猴源性结核分枝杆菌的分离和鉴定

目的探讨非人灵长类结核病的诊断方法，以及进行结核病的病原学的调查研究。方法利用病理学诊断方法，微生物分离方法以及分子生物学技术对结核菌素阳性的4只恒河猴，1只食蟹猴进行疾病诊断和病原分离。结果5只猴尸检，出现典型的干酪样病变；病变组织切片，显微镜检，可见典型的结核病特征。利用米氏7H9培养基和改良罗氏培养基进行病原分离，通过TCH和PNB等培养基进行生化鉴定，确认所分离的分枝杆菌是结核分枝杆菌；巢式PCR对石蜡包埋的病变组织抽提的DNA进行扩增，5只猴的DNA样品都扩增出结核分枝杆菌特异的123bp片段，证实感染猴的病变组织中含有结核分枝杆菌。结论非人灵长类死后，结核病的诊断可以通过常规的病理和微生物分离诊断方法进行。分子生物学技术在结核病的诊断中的应用逐渐成为必不可少的诊断方法，它将有利于对病理学诊断、病原分离困难的小的病变组织的诊断，以及对病原分离过程中分枝杆菌污染的判定。

Isolation and identification of Mycobacterium tuberculosis from monkeys

To find out suitable diagnostic method for the investigation on the etiological study of non-human primate tuberculosis, etiological diagnosis and isolation of the causative agents was performed in 4 Rhesus monkeys and a Cynomolgus monkey that was proved to be positive in the intradermal tuberculin testing ,by pathological, microbiological and molecular biological techniques.In these 5 monkeys, typical caseous lesions could be found on autopsy, and the typical characteristics of tuberculosis were demonstrated under microscopical examinations. Tubercle bacilli was isolated and identified by cultivation with Middlebrook 7H9 and modified Loewenstein'media and biochemical identification with TCH and PNB media. By using the nest-PCR amplified a region of 123 bp from the insertion sequence IS6110,M.tuberculosis was detected in the formalin-fixedparaffin-embedded sections of these monkeys. It is evident that the molecular biological techniques are required in the etiological diagnosis of tuberculosis in monkeys in addition to the previously used pathological and microbiological methods.