, and the cleanliness of the ultra-clean bench should reach class 100. The number of colonies should be checked monthly in the sterile room. When the sterile room or ultra-clean workbench is open, take a number of sterile petri dishes with an inner diameter of 90mm, and inject the plate count agar that hzas melted and cooled to about 45°C aseptically. Put 15mL of the culture medium until it solidifies, then place it upside down in a 36℃±1℃ incubator for 48 hours. After the sterility is proved, take 3 to 5 plates and place them in the left, middle and right places of the working position. Open the lid and expose for 30 minutes. Minutes later, place it upside down in an incubator at 36°C ± 1°C for 48 hours, then take it out for inspection. The average number of bacteria on the plate in the 100-level clean area shall not exceed 1 colony, and the average in the 10,000-level clean room shall not exceed 3 colonies. If it exceeds the limit, the sterile room shall be thoroughly disinfected until the repeated inspection meets the requirements.
(2) The sterile room should be kept clean and tidy. Only necessary inspection equipment such as alcohol lamp, alcohol cotton, lighter, tweezers, inoculation loop, scissors and glass pens should be stored in the room. It is strictly forbidden to pile up debris to prevent pollution.
(3) The sterile room should be equipped with a disinfectant of working concentration, such as 5% cresol solution, 75% alcohol, 0.1% neocerizine solution and so on.
(4) All items such as instruments, medicines, glassware and so on that need to be brought into the sterile room for use should be tightly wrapped or sterilized by appropriate methods. For example, the petri dishes and straws are simply packed in stainless steel and sterilized in a 160℃-170℃ dry box for 1h~2h; the physiological saline is sterilized at 121℃ for 20 minutes with an autoclave, etc.
(5) Before using the sterile room, the UV lamp in the sterile room and the buffer room must be irradiated for more than 30 minutes, and the purification fan must be turned on for ventilation at the same time. Only half an hour after the UV lamp is turned off can you enter the sterile room for work. After the sample inspection is completed, the remaining samples, various medicines and glassware should be cleaned up in time, and after the safe exit, all the UV lamps should be turned on and sterilized by irradiation for 30 minutes.
(6) The outer packaging of the samples submitted for inspection shall be kept intact before the inspection, and shall not be opened to prevent contamination. Before the test, disinfect the outer surface with 75% alcohol cotton ball, if necessary, use ignited alcohol cotton to sterilize.
(7) During each operation, a blank control should be done to check the reliability of aseptic operation.
(8) When sucking the bacteria liquid, it must be sucked by the ear ball. Do not touch the straw directly with the mouth. If the straw touches the hand or other non-sterilized objects, it must be replaced to prevent contamination.
(9) Before and after each use, the inoculation needle must be sterilized by flame combustion, and the culture can be inoculated after cooling down.
(10) Suction straws, test tubes and other utensils with bacterial liquid should be immersed in a sterile bucket containing 5% Lysol solution for disinfection, and then taken out and rinsed within 24 hours; petri dishes need to be re-sterilized in an autoclave. When handling the nutrient base, remember not to flush it directly into the sewer to prevent clogging. If any bacterial liquid is spilled on the table or on the ground, immediately use 5% carbolic acid solution or 3% Lysol to overturn on the contaminated area for at least 30 minutes, and then do the treatment. When working clothes and hats are contaminated by bacterial liquid, they should immediately Take off and wash after autoclaving.