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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。
Objective To study the immunological characteristics of recombinant Mycobacterium tuberculosis protein 38KDa (rTPA38) in order to find a specific diagnostic agent. Methods Skin test wheel method: respectively HRV whole cell, BCG, Kansas, sporadic, Mycobacterium tuberculosis sensitized guinea pig spare; 5IU international 37 standard PPD as a positive control, 0.2μg recombinant rTPA38 antigen were in the dorsal skin 48h injection observed swelling, induration of the vertical and horizontal diameter. Using rTPA as antigen and PPD as control, the specific anti-TB antibodies in serum were detected by ELISA. Results Both rTPA and PPD induced late-type hypersensitivity (DTH) in dolphin-3838 mice, which produced similar DTH responses as PPD but did not produce DTH response to guinea pigs infected with Kansas, sporadic or Mycobacterium bovis. 238 cases of pulmonary tuberculosis group, rTPA and PPD detection sensitivity were 65.1%, 72.7%. The serum levels of rTPA and rTPA were 95.6%, 95.9% and 383888.5% respectively in healthy blood donors, non-tuberculosis 38 respiratory disease patients and BCG vaccinated patients. The specificity of PPD was 91.8% and 85.4 %, 68.7%. The statistical results showed that the positive rate of rTPA protein and PPD in non-tuberculosis respiratory disease group was significantly different (P <0.05). There was a significant difference between the positive rate of BCG vaccination and the serum antibody titer (P <0.05). Conclusion The DTH response of rTPA38-induced guinea pigs is similar to that of PPD. The rTPA protein antigen has high specificity and sensitivity. It is a reliable antigen of 38 ELISA, which can replace PPD as a new specific diagnostic preparation.