摘要
目的评价重组结核分枝杆菌11kDa(相对分子质量为11000)蛋白(简称"重组11kDa蛋白")在潜伏性结核感染筛查和卡介苗接种鉴别中的应用。方法由首都医科大学附属北京胸科医院联合北京市昌平区结核病防治所、北京市怀柔区结核病防治所和北京市大兴区结核病防治所,于2014年7月至2016年3月在以上3个区的高校和工厂招募健康志愿者。共计招募3001名志愿者,所有志愿者体检合格,均签订知情同意书。对所有志愿者采用Mantoux法进行重组11kDa蛋白和结核菌素纯蛋白衍生物(TB-PPD)同体双臂皮肤试验,分别于不同前臂皮内注射0.1ml重组11kDa蛋白(10μg/ml)和0.1ml TB-PPD(50U/ml),观察注射后72h后的皮肤反应。于皮肤试验前采集所有志愿者的静脉血,进行体外γ-干扰素释放试验(IGRA)检测。采用"一致率(%)"和"一阶一致性系数(the first-order agreement coefficient,AC1)比较重组11kDa蛋白皮肤试验、TB-PPD试验及IGRA结果的差异。3001名志愿者均接受3种方法检测,其中475名3种检测结果均为阴性的受试者,采用数字表法按随机双盲的原则,以2∶1的比例接种卡介苗(318名;失访8例)和安慰剂(147名;失访2例),接种3个月后再次进行重组11kDa蛋白皮肤试验、TB-PPD皮肤试验及IGRA,采用"一致率(%)"和"AC1"比较其结果差异。结果健康志愿者潜伏性结核感染筛查中,IGRA阳性率(34.4%,1033/3001)明显高于重组11kDa蛋白(32.2%,966/3001),差异有统计学意义(χ^2=22.787,P<0.001);IGRA与重组11kDa蛋白的一致率为93.4%(2804/3001),AC1值为0.882(95%CI=0.866~0.898),两者有较好的一致性。TB-PPD阳性率(47.7%,1430/3001)明显高于重组11kDa蛋白(32.2%,966/3001),差异有统计学意义(χ^2=182.146,P<0.001);TB-PPD与重组11kDa蛋白的一致率为60.6%(1819/3001),AC1值为0.243(95%CI=0.207~0.279),两者的一致性较差。卡介苗接种鉴别研究中,接种卡介苗的志愿者重组11kDa蛋白的阳性率(1.3%,4/318)与IGRA(3.1%,10/318)比较,差异无统计学意义(χ^2=2.571,P=0.109);一致率为95.6%(304/318),AC1值为0.954(95%CI=0.929~0.979),两者有较好的一致性。重组11kDa蛋白的阳性率(1.3%,4/318)低于TB-PPD(56.3%,179/318),差异有统计学意义(χ^2=167.350,P<0.001);TB-PPD与重组11kDa蛋白的一致率为42.5%(135/318),AC1值为0.025(95%CI=-0.106~0.155),两者的一致性较差。接种安慰剂的志愿者重组11kDa蛋白的阳性率(4.1%,6/147)与IGRA(1.4%,2/147)比较,差异无统计学意义(χ^2=2.667,P=0.109);一致率为95.9%(141/147),AC1值为0.957(95%CI=0.921~0.993),两者有较好的一致性。TB-PPD的阳性率(17.7%,26/147)高于重组11kDa蛋白(4.1%,6/147),差异有统计学意义(χ^2=15.385,P<0.001),一致率为82.3%(121/147),AC1值为0.781(95%CI=0.690~0.871),两者有较好的一致性。结论重组11kDa蛋白在潜伏性结核感染筛查和卡介苗接种方面与IGRA有很好的一致性,可作为潜伏性结核感染者筛查与卡介苗接种鉴别的候选诊断试剂。
Objective To evaluate the application of recombinant Mycobacterium tuberculosis 11 kDa protein(relative molecular mass 11000)in screening for latent tuberculosis infection and identification of BCG vaccination.Methods Recruited volunteers in Changping District,Huairou District,and Daxing District of Beijing from July2014 to March 2016,signed informed consent,and recruited 3001 healthy volunteers after qualified medical examination,using the Mantoux method to carry out the two-arm skin test of recombinant 11 kDa protein and TB-PPD,and intradermal injection of different forearms 0.1 ml of recombinant 11 kDa protein(10μg/ml)and0.1 ml of tuberculin pure protein derivative(TB-PPD)(50 U/ml),and observed the skin reaction 72 hours after injection.Before the skin test,venous blood of volunteers was collected for in vitro interferon gamma release assay(IGRA)detection.The"consistency rate(%)"and"the first-order agreement coefficient(AC1)"were used to compare the results of the recombinant 11 kDa protein skin test,TB-PPD test and IGRA results.Three thousand and one volunteers were tested by 3 methods,choosing 475 of subjects with negative results of all the three test,to sign an informed consent.According to the random number table double-blind principle(in the ratio of 2∶1),they were vaccinated with