摘要
目的对重组结核分枝杆菌11k Da蛋白(以下简称重组11k Da蛋白)皮肤试验与体外干扰素γ(interferonγ,IFNγ)检测方法进行比较。方法选取32名健康志愿者进行重组11k Da蛋白和TB-PPD的同体双臂皮肤试验,分别于不同前臂皮内注射0.1 ml重组11k Da蛋白(10μg/ml)和0.1 ml TB-PPD(50 IU/ml),注射后24、48、72 h测量志愿者皮肤反应(硬结和红晕)的两个直径(最大径和最大径的垂直径),并取其均值。于皮肤试验前采集32名志愿者的静脉血,分离外周血单个核细胞,采用T-SPOT.TB试剂盒进行检测。比较重组11k Da蛋白皮肤试验与体外IFNγ检测结果的一致性。结果以注射72 h后的硬结反应统计重组11k Da蛋白皮肤试验中阳性人数为8名,阳性率为25%,TB-PPD皮肤试验中阳性人数为24名,阳性率为75%,其中强阳性人数为8名;体外IFNγ检测(T-SPOT.TB)阳性人数为7名,阳性率为22%。重组11k Da蛋白皮肤试验结果和体外IFNγ检测结果一致性良好(Fleiss Kappa值=0.913),这两种检测方法的阳性率远低于TB-PPD皮肤试验(Fleiss Kappa值=0.266)。结论重组11k Da蛋白皮肤试验的检测结果与体外IFNγ检测结果基本一致,有潜力成为结核分枝杆菌潜伏感染者的新筛查方法。
Objective To compare the skin test and in vitro interferon γ（IFNγ） test with recombinant Mycobacterium tuberculosis 11 k Da protein.Methods Skin tests were performed with recombinant M.tuberculosis 11 k Da protein and TB-PPD on two arms of each of 32 volunteers respectively.Each volunteer was injected i.d.with 0.1 ml 11 k Da protein（10 μg / ml）and 0.1 ml TB-PPD（50 IU / ml） in two arms respectively,of whom the maximum diameters and their vertical diameters of duration and flush were measured 42,48 and 72 h after injection,and the means were calculated.The venous blood samples of volunteers were collected before skin test,from which peripheral blood mononuclear cells（PBMCs）were isolated and determined by T-SPOT.TB kit.The consistence of result of skin test with that of in vitro IFNγtest was evaluated.Results Positive results were observed in 8 volunteers of skin test with 11 k Da protein based on the duration reaction 72 h after injection,indicating a positive rate of 25%.However,in skin test with TB-PPD,positive results were observed in 24 volunteers,including strong positive results in 8 volunteers,indicating a positive rate of 75%.Positive results were observed in 7 volunteers of in vitro IFNγ test,indicating a positive rate of 22%.The result of skin test with recombinant 11 k Da protein was consistent with that of in vitro IFNγ test（Fleiss Kappa value = 0.913）.However,the positive rates determined by both the methods were significantly lower than that by skin test with TB-PPD（Fleiss Kappa value = 0.266）.Conclusion The skin test with recombinant M.tuberculosis 11 k Da protein showed a basically consistent result with that of in vitro IFNγtest,which might be a novel method for screening of patients potentially infected with M.tuberculosis.
出处
《中国生物制品学杂志》
CAS
CSCD
2015年第11期1183-1186,共4页
Chinese Journal of Biologicals
基金
"十二五"国家重大科技专项(2012ZX10004701)
