Isolation and culture identification of Mycoplasma
Isolation and culture identification of Mycoplasma
Mycoplasma isolation and culture identification is used to (1) detect different biochemical reactions of mycoplasmas ter (2) isolate and identify mycoplasmas.
Operation method
Isolation and culture identification of Mycoplasma
Principle
The specimens are inoculated separately within the Mycoplasma identification culture. Mycoplasma urealyticum produces urease, which leads to the production of ammonia from sugar, the culture medium becomes alkaline, and the indicator changes from yellow to pink. Mycoplasma hominis can break down arginine to produce nitrogen, which makes the culture medium containing arginine broth alkaline and pink.
Materials and Instruments
Bacterial samples Move 【Specimen collection】Specimens should be sent for examination immediately after collection, and stored at room temperature for no more than 2 hours, and at 2-8°C for no more than 5 hours. Male: insert a male sterile swab into the urethral opening at 2-4cm to gently rotate the secretion; prostate fluid or semen culture can also be taken. If necessary, urine can also be used for mycoplasma culture, take 10-20 ml of the middle part of the first urine in the early morning, centrifuged at 2000 rpm for 10 minutes, and take the sediment for inoculation, but the positive rate of urine mycoplasma culture is significantly lower than that of urethral secretion. Women: after dilating the cervix with a dilator, use a female swab to gently rotate the cervical secretion 1-2cm inside the cervical opening; amniotic fluid can also be used for culture. Urine is not recommended. Culture 1, culture tube rewarming take out the required number of culture tubes, rewarm to room temperature, make a good mark. 2, inoculation will be swab inserted into the culture tube, rotate and squeeze several times, so that the swab in the mycoplasma infiltrated into the liquid, the swab is sterilized and properly disposed of; urine sediment, semen, amniotic fluid, prostate fluid and other liquid specimens take 0.2 ml of inoculation culture solution, mix thoroughly. 3, culture 35-37 ℃ constant temperature culture 24-48h hours, observe the results. Caveat 1, Uu, Mh growth should keep the culture fluid transparent and clear, no obvious turbidity, occasional turbidity shall not be reported Mycoplasma positive. 2， Uu growth, the normal color change time of the culture medium in 24 hours, but if the specimen content is small, the color change time may be more than 24 hours.Mh growth, the normal color change time of the culture medium in 24-48 hours, if the specimen content is large, the color change time may be less than 24 hours. 3， can not determine whether there are stray bacteria contamination, can be taken out of the culture solution a little inoculation in the blood plate or through the 0.45um filter membrane filtration culture, according to the growth of blood plate colonies or filtered after the culture of the color change to determine the stray bacteria contamination. Common Problems Result determination: Uu should be reported within 24 hours, Mh should be reported within 48 hours, and other results should be reported within 48 hours, with a maximum of 72 hours. Culture tube liquid from orange to red, transparent and clear, without obvious turbidity, indicating that there is Uu and/or Mh growth, can be judged as positive; culture tube liquid without any change, can be reported as negative results; culture liquid obviously turbid and red, should be judged no mycoplasma growth. If necessary, solid culture of mycoplasma can be done, and the growth of mycoplasma can be observed under low-power microscope as mycoplasma-positive if there is egg-like colony growth. For more product details, please visit Aladdin Scientific website.
Mycoplasma isolation and identification medium
Thermostat Incubator Culture Tubes Centrifuge