中文摘要：为解决现有技术中缺乏高效分离和培养猪肺炎支原体的培养基，相关支原体分离困难等技术问题，本发明旨在提供一种猪肺炎支原体的固体培养基及其制备方法，以及探究了其在分离猪肺炎支原体中的应用。本发明的猪肺炎支原体培养基能够有效地从猪肺灌洗液中分离和鉴定猪肺炎支原体，其相比传统的猪肺炎支原体的分离和鉴定方法具有更高的准确性。表明本发明的固体培养基能够有效地用于猪肺炎支原体的分离和鉴定。
外文摘要：Solid medium comprises basal medium and auxiliary medium. The basal medium is 25-35 ml 20xHank's, 2-3 g PPLO broth powder, 2-5 g yeast extract, 0.5-3 g whey *protein* hydrolysate, 3-5 g Brain heart infusion powder, 0.5-1 ml 1% zinc chloride solution, 18-22 g agar powder, 10 ml 0.25% phenol red solution and 850-900 ml deionized *water*. The auxiliary medium is 3-4 ml 5% arginine solution, 2-3 ml 5% cysteine solution, 1-2 ml 1% taurine solution, 1-5 ml 5% coenzyme I, 5-10 ml 100000 units/ml penicillin, and 70-90 ml inactivated healthy pig serum.   INDEPENDENT CLAIMS are also included for: (1) preparing the solid medium comprises (i) adding 20x Hank's, PPLO broth powder, yeast extract, whey *protein* hydrolysate, brain heart infusion powder, 1% zinc chloride solution, 0.25% phenol red solution , agar powder and deionized *water*, stirring evenly, adjusting the pH value to 7.2-7.4 with 10M sodium hydroxide, autoclaving for 15 minutes at 120 degrees C, (ii) filtering and sterilizing 5% arginine solution, 5% cysteine solution, 1% taurine solution, 5% coenzyme, 100,000 U/ml penicillin, and 56 degrees C Co-60 irradiated healthy pig serum, and (iii) cooling when the basal medium to 50-55 degrees C, adding auxiliary, stirring uniformly, and pouring into a 90 mm dish, and each plate is 10-15 ml, and cooling; and (2)#separating *Mycoplasma* hyopneumoniae, adopting solid *culture* medium for separating *culture* of *Mycoplasma* hyopneumoniae, the pig lung tissue lavage or lung tissue homogenate with 0.22 mu *m* filter to filter, uniformly coating the filtrate on *Mycoplasma* hyopneumoniae solid *culture* medium of the invention, placing at 37 degrees C *cell* *culture* box, carbon dioxide concentration of 5% to *culture* for 2-4 days, observing its shape by *electron* *microscope* *mycoplasma* will exhibits a fried egg morphology on solid *culture* medium after inoculating for 2-4 days.    Preferred Components: The solid medium comprises basal medium and an auxiliary medium. The basal medium is 30 ml 20xHank's, 2.5 g PPLO broth powder, 3 g Yeast extract, 2 g Whey *protein* hydrolysate, 4 g Brain heart infusion powder, 0.75 ml 1% zinc chloride solution, 20 g agar powder, 10 ml 0.25% phenol red solution and 870 ml deionized *water*. The auxiliary medium is 3.5 ml 5% arginine solution, 2.5 ml 5% cysteine solution, 1 ml 1% taurine solution, 3 ml 5% coenzyme I, 8 ml 100,000 U/ml penicillin, and 75 ml inactivated healthy pig serum.    The solid medium of *Mycoplasma* hyopneumoniae is useful for *isolating* *Mycoplasma* hyopneumoniae for *vaccine* preparation.
申请号：CN201810768901.3
公开/公告号：CN108949623A
申请日：2018-07-13
公开/公告日：2018-12-07
申请/专利权人：北京农学院
发明/设计人：张永红;崔德凤;薛振华;沈红
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