Objective: To study the biological characteristics of mycoplasma pneumoniae and various nutritional elements on the cultivation of Mycoplasma pneumoniae growth to explore the culture detection of Mycoplasma pneumoniae (MP) the clinical application value. Methods: Isolation and culture of the main method used, the use of standard strains of Mycoplasma pneumoniae, Mycoplasma pneumoniae preparing the basis of the growth medium, and then conducted through the medium of single factor experiment in favor of Mycoplasma pneumoniae determined by the need to foster growth nutrients, anti-miscellaneous growth factor and the environment, and then selected the optimal growth of Mycoplasma pneumoniae medium, the general environment of constant temperature 37 ℃ train 3-7 days to observe the experimental results, the selection of medium of the bacteria change the color tube, Mycoplasma pneumoniae strains in order to establish a library. Results: The positive: the growth of Mycoplasma pneumoniae cultured 3-5 days, medium yellow color, has begun to demonstrate mycoplasma pneumoniae specific phenomenon of the normal growth; anti Miscellaneous: Access bacteria (including the type of people support the original body, Ureaplasma urealyticum, Neisseria gonorrhoeae, fungi, etc.), the medium color process control bacteria did not change; negative: always culture medium without any color change, the result of normal. Conclusion: The fetal calf serum, yeast extract, glucose and other ingredients help to foster the growth of Mycoplasma pneumoniae, penicillin, ammonia benzyl vancomycin, amphotericin, such as antibiotics can inhibit effective access medium the growth of bacteria, there are good anti-Miscellaneous ability MP method can be used as the gold standard for early diagnosis, and the method is simple, rapid, easy to promote.