猪肺炎支原体山西株的分离与鉴定

Isolation and Identification of Shanxi Strain of Mycoplasma hyopneumoniae

研究旨在获得猪肺炎支原体山西地方株,以期补充猪肺炎支原体山西地方株的菌株信息,为山西猪支原体肺炎的诊断及防治提供理论依据。将具有猪支原体肺炎典型病理特征的肺脏剪成米粒大小,放入KM2液体培养基,盲传培养后在固体培养基中纯化培养,用16S rRNA和种特异性基因P36进行PCR检测,对P36扩增结果测序与登录NCBI GenBank的猪肺炎支原体进行同源性比较,并构建系统发育树。结果显示,液体培养基变色提示培养基中含有猪肺炎支原体;菌落中间暗、边缘光滑,呈典型的"煎蛋样",大小0.2μm左右,生理生化特性符合支原体特性;16S r RNA和P36基因经PCR扩增确认分离菌株为猪肺炎支原体,并命名为JZ01株。JZ01株是从山西最近发病且致病性严重的猪肺中分离而来,其不仅能成功适应人工培养基,而且传代生长良好,为潜在的疫苗菌株。

The purpose of this research is to obtain the local strain of Mycoplasma hyopneumoniae in Shanxi,to supplement the strain information of the local strain of Mycoplasma hyopneumoniae in Shanxi,and to provide a basis for the diagnosis and prevention of Mycoplasma hyopneumoniae of swine in Shanxi.The lungs with typical pathological features of Mycoplasma hyopneumoniae of swine were cut to the size of rice grains and placed in KM2 liquid medium for blind transmission culture and purified and cultured in solid medium.16 S rRNA and species-specific gene P36 were used for PCR detection,and the phylogenetic tree was constructed by comparing the sequence of P36 amplification results with the homology comparison of Mycoplasma hyopneumoniae registered in NCBI GenBank.The results showed that the liquid medium discoloration indicated that the medium contained Mycoplasma hyopneumoniae.The dark edge of the colony was smooth and it was a typical"fried egg-like"with a size of about 0.2μm.The physiological and biochemical characteristics were consistent with the characteristics of Mycoplasma.The PCR amplification of 16 S rRNA and P36 genes confirmed that the isolated strain was Mycoplasma hyopneumoniae and named it JZ01 strain.The strain JZ01 was isolated from the pig lungs with recent onset and serious pathogenicity in Shanxi.It not only successfully adapts to the artificial culture medium,but also grows well in subculture,and it is a potential vaccine strain.