辽宁绒山羊副结核分枝杆菌的分离鉴定及分子生物学检测

Isolation,Identification and Molecular Biological Detection of Mycobacterium Tuberculosis from Liaoning Cashmere Goats

为了对某辽宁绒山羊饲养场疑似副结核病例作出正确诊断,本研究针对疑似病羊的内脏组织和肠内容物在Dubods油酸琼脂培养基上经过37℃条件下9～13周培养,从其肝脏组织中分离获得1株细菌,对该分离菌株开展了形态学观察、抗酸染色镜检、PCR检测及基因序列测定等研究工作。结果显示,分离菌为抗酸染色阳性菌,而且形态学符合副结核分支杆菌的特征; PCR检测结果为阳性,测定的序列与Gen Bank中JⅡ-1961(CP022105. 1)基因序列同源性高达100%,证实了该分离菌株为副结核分支杆菌。

In order to make a correct diagnosis of suspected tuberculosis cases in one Liaoning cashmere goat farm, we conducted the study on the Dubods oleic acid agar medium for 9-13 weeks at 37 ℃ for the visceral tissue and intestinal contents of suspected sheep, A strain was isolated from its liver tissue. Morphological observation, acid-resistant staining, PCR detection and gene sequen- cing were carried out. The results showed that the isolates were positive for acid-fast staining and the morphology was consistent with the characteristics of Mycobacterium tuberculosis. The results of PCR were positive and the sequence was 100% homologous to J II - 1961 （CP022105. 1 ） gene sequence in GenBank, Confirming that the isolate was Mycobacterium tuberculosis.