How to isolate medium in microbiology laboratory
1. Gram-positive bacillus isolation medium
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1. Luo-Qin modified medium
[use]
For the culture of Mycobacterium tuberculosis.
[Ascording method]
Potassium dihydrogen phosphate 2.4g, magnesium sulfate 0.24g, magnesium citrate (or sodium citrate) 0.6g, asparagine 3.6g, glycerin 12ml, distilled water 600ml, potato starch 30g, fresh chicken egg liquid 1L, 2% 20 ml of malachite green aqueous solution.
Phosphate, magnesium sulfate, magnesium citrate, aspartate and glycerin are first dissolved in 600 ml of distilled water by heating. Add the potato powder, stir it while stirring, and continue to heat it in boiling water for 30min. When it is cooled to about 60 °C, add 1L of chicken egg liquid and 20ml of malachite green solution, mix thoroughly, and then dispense with aseptically. Sterilize the test tubes, each 5~6ml, plug the rubber stopper (zui is a tipping plug), place it in a serum coagulator and make a bevel, and sterilize it intermittently at 85 °C for 1 h (or autoclave at 115 °C for 20 min). ), after sterility test after solidification, refrigerate at 4 ° C for use.
[usage]
Take morning cough or other body fluid specimens, 0.1ml (about 2 drops) of the concentrated solution after digestion and centrifugation, drop on the inclined surface of the medium, shake as much as possible, and incubate in a 5%~10% carbon dioxide incubator at 35°C. 1 to 4 weeks, observe the results. Colonies that grow in 1 week are generally unlikely to be Mycobacterium tuberculosis; colonies that grow after 2 weeks, creamy, slightly yellow, rough, opaque, that is, smear microscopy and identification .
[QC]
The M. tuberculosis strain was used as a positive culture test.
[save]
It is effective for 2~4 weeks in the refrigerator at 4 °C.
Note:
(1) This medium was modified based on the design of Lowenstein-Jenden.
(2) The pH of the medium is about 6.0, and generally no calibration is required.
(3) The temperature of intermittent sterilization should not exceed 90 °C.
2. Serum slant medium (Lu's serum bevel)
[use]
For the culture of Corynebacterium diphtheriae.
[Ascording method]
1% glucose broth (pH 7.6) 100 ml, sterile animal serum (bovine, sheep, pig, rabbit) 300 ml.
After mixing the above ingredients, they were dispensed into a test tube, and each tube was about 4 to 5 ml. Inclined in the serum coagulator (or steamer) heated 80 ~ 85 ° C for 30min, so that the serum solidified into a slope, cooled and placed in a refrigerator at 4 ° C. After taking out, it was sterilized by intermittent sterilization at 85 ° C for 30 min for 3 consecutive days. It can be applied by sterility test without growth of bacteria. 