Mainly used in this experiment the rapid culture of Mycoplasma pneumoniae, the use of standard strains of Mycoplasma pneumoniae, Mycoplasma pneumoniae preparing the basis of the growth medium, and then conducted through the medium of the way single-factor experiments to determine culture conducive to the growth of Mycoplasma pneumoniae needed nutrients, anti-miscellaneous growth factor and the environment, and then selected the optimal growth of Mycoplasma pneumoniae results showed that: the growth of Mycoplasma pneumoniae cultured 3-5 days, medium yellow color, has begun to demonstrate mycoplasma pneumoniae specific phenomenon of the normal growth; miscellaneous experiments showed that access to anti-bacteria (such as the type of people support the original body, Ureaplasma urealyticum, Neisseria gonorrhoeae, fungi, etc.), the medium color process control bacteria did not change; of the negative control, the medium has always been the process of change without any color. Experimental results show that: fetal calf serum, yeast extract, glucose and other components contribute to the growth of Mycoplasma pneumoniae culture; penicillin, ammonia benzyl vancomycin, amphotericin, such as antibiotics can be effective in inhibiting bacteria growth medium, there are good the ability of anti-miscellaneous.