丝状支原体山羊亚种FJ-GT株的分离和鉴定

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，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，用16S rRNA通用引物对分离株进行克隆测序。将分离菌株命名为FJ-GT。结果显示菌落呈典型的油煎蛋状，中心有棕褐色突起；分离株能发酵葡萄糖，不能水解精氨酸，不分解尿素，血细胞吸附试验呈阴性，胆固醇需要试验、美兰还原反应和氯化四氮唑还原反应均呈阳性，溶血试验为β溶血；PCR结果扩增出丝状支原体山羊亚种(Mycoplasma mycoidessubsp. capri, Mmc)特异大小为394 bp 的目的片段；菌株16S rRNA 序列与丝状支原体山羊亚种标准株PG3 的序列比较，同源性为99.5%。鉴定结果表明本次分离到的支原体为丝状支原体山羊亚种。本研究首次从病原学角度证明了福建省存在由丝状支原体山羊亚种引起的羊支原体性肺炎，对福建省羊支原体性肺炎的诊断和有效防控具有重要的指导意义。

Isolation and Identification of Mycoplasma mycoides subsp. capri FJ-GT Strain

To confirm the pathogen of a suspected contagious caprine pleuropneumonia case in a goat farm of Fujian Province, the pathogen of lung tissue was isolated and purified using mycoplasma medium. It was identified by biochemical test and specificity polymerase chain reaction (PCR). Cloning and sequencing studies were given to the sequence of the isolated 16S rRNA. The results showed that bacterial colonies were like fried eggs with brown protuberance in the center. The isolated strain was named FJ-GT and could ferment glucose, but could not hydrolyze arginine and decompose urea. Hemadsorption test of FJ-GT was negative. Meilan reduction reaction, the cholesterol test and four azole nitrogen chloride reduction reaction of FJ-GT were positive. Hemolysis test of FJ- GT showed a beta hemolysis. The result of the PCR amplification of Mycoplasma mycoides subsp. capri was the specific band of 394 bp. Sequence analysis indicated that 16S rRNA of the isolated FJ-GT shared 99.5% homology with Mycoplasma mycoides subsp. capri strain PG3. Thus it could be concluded that the pathogen was Mycoplasma mycoides subsp. capri. This study proved theexistence of goat mycoplasmal pneumonia caused by Mycoplasma mycoides subsp. capri from the perspective of etiology in Fujian Province for the first time, and had guiding effect on the diagnosis, prevention and control of mycoplasmal pneumonia of sheep and goats in Fujian Province.