BCG(318 subjects)and placebo(147 subjects).After 3 months of inoculation,they were again subjected to parallel recombinant 11 kDa protein skin test,TB-PPD skin test and IGRA.Results In the screening of latent tuberculosis infection in healthy volunteers,the difference between 11 kDa(32.2%,966/3001)and IGRA(34.4%,1033/3001)results was statistically significant(χ^2=22.787,P<0.001);the consistency rate of IGRA and 11 kDa was 93.4%(2804/3001),and the Gwet’s AC1 value was 0.882(95%CI=0.866-0.898),the two had good consistency.Compared with the result of 11 kDa(32.2%,966/3001)and TB-PPD(47.7%,1430/3001),the positive rate of TB-PPD was significantly higher than that of 11 kDa.The difference between the result of TB-PPD and 11 kDa was very significant(χ^2=182.146,P<0.001).The consistency rate of TB-PPD and11 kDa was 60.6%(1819/3001),and its Gwet’s AC1 value was 0.243(95%CI=0.207-0.279).The consistency between the two was poor.In the BCG vaccination identification study,there was no statistically significant difference between the 11 kDa(1.3%,4/318)and IGRA(3.1%,10/318)results in the vaccination group(χ^2=2.571,P=0.109).The agreement rate was 95.6%(304/318),and the Gwet’s AC1 value was 0.954(95%CI=0.929-0.979),the two had good consistency.The difference between 11 kDa(1.3%,4/318)and TB-PPD(56.3%,179/318)results was very significant(χ^2=167.350,P<0.001).The consistency rate of TB-PPD and11 kDa was 42.5%(135/318),and its Gwet’s AC1 value was 0.025(95%CI=-0.106-0.155).The consistency between the two was poor.In the placebo group,there was no significant difference in the positive rate of 11 kDa(4.1%,6/147)and IGRA(1.4%,2/147)(χ^2=2.667,P=0.109);the consistency rate was 95.9%(141/147),and the AC1 value was 0.957(95%CI=0.921-0.993),which had good consistency.The positive rate of TB-PPD(17.7%,26/147)was higher than that of 11 kDa(4.1%,6/147),the difference was statistically significant(χ^2=15.385,P<0.001),the consistency rate was 82.3%(121/147),and the AC1 value was 0.781(95%CI=0.690-0.871).Conclusion Recombinant Mycobacterium tuberculosis 11 kDa protein has a good consistency with IGRA in screening for latent tuberculosis infection and BCG vaccination,suitable for large-scale screening,and can be used as one of the candidate diagnostic reagents for the screening of latent infected persons and the differentiation of BCG vaccination.
作者
赵爱华
康万里
王国治
高正伦
都伟欣
卢锦标
沈小兵
苏城
徐苗
郑素华
ZHAO Ai-hua;KANG Wan-li;WANG Guo-zhi;GAO Zheng-lun;DU Wei-xin;LU Jinbiao;SHEN Xiao-bing;SU Cheng;XU Miao;ZHENG Su-hua(Division of Tuberculosis Vaccine and Allergen,National Institutes for Food and Drug Control,Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech Products,Beijing 102629,China)
出处
《中国防痨杂志》
CAS
CSCD
2020年第8期821-825,共5页
Chinese Journal of Antituberculosis
关键词
重组蛋白质类
潜伏性结核病
试剂盒
诊断
皮肤试验
卡介苗
诊断
鉴别
Recombinant proteins
Latent tuberculosis
Reagent kits
diagnostic
Skin tests
BCG vaccine
Diagnosis
differential
