Isolation and Identification of Mycoplasma ovipneumoniae FJ01-CL
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摘要: 为明确福建省某羊场发生的疑似山羊传染性胸膜肺炎病例的病原,利用支原体培养基对肺组织的病原进行分离培养和纯化,通过生化试验和特异性PCR方法进行鉴定,结果显示菌落呈“桑葚状,无中心脐”;分离株能发酵葡萄糖,不能水解精氨酸,不分解尿素,美兰还原反应阳性,氯化四氮唑还原反应阳性,血细胞吸附试验阳性,胆固醇需要试验阳性;PCR结果扩增出绵羊肺炎支原体特异大小为361bp的目的片段,将该序列同GenBank中8株绵羊肺炎支原体序列进行比较,结果证明该序列同绵羊肺炎支原体标准株Y-98同源性为98.1%,与地方株MoGH3-3的同源性最高,为98.7%。鉴定结果表明本次分离到的支原体为绵羊肺炎支原体,并命名为FJ01-CL株。本研究首次证明了福建省存在由绵羊肺炎支原体(Mycopalsam ovipneumonia,Mo)引起的羊支原体性肺炎。Abstract: To confirm a suspected contagious pleuropneumonia case at a sheep farm in Fujian,the pathogen from the diseased sheep lung tissue was isolated and purified on a mycoplasma medium.It was subsequently identified by using a biochemical test and specificity polymerase chain reaction(PCR).There were " mulberries,no center umbilical" colonies on the petri dishes observed in the laboratory.The isolated FJ01-CL could ferment glucose,but could not hydrolyze arginine,nor decompose urea.It was positive on the Meilan reduction reaction,the 4-azolenitrogen-chloride reduction reaction,the blood adsorption test and the cholesterol test.The PCR amplification of the sheep mycoplasma pneumoniae showed a specific band of 361 bp.Compared with 8sequence strains from the Genbank,the sequence analysis indicated that the isolated FJ01-CL exhibited 98.1% homology with the reference strain,Y-98,and 98.7% with the local strains,MoGH3-3.The results suggested that Mycoplasma ovipneumoniae FJ01-CL was the pathogen that caused the caprine mycoplasmal pneumonia in the province for the first time.
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Key words:
- Mycoplasma ovipneumoniae /
- isolation /
- identification /
- Fujian
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