从湖羊肺脏中分离绵羊肺炎支原体的鉴定

从新疆湖羊肺脏病料中分离出一株支原体XJ-3f，用其培养物人工感染80日龄健康绵羊，28d后剖杀，剖检可见肺表面肉粉色实变，显微病理变化为大灶性融合性肺炎。参考国际已知支原体16SrRNA序列，设计一对扩增700bp片段的通用引物，直接提取肺脏组织DNA进行PCR扩增并克隆、测序。将该序列与GenBank中33种支原体序列比较，结果证明该序列与绵羊肺炎支原体（M．ovipneumonia）标准株Y．98同源性为99．9％，而与山羊支原体山羊亚种（M．capricolum subsp．capricolum，Mcc）、丝状支原体山羊亚种（M．mycoides subsp．Capri，Mmc）、丝状支原体丝状亚种LC型（M．mycoides subsp．mycoides LC，M．mmLC）等同源率为81％。以感染羊血清和Y．98与从发病羊肺脏中分离出的三株支原体茵体蛋白进行Western blot，证明均与感染羊血清有特异性反应，且与Y．98相比无明显差异，故确定分离株为绵羊肺炎支原体（M．ovipneumonia）。

Identification of a strain of Mycoplasma ovipneumonia from the lung of a neumonia sheep

One Mycoplasma isolate XJ-3f was isolated from the lung of diseased sheep with the respiratory symptoms. The strain was cultured in modified Hayrick Medium and the culture were infected intratracheal to 2 sheep of 80 days. Lobular catarrhal bronchopneumonia was established by postmortem examinations, 28 d from infection, and Mycoplasma was re-isolated from the lungs. We designed a pair of primers to amplify a 700 bp fragment of 16S rRNA gene of all Mycoplasma. The genomic DNA of lung were extracted, amplified by the primers and cloned into the pMD18-T vector. 33 reference strains were loaded down from GenBank, the result of sequence analysis indicated that the isolated strain, XJ-3f, exhibited 99.9 % homology with reference strain Y-98, 81% with reference strains Mccp, MmmLC, Mcc ,Mmc. The result of Westem blot showed that they are all recognized by the serum as seem as Y-98. The results indicated that the isolated strain XJ-3f is belong to the Mycoplasma ovipneumonia.