山羊莱氏无胆甾原体FJ-NP株的分离鉴定

Isolation and Identification of Acholeplasma laidlawii FJ-NP Strains in Goat

为明确福建省某羊场发生的疑似羊支原体性肺炎病例的病原,利用支原体培养基对发病羊肺脏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,并对分离株16SrRNA进行测序分析。结果显示,分离株菌落呈油煎蛋状,有棕黄色中心突起;能发酵葡萄糖,不能水解精氨酸,不分解尿素,氯化四氮唑还原反应、胆固醇需要试验、血细胞吸附试验及溶血试验的结果均为阴性,美兰还原反应阳性。经PCR扩增出莱氏无胆甾原体支原体(Acholeplasma laidlawii,AL)大小为505bp的特异性目的片段,分离株16SrRNA序列与莱氏无胆甾原体标准株PG8同源性为99.8%。鉴定结果表明,本次从山羊肺脏组织分离到的支原体为莱氏无胆甾原体,命名为FJ-NP株,但该分离株与山羊发病性的关系有待进一步研究。

To confirm the pathogen of a suspected mycoplasmal pneumonia of goat in a farm of Fu-jian province,the pathogen in the lung tissue was isolated and purified using medium for Myco-plasma .It was identified by biochemical test and PCR method,and 1 6S rRNA gene of the isolate was sequenced.The results showed that the isolate colonies were like fried eggs with brown pro-trusion in the center,and it could ferment glucose,not hydrolyze arginine and decompose urea, meanwhile,four azole nitrogen chloride reduction reaction,cholesterol test,hemadsorption test, blood adsorption experiment and hemolysis test of isolate were all negative,however,Meilan re-duction reaction was positive.The productions of PCR amplification was about 505 bp which was Acholeplasma laidlawii specific band.The result of sequence analysis indicated that there was 99.8% homology between the nucleotide sequence of 1 6S rRNA gene of the isolate and that of Acholeplasma laidlawii strain PG8.The identification results showed that the isolated from the lung tissue of goat was Acholeplasma laidlawii called FJ-NP strain,while futher study would be needed for the relationship between the pathogentic isolates and goat disease.