CAJ | 학술논문

目的：建立时间分辨荧光免疫分析法（TRFIA）检测结核分枝杆菌特异性6000早期分泌性抗原靶（ESAT-6）和抗原培养滤液蛋白10（CFP10）的融合抗原，并对该方法诊断肺结核的性能进行评价。方法采用夹心法建立ESAT-6-CFP10的TRFIA法。并对其灵敏度、批内、批间差异和准确性等各项指标进行考核。试验组选取24例活动性、32例可疑活动性、65例非活动性结核病患者及419例非结核性肺部疾病患者；30名健康体检者作为对照组，比较TRFIA法与酶联免疫吸附试验（ELISA）法检测结果。进行临床检测并与临床诊断进行一致性检验分析。结果 ESAT-6/CFP10的TRIFA法稳定性好，线性宽，其检测灵敏度为2.5 ng/mL，批内和批间变异系数分别平均为3.37%和5.23%，平均回收率为97.5%。与商业ELISA试剂盒一致性较好（Kappa=0.914，P=0.01）。 TRFIA法的剂量-反应曲线拟合相关系数（R2）达0.9961，可测范围为（2.5~1600） ng/mL；该方法与临床诊断有较好的一致性（Kappa=0.961，P=0.001）。结论 TRFIA法敏感度、精密度、重复性、线性相关性较好，有利于该结核抗原用于临床结核病的诊断。
목적：건립시간분변형광면역분석법（TRFIA）검측결핵분지간균특이성6000조기분비성항원파（ESAT-6）화항원배양려액단백10（CFP10）적융합항원，병대해방법진단폐결핵적성능진행평개。방법채용협심법건립ESAT-6-CFP10적TRFIA법。병대기령민도、비내、비간차이화준학성등각항지표진행고핵。시험조선취24례활동성、32례가의활동성、65례비활동성결핵병환자급419례비결핵성폐부질병환자；30명건강체검자작위대조조，비교TRFIA법여매련면역흡부시험（ELISA）법검측결과。진행림상검측병여림상진단진행일치성검험분석。결과 ESAT-6/CFP10적TRIFA법은정성호，선성관，기검측령민도위2.5 ng/mL，비내화비간변이계수분별평균위3.37%화5.23%，평균회수솔위97.5%。여상업ELISA시제합일치성교호（Kappa=0.914，P=0.01）。 TRFIA법적제량-반응곡선의합상관계수（R2）체0.9961，가측범위위（2.5~1600） ng/mL；해방법여림상진단유교호적일치성（Kappa=0.961，P=0.001）。결론 TRFIA법민감도、정밀도、중복성、선성상관성교호，유리우해결핵항원용우림상결핵병적진단。
Objective To establish a time-resolved fluorimmunoassay (TRFIA) for detecting the ESAT-6/CFP10 antigen and evaluate its clinic application. Methods A sandwich TRFIA of ESAT-6/CFP10 was established. The samples of ESAT-6/CFP10 in 24 patients with active tuberculosis, 32 patients with suspected active tuberculosis, 65 patients with inactivity tuberculosis, 419 non-tuberculous patients and 30 normal controls were measured. The sensitivity, precision and repetition of TRFIA were detected. The value of ESAT-6/CFP10 in the diagnosis mycobacterium tuberculosis infection were evaluated by using the consistency check of diagnostic test. Results The correlation coefficient (r2) of TRFIA dose-response curve reached 0.996 1. The detectability of detecting ESAT-6/CFP10 was 2.5 ng/mL, and the measurement range was (2.5-1 600) ng/mL. The inter and intra assay CVs were 3.37% and 5.23%, respectively. The recovery rate was 97.5%. TRFIA had high-level consistency with ELISA and clinical diagnosis(Kappa=0.914, P=0.001;Kappa=0.961, P=0.001; respectively). Conclusion The established sandwich TRFIA for ESAT-6/CFP10 measurement is a practicable assay. The results of precision, repetition, linear correlation and preliminary clinic application are satisfactory. It is helpful to diagnose mycobacterium tuberculosis infection with detecting ESAT-6/CFP10.