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A Verification Method for Autoclave

In addition to a small number of medium only heating and dissolving, without autoclaving, most of the medium are required 121 ℃ autoclave 15-30 minutes. Especially sterile test medium sterilization is not complete, directly related to the test results. Therefore, it is necessary to carefully verify the sterilization effect of the autoclave.

1. experiment material

(1) Bacillus stearothermophilus. Bacillus stearothermophilus ATCC7053 is a biological indicator bacteria commonly used in this method and contains 5 to 10 CFU / tablet of buds.


121 ℃ pressure steam sterilization chemical indicator card.

(3) bromocresol peptone water medium 116 ℃ autoclaved 20 minutes after the standby.

(4) O-150 ℃ leave point thermometer.

2. Methods and results

Bacillus stearothermophilus tablets (hereinafter referred to as bacteria tablets) with sterile tweezers into the sealed tube. Chemical indicator card and stay thermometer into the open test tube. The above two kinds of test tubes prepared 5-10 copies. Respectively, placed in the autoclave steam mouth, the bottom of the exhaust and the bottom of the outlet or up and down about 5 in the middle. If the sterilizer for the two, you need to put 10.

Leave the temperature gauge after passing the test can be used to verify the test. Before testing, the mercury column of the thermometer should be thrown to below 40 ℃. The temperature difference between the thermostat should be stored at 1 ° C after each monitoring, indicating that the temperature distribution within the sterilizer is uniform.

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After sterilization of the bacteria should be strictly sterile operating conditions into the sterilized bromocresol peptone water culture medium 56-60 ℃ culture 24-48 hours, observe the color change. If the medium becomes yellow, indicating that the bacteria in the Bacillus stearothermophilus has not been completely inactivated, the bacteria can still grow in the medium, decomposition of glucose acid into yellow. If the color of the culture medium does not change is still purple, then the sprouts have been inactivated. At the same time, the untreated paper was used as the positive control and the blank medium without paper was used as the negative control.

Chemical indicator card on the indicator block, in the high-pressure steam sterilization, from light yellow to black. With the color changes in the depth, and compared with the control color, can determine whether the sterilization effect to meet the requirements. Chemical instructions should be stored in a dry place. The tide will change color, affecting the observation of the sterilization effect.

High-pressure steam sterilization must make the steam smoothly into the sterilizer, contact with the sterilized items, and the original cold air can be achieved to achieve the effect of sterilization. To carry out no-load heat distribution and full heat penetration test (full load does not exceed the total volume of 2/3). Two kinds of verification of repeated three times, a total of six times.

5 points six tests showed that the temperature is 121 ℃, chemical instructions card black; degree consistent with the control color; medium did not change the color, indicating high pressure steam sterilization qualified.

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