结核分支杆菌重组38KDa蛋白质抗原免疫学特性的研究

目的研究重组结核分支杆菌蛋白38KDa(rTPA38)的免疫学特性,以寻找特异性的诊断制剂。方法皮肤试验轮圈法:分别以HRV全菌体、卡介苗、堪萨斯、偶发、瘰疬分支杆菌致敏豚鼠备用;以5IU国际37标准品PPD为阳性对照,0.2μg重组rTPA38抗原分别于背部皮内注射48h观察红肿、硬结反应的纵横径。以rTPA为抗原,PPD为对照,用ELISA法检测血清中的特异性抗结核抗体。结果rTPA与PPD均可诱发豚3838鼠迟发型超敏反应(DTH),所产生的DTH反应与PPD相似,而对堪萨斯、偶发、瘰疬分支杆菌致敏的豚鼠不产生DTH反应。238例肺结核病组、rTPA和PPD检测的灵敏度分别为65.1%、72.7%。健康献血员组、非结核38呼吸疾病组和卡介苗接种阳转者血清同时用rTPA和PPD检测,rTPA的特异性分别为95.6%、95.9%、383888.5%;PPD的特异性分别为91.8%、85.4%、68.7%。统计结果表明rTPA蛋白和PPD检测非结核呼吸疾病组,其38阳性率差异有显著性(P<0.05)。两者检测卡介苗接种阳转组的阳性率和血清抗体滴度差异有显著性(P<0.05)。结论rTPA38诱发豚鼠的DTH反应与PPD相似。rTPA蛋白抗原有较高的特异性和灵敏度,是ELISA法的38可靠抗原,可替代PPD成为一种新的特异性诊断制剂。

Research in immunological characteristics of recombinant 38KDa protein of mycobacterium tuberculosis

To study the immunological characteristics of recombinant 38KDa protein (rTPA38), to find specificity of diagnostic reagent. We first inoculated strains of Mycobacterium tuberculosis(H37RV),BCG, Kansasii, Fortuitum and Scrofulaceum into guinea pig,s abdomen respectively till they got sensitive to each kind of bacteria. secondly, we took intracutaneous injection to each guinea pig with 0.2ug of rTPA38 for study, 5IU of PPD for positive control and natural saline for negative control respectively, they skin response was measured by transverse and ordinate mean diameters 48 hours after imjection. The purified rTPA38 was ured to detect sera antibodier using enzyme-linked immnosorbent assay(ELISA), and using the PPD as control. The guinea pigs delay-typed hypersensitisation (DTH) with Mycobacterium tuberculosis had a positive skin response to rTPA38 protein like PPD. The guinea pigs with Mycobacterium Kansaceii, Fortuitum, Scrofulaceum or BCG had very weak resitive to rTPA38 protein. All of these guinea pigs had positive response to PPD. In detecting the sera of 238 pulmonary tuberculosis patients, the sensitivity of rTPA38 and PPD was 65.1% and 72.7% respectively. The specificity of rTPA38 and PPD in detecting of other groups: healthy control, non-tuberculosis respivatury patients, BCG-vaccinated healthy controls, was 95.6%, 95.9%, 88.5%;and 91.8%, 85.4%, 68.7% respectively. In detecting the sera of non-tuberculosis respivatury patients, BCG-vaccinated healthy controls, rTPA38 had much more positive reactions than that of PPD. [Conclusion] rTPA38 has similar function as PPD to induce the DTH of guinea pig, rTPA38, which can provide a satisfactory sensitivity and specificity, may become effective antigen in ELISA. It may be used as a new reagent for diagnosis of tuberculosis.