时间分辨荧光免疫法风疹病毒IgG抗体定量测定试剂盒的研制与性能评价

Development and performance evaluation of time-resolved fluorescence immunoassay quantitative kit on determination of rubella virus IgG antibody

目的建立风疹病毒（RV）IgG抗体时间分辨荧光免疫法（TrFIA）并研制其试剂盒。方法采用RV抗原作为包被抗原，铕标记羊抗人IgG作为示踪物，配制以β一萘甲酰三氟丙酮为主要成分的增强液，应用TrFIA法测定RVIgG，并对自研试剂盒的最低检测限、准确度、线性、重复性、阴性参考品符合率、阳性参考品符合率、热稳定性进行评价，与同类试剂盒进行方法学比对试验，比对试验定量结果的等效性采用线性回归分析。结果自研试剂盒的最低检测限、重复性、阴性参考品符合率、阳性参考品符合率均能达到国家检定标准；检测国际参考品（10IU／mL）或企业校准品，其实测值与理论值相对偏差均在正负20．0％以内；在2．0～256．0IU／mL范围内，线性相关系数不低于0．9900；于37℃恒温箱放置6d后，检测性能无明显改变；临床研究评价一致程度百分比可达100％。结论自研试剂盒灵敏度高、特异性强、精密度好、准确度高、线性范围宽，与同类产品检测结果相关性好，能满足临床需要。

Objective To establish the rubella virus （RV） IgG antibody time-resolved fluorescence immunoassay（TrFIA） and the development of test kit. Methods RV antigen was taken as coating antigen,europium-labeled goat anti-human IgG was taken as a tracer, we constructed a TrFIA kit and the self-developed kit was evaluated by testing the lowest limit of detection,accuracy, line- arity,and repeatability. Results The kit＇s detection limit,repeatability are in line with the rate of negative reference materials. The measured value is highly proportioned to the theoretical value;within 2.0-256.0 IU / mL range, the linear correlation coefficient of higher than 0. 990 0. Conclusion The self-veloped detection kits provide satisfactory sensitivity,specificity,precision,and accura- cy,and it can be of help in clinical detection.