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区分卡介苗(BCG)和结核杆菌进而区分BCG的再激活和结核杆菌再感染是很重要的.作者将4个BCG菌株与17个结核杆菌毒力株作比较,发现25-kDa蛋白可作为BCG和结核杆菌的鉴别标志.作者先将所有的分枝杆菌接种于Lowenstein-Jensen斜面37℃培养,然后将生长的细菌从斜面上刮下并制成悬液,在沸水浴中培育15分钟后离心,最后将含可溶性蛋白的上清用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)进行分离.另外,作者用含25-kDa蛋白的溶液皮下注射瑞士白兔,21天后加强免疫.加强免疫15天后采血,获得抗25-kDa蛋白的特异性抗体.
It is important to distinguish between BCG and M. tuberculosis in order to distinguish between BCG reactivation and M. tuberculosis re-infection.The authors compared 4 BCG strains with 17 M. tuberculosis virulence strains and found that the 25-kDa protein could be used as BCG and BCG Identification of Mycobacterium tuberculosis All mycobacteria were seeded in a Lowenstein-Jensen ramp at 37 ° C and the growing bacteria were scraped off the bevel and made into suspension, which was centrifuged for 15 minutes in a boiling water bath, Finally, the soluble protein-containing supernatant was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) .In addition, the authors injected a Swiss white rabbit subcutaneously with a 25-kDa protein solution, After 15 days of booster immunization, blood was collected to obtain specific antibodies against 25-kDa protein.