摘要
目的用本实验室所建立的高效液相色谱分枝菌酸分析方法,构建18种分枝杆菌标准菌株的分枝菌酸指纹谱库,并对该库的分枝菌酸指纹谱进行比较。方法标准菌株按常规方法,分别转种到改良罗氏培养基斜面上培养,待菌落形成后,用化学法从培养物中提取细胞壁的分枝菌酸,经皂化、酸化和衍生后,制备出良好的分析样品,用于色谱检测分析。结果成功地构建了18种分枝杆菌标准菌株的分枝菌酸指纹谱库,该指纹谱的重复性日内变异系数为0．20％-0．83％,均<1％。每种分枝菌酸的指纹谱都具有各自的特征,峰形也有一定规律。根据峰形的特点将其分为3类:A类(单组峰)包括结核、牛、戈登、爱知、堪萨斯、海分枝杆菌和卡介苗,B类(2组峰)包括蟾蜍、土地、新金色、不产色和浅黄分枝杆菌,C类(3组峰)包括胞内、鸟、金色、偶然、副偶然、脓肿和耻垢分枝杆菌。结论依据18种分枝杆菌标准菌株的分枝菌酸指纹谱,即可对分枝杆菌菌种进行鉴定。卡介苗与牛分枝杆菌的分枝菌酸指纹谱的峰形存在很大差别,该现象很可能作为结核病潜伏感染和分枝杆菌耐药机制的一种新理论依据。
Objective A clinical chromatographic fingerprinting of 18 standard strains of Mycobacteria commercialized in China by high-performance liquid chromatography approach was developed and improved in this laboratory. A comparison of clinical chromatographic fingerprintings of mycolic acids was carried out. Methods Mycolic acids were extracted from whole cultured cells of Mycobacteria by biochemical methods, and then samples was prepared for analysis after the mycolic acids were saponified, acidified and derived. The samples were detected by high-performance liquid chromatography under optimum conditions. Results Using above techniques a clinical chromatographic fingerprinting of Mycolic acids analysis was successfully established. The repeatability of the mycolic acids fingerprinting was evaluated. Coefficient of variation （CV） of chromatographic fingerprinting was between 0. 20% -0. 83%. Conclusions The first clinical chromatographic fingerprinting of 18 standard strains of Mycobacteria in China was established in this laboratory. Mycobacteria can be accurately identified according to chromatographic fingerprinting in a short time. Moreover, there was significant difference between BCG vaccine and Mycobacterium bevis when their peak was compared, which may lead to research of new tools for identifying insidious infection of tuberculosis and for the study of the mechanism of drug tolerance.
出处
《中华结核和呼吸杂志》
CAS
CSCD
北大核心
2006年第7期470-473,共4页
Chinese Journal of Tuberculosis and Respiratory Diseases
基金
天津市卫生局重点基金资助项目(99KY012)
关键词
色谱法
高压液相
分枝杆菌属
分枝菌酸
Chromatography, high pressure liquid
Mycobacterium
Mycolic acids
