FadD9重组蛋白在活动性肺结核诊断中的价值  

Role of FadD9 recombinant protein for the diagnosis of active tuberculosis

作　　者：张君丽 林晨 王玉臣 刘军[1] 袁俐[2] 潘稚芬[3] 张鹭[1] ZHANG Junli;LIN Chen;WANG Yuchen;LIU Jun;YUAN Li;PAN Zhifen;ZHANG Lu(State Key Laboratory of Genetic Engineering,School of Life Sciences,Fudan University,Shanghai 200438,China;Shihezi University School of Medicine,Shihezi 832002,Xinjiang,China;Department of Tuberculosis,the First Hospital of Jiaxing,Jiaxing 314000,Zhejiang,China)

机构地区：[1]复旦大学生命科学学院遗传工程国家重点实验室,上海200438 [2]新疆石河子大学医学院,新疆石河子832002 [3]嘉兴学院附属第一医院嘉兴市第一医院结核科,浙江嘉兴314000

出　　处：《检验医学》2020年第6期614-619,共6页Laboratory Medicine

基　　金：“十三五”国家科技重大专项(2017ZX10201301-005);嘉兴市科技计划项目(2018AD32089)。

摘　　要：目的探讨结核分枝杆菌(MTB)Fad D9重组蛋白在活动性肺结核诊断中的价值。方法制备Fad D9重组蛋白并进行鉴定,使用C57BL/6小鼠模型评估其免疫原性。以Ag85A蛋白为阳性参照抗原,检测145例活动性肺结核患者(活动性肺结核组)、38例结核患者密切接触者(MTB密切接触组)和101名体检健康者(正常对照组)血清样本中Fad D9重组蛋白的特异性抗体水平,将Fad D9重组蛋白作为抗原与58例肺结核患者、34例非肺结核患者的全血样本37℃共孵育20~24 h后,采用γ-干扰素释放试验(IGRA)检测,将检测结果与IGRA试剂盒中的标准管进行比对,并采用受试者工作特征(ROC)曲线分析Fad D9重组蛋白诊断结核病的敏感性和特异性。结果Fad D9重组蛋白可在C57BL/6小鼠体内引起较高水平的体液免疫反应,且与阳性参照抗原Ag85A接近。活动性肺结核组抗Fad D9抗体水平高于MTB密切接触组和正常对照组(P<0.0001),MTB密切接触组与正常对照组之间差异无统计学意义(P>0.05)。抗Ag85A抗体水平3组之间差异均无统计学意义(P>0.05)。ROC曲线分析结果显示,Fad D9重组蛋白诊断结核病的曲线下面积(AUC)为0.682,最佳临界值为0.343,敏感性为96.8%,特异性为37.5%。结论Fad D9蛋白在活动性结核病的诊断中具有一定的临床应用潜力。Objective To investigate the role of FadD9 recombinant protein of Mycobacterium tuberculosis(MTB)for the diagnosis of active tuberculosis.Methods FadD9 recombinant protein was prepared and identified.C57BL/6 mouse model was used to evaluate its immunogenicity.Using Ag85A protein as positive reference antigen,the specific antibody levels of FadD9 recombinant protein in serum samples of 145 active tuberculosis patients(active tuberculosis group),38 close-contact tuberculosis patients(MTB close-contact group)and 101 healthy subjects(healthy control group)were determined.The FadD9 recombinant protein was incubated with blood samples of 58 tuberculosis patients and 34 non-tuberculosis patients at 37℃for 20-24 h,and then was determined by interferongamma release assay(IGRA).The results were compared with the standard tube in IGRA kit,and the sensitivity and specificity of FadD9 recombinant protein in the diagnosis of tuberculosis were analyzed by receiver operating characteristic(ROC)curve.Results FadD9 recombinant protein could induce a high level of humoral immune response in mice,which was close to that of Ag85A.Serum anti-FadD9 antibody level in active tuberculosis group was higher than those in healthy control group and MTB close-contact group(P<0.0001).There was no statistical significance between MTB close-contact group and healthy control group(P>0.05).There was no statistical significance among the 3 groups in the level of anti-Ag85A antibody(P>0.05).For the diagnosis of tuberculosis,the area under curve(AUC)of FadD9 recombinant protein was 0.682,the optimal cut-off value was 0.343,and the sensitivity and specificity were 96.8%and 37.5%,respectively.Conclusions FadD9 recombinant protein is potential in the diagnosis of active tuberculosis.

关 键 词：FadD9重组蛋白 γ-干扰素释放实验 结核分枝杆菌 

分 类 号：R446.1[医药卫生—诊断学]