应用酶联免疫斑点法快速诊断儿童结核分枝杆菌感染的研究

目的:探讨基于ESAT-6和CFP-10抗原肽的酶联免疫斑点技术(Enzyme-link immunospot essay,ELISPOT)在临床实践中诊断儿童结核分枝杆菌感染的临床价值。方法:应用ELISPOT技术检测14例确诊的儿童肺结核病例和16例健康儿童对照的外周血单个核细胞的INF-γ,同时对所有研究对象实施结核菌素皮肤实验(Tuberculin Skin Test,TST),探讨ELIS-POT技术在快速诊断儿童结核分枝杆菌感染的临床应用价值。结果:ELISPOT灵敏度为85.7%(95%CI:60.1～95.0),特异度为93.8%(95%CI:71.7～98.9)。TST灵敏度为71.4%(95%CI:45.4～88.3),特异度为56.3%(95%CI:33.2～76.9)。ELISPOT的灵敏度高于TST,但无统计学意义(P>0.05)。ELISPOT的特异度显著高于TST(P<0.05)。ELISPOT和TST检测结果总一致性较差(κ=0.35)。在活动型结核病组中,ELISPOT和TST一致性较差(κ=0.18),类似地,在对照组中,ELIS-POT和TST一致性也较差(κ=0.16)。不一致的检测结果在病例组中常见ELISPOT阳性而TST阴性(3例),而在对照组中常见于ELISPOT阴性而TST阳性(7例)。结论:ELISPOT诊断儿童活动性结核病准确性高于TST。ELISPOT具有较高的诊断灵敏度和特异度,其特异度不受儿童的卡介苗接种的影响,适合作为临床诊断儿童结核分枝杆菌感染的辅助诊断技术,有助于感染的儿童得到更快速的诊断和治疗。

Study on the Enzyme-link immunospot essay in diagnosis of Mycobacterium tuberculosis infection in children

Objective:To evaluate the usefulness of enzyme-link immunospot essay(ELISPOT) based on Earlysecreting antigen target-6(ESAT-6) and(culture filtrate protein-10,CFP-10) in the diagnosis of Mycobacterium tuberculosis infection in children.Methods:The 14 children with active tuberculosis and 16 healthy control were enrolled.The peripheral blood mononuclear cells of all participants were detected by ELISPOT,and all participants were also performed Tuberculin Skin Test(TST) simultaneously.Results:The sensitivity and specificity of ELISPOT for active TB were 85.7%(95%CI:60.1-95.0) and 93.8%(95%CI:71.7-98.9)respectively,while the above values of TST were 71.4%(95%CI:45.4-88.3)and 56.3%(95%CI:33.2-76.9)respectively.The sensitivity of ELISPOT was higher than that of TST,although there was no statistically significant.The specificity of ELISPOT was significantly higher than that of TST(P<0.05).The total agreement between ELISPOT and TST was poor(κ=0.35).The agreement between ELISPOT and TST in active TB cases and healthy control were poor(κ=0.18 and 0.16 respectively).Discordance in cases was most often ELISPOT positive and TST negative,nevertheless,discordance in healthy control was most often ELISPOT negative and TST positive.Conclusion:The validity of ELISPOT is higher than TST.ELISPOT has high sensitivity and specificity which may not influenced by the Bacille Calmette-Guérin(BCG) vaccination.ELISPOT can take as the supplementary role for the diagnostic of Mycobacterium tuberculosis Infection.It is helpful for the rapid diagnosis and treatment for the infectious children.