【摘要】 目的 评价酶联免疫斑点(ELISPOT)试验在脊柱结核诊断中的应用价值。方法 选择2013年5月至2017年1月在安徽省胸科医院确诊为脊柱结核的患者98例(脊柱结核组)和非结核病患者77例(非结核组)。采用ELISPOT检测脊柱结核组和非结核组外周血单个核细胞,观察斑点形成细胞的个数;患者同时接受结核菌素皮肤试验(PPD)及血结核抗体(TB-Ab)检测,其中PPD皮肤试验分别采用红肿硬结平均直径≥5mm和≥15mm两种标准,标记为PPD-5和PPD-15,并对比分析以上检测方法的敏感度、特异度、阳性预测值和阴性预测值。结果ELISPOT在脊柱结核诊断中的敏感度、特异度、阳性预测值、阴性预测值分别为93.88%(92/98)、92.21%(71/77)、93.88%(92/98)和92.21%(71/77)。PPD-5和PPD-15皮肤试验的敏感度、特异度、阳性预测值、阴性预测值分别为86.73%(85/95)和52.04%(51/98)、74.03%(57/77)和89.61%(69/77)、80.95%(85/105)和86.44%(51/59)、81.43%(57/70)和59.48%(69/116)。血TB-Ab检测的敏感度、特异度、阳性预测值和阴性预测值分别为48.98%(48/98)、61.04%(47/77)、61.54%(48/78)和48.45%(47/97)。血ELISPOT的敏感度、阴性预测值高于PPD-15试验(χ~2=43.47,P<0.01;χ~2=28.65,P<0.01);特异度、阳性预测值、阴性预测值高于PPD-5试验(χ~2=9.07,P<0.01;χ~2=7.45,P<0.01;χ~2=4.98,P=0.026);敏感度、特异度、阳性预测值、阴性预测值均高于血TB-Ab试验(χ~2=48.40,P<0.01;χ~2=20.88,P<0.01;χ~2=29.59,P<0.01;χ~2=44.97,P<0.01),以上差异均有统计学意义。非结核组标本中早期分泌抗原靶蛋白6(ESAT-6)和培养滤液蛋白10(CFP-10)特异性T淋巴细胞分泌γ干扰素的淋巴细胞斑点数[中位数(四分位数),M(Q1,Q3)]分别为0(0,3)和0(0,2)个,脊柱结核组标本中ESAT-6和CFP-10特异性T淋巴细胞分泌γ干扰素的淋巴细胞斑点数分别为29.00(9.57,57.75)和20.00(7.50,73.75)个,脊柱结核组均高于非结核组,差异均有统计学意义(U值分别为492.00和427.00,P值均<0.01)。结论 ELISPOT应用于脊柱结核的诊断有良好的敏感度和特异度,可作为脊柱结核感染辅助诊断的有效方法。更多还原

【Abstract】 Objective To evaluate the performance of enzyme-linked immunospot(ELISPOT)assay in the diagnosis of spinal tuberculosis(TB). Methods Participants included 98 cases diagnosed with spinal TB and77 patients with non-TB in Anhui Chest hospital from May.2013 to Jan.2017.The peripheral blood mononuclear cells were collected from all subjects in the spinal TB group and non-TB group and analyzed by ELISPOT.The number of spot forming cells(SFCs)was counted.All patients also received purified-protein derivative(PPD)tuberculin skin test and serum tuberculosis antibody(TB-Ab)detection.For the PPD skin test,swelling and induration diameter of≥5 mm and≥15 mm were used as two standards for positive criteria,respectively,marked as PPD-5 and PPD-15.The sensitivity,specificity,positive predictive value and negative predictive value of all tests were analyzed and compared. Results The sensitivity,specificity,positive predictive value and negative predictive value were 93.88%(92/98),92.21%(71/77),93.88%(92/98)and 92.21%(71/77)for ELISPOT analysis,86.73%(85/95),74.03%(57/77),80.95%(85/105)and 81.43%(57/70)for PPD-5 skin test,52.04%(51/98),89.61%(69/77),86.44%(51/59),and 59.48%(69/116)for PPD-15 skin test,48.98%(48/98),61.04%(47/77),61.54%(48/78)and 48.45%(47/97)for TB-Ab test,respectively.The sensitivity and negative predictive value of ELISPOT blood test were significantly higher than those of the PPD-15 test(χ~2=43.47,P<0.01;χ~2=28.65,P<0.01).The specificity,positive predictive value and negative predictive value were significantly higher than those of the PPD-5 test(χ~2=9.07,P<0.01;χ~2=7.45,P<0.01;χ~2=4.98,P=0.026).The sensitivity,specificity,positive predictive value and negative predictive value of ELISPOT assay were significantly higher than those of the TB-Ab blood test(χ~2=48.40,P<0.01;χ~2=20.88,P<0.01;χ~2=29.59,P<0.01;χ~2=44.97,P<0.01).The numbers of SFCs for early-secreted antigenic target protein 6(ESAT-6)and 10 kD culture filtrate antigen(CFP-10)were 0(0,3)and 0(0,2)(M(Q1,Q3))in the spinal TB group,and 29.00(9.57,57.75)and20.00(7.50,73.75)in the non-TB group;differences between the two groups were statistically significant(U=492.00 and 427.00,respectively;P<0.01). Conclusion ELISPOT assay in spinal TB diagnosis has good sensitivity and specificity,and can be used as an effective method for auxiliary diagnosis of spinal TB infection.更多还原