T-SPOT联合颗粒酶B阳性Ts淋巴细胞对成人结核病诊断的价值  

The Value of T-SPOT Combined with Granzyme B-positive Ts Lymphocytes for the Diagnosis of Tuberculosis in Adults

作　　者：戴乾滨 陈桉军 朱清[1] 魏丹丹[1] 刘蓬 刘洋[1] DAI Qianbin;CHEN Anjun;ZHU Qing;WEI Dandan;LIU Peng;LIU Yang(Department of Laboratory,the First Affiliated Hospital of Nanchang University,Nanchang 330006,China)

机构地区：[1]南昌大学第一附属医院检验科,江西南昌330006

出　　处：《标记免疫分析与临床》2024年第8期1454-1458,1534,共6页Labeled Immunoassays and Clinical Medicine

基　　金：国家自然科学基金地区项目(编号:82260403);江西省卫生计生委科技计划项目(编号:202310458)。

摘　　要：目的探讨结核感染T细胞斑点试验(T-SPOT.TB)联合颗粒酶B(GrB)+CD8^(+)T淋巴细胞在诊断结核病中的价值。方法选取南昌大学第一附属医院2021年2月至2022年3月疑似结核病患者212例,所有患者均保留T-SPOT结果,流式细胞仪检测外周血中T淋巴细胞亚群及其穿孔素和GrB的表达,根据出院最终诊断分为结核病组和非结核病组,比较两组间T-SPOT斑点数和淋巴细胞亚群及其穿孔素和GrB表达差异,ROC曲线分析ESAT-6斑点数,CFP-10斑点数,结核特异性抗原(TBAg)与植物血凝素(PHA)斑点数的比值(TBAg/PHA)(ESAT-6/PHA和CFP-10/PHA中较大的一个),GrB^(+)Ts(CD8^(+)T)及“结核比值”[TBAg/(PHA*GrB^(+)Ts)]的诊断效能。结果GrB^(+)Ts细胞百分比在结核组中显著高于非结核组(44.95±2.98 vs 33.49±1.87,P<0.05),而总T细胞、穿孔素+T细胞,GrB^(+)T细胞,Ts细胞和穿孔素+Ts细胞百分比差异无统计学意义(P>0.05);非结核组T-SPOT.TB阳性率为51.85%,明显低于结核组的92%,且结核组的A抗原斑点数及B抗原斑点数显著大于非结核组(P<0.05),ROC曲线显示A抗原诊断结核病的灵敏度和特异性分别为80.0%和56.8%,曲线下面积(AUC)为0.748,约登指数为0.368;B抗原的灵敏度和特异性为分别62.0%和77.8%,AUC为0.758,约登指数为0.398;A抗原+B抗原灵敏度和特异性为72.0%和74.1%,AUC为0.796,约登指数为0.461;结核组的TBAg/PHA比率要显著高于非结核组(1.21±0.17 vs 0.26±0.02,P<0.001)以0.59为cut-off值时,诊断结核病的灵敏度和特异性分别为66%和91.4%,AUC高达0.852,约登指数为0.574;结核组的“结核比值”要显著高于非结核组(4.58±0.87 vs 1.71±0.24,P<0.01),以1.24为cut-off值时,诊断结核病的灵敏度和特异性分别为82.0%和72.2%,AUC为0.794,约登指数为0.542。结论应用TBAg/PHA比率和TBAg/(PHA*GrB^(+)Ts)比值诊断结核病的效能要优于常规的T-SPOT检测,两者结合使用对结核病诊断可能具有重要价值。Objective To investigate the value of T-cell spot test for tuberculosis infection(T-SPOT.TB)combined with granzyme B^(+)(GrB^(+))CD8^(+)T lymphocytes in the diagnosis of tuberculous disease.Methods 212 patients with suspected tuberculous disease from February,2021 to March,2022 in the First Affiliated Hospital of Nanchang University were selected for the study,all of whom were retained for T-SPOT results.T lymphocyte subpopulations and expressions of perforin and granzyme B were detected in peripheral blood by flow cytometry.These patients were then divided into the tuberculosis and non-tuberculosis groups based on final diagnosis at hospital discharge.The number of T-SPOT spots and lymphocyte subpopulations and their perforin and granzyme B expressions were compared between the two groups.ROC curves were drawn to evaluate the diagnostic efficacy of ESAT-6 spot number,CFP-10 spot number,TBAg/PHA ratio(the greater of ESAT-6/PHA and CFP-10/PHA),granzyme B^(+)Ts(CD8^(+)T),and the“TB Ratio”[TBAg/(PHA*Granzyme B^(+)Ts)].Results The percentage of GrB^(+)Ts cells was significantly higher in the TB group than in the non-TB group(44.95±2.98 vs 33.49±1.87,P<0.05),whereas there was no difference in the percentage of total T cells,perforin+T cells,GrB^(+)T cells,Ts cells and perforin+Ts cells(P>0.05);The T-SPOT.TB positivity rate in the non-TB group was 51.85%,which was significantly lower than the value of 92%in the TB group.The numbers of A-antigen spots and B-antigen spots in the TB group were significantly greater than those in the non-TB group(P<0.05).ROC curve showed that the sensitivity and specificity of A-antigen for diagnosing TB were 80%and 56.8%,respectively,with an area under the curve(AUC)of 0.748,and that of B-antigen were 62.0%and 77.8%,respectively,while AUC was 0.758;The sensitivity and specificity of A+B antigen were 72.0%and 74.1%with an AUC of 0.796;The TBAg/PHA ratio in the TB group was significantly higher than that in the non-TB group(1.21±0.17 vs 0.26±0.02,P<0.001).The sensitivity and specifi

关 键 词：结核 T-SPOT TBAg/PHA比率 TBAg/(PHA*Ts颗粒酶B)比值 

分 类 号：R52[医药卫生—内科学]