时间分辨荧光免疫法人巨细胞病毒IgG抗体定量测定试剂盒的研制

目的建立人巨细胞病毒IgG（HCMV IgG）抗体时间分辨荧光免疫法（TrFIA）并研制其试剂盒。方法采用HCMV pp150＋pp52＋pp65抗原作为包被抗原,铕标记羊抗人IgG作为示踪物,配制以β-萘甲酰三氟丙酮为主的增强液,应用间接TrFIA法测定HCMV IgG抗体,将自制试剂盒与同类试剂盒进行方法学比对试验,比对定量结果的等效性采用线性回归分析。结果自制HCMV IgG抗体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;检测标化的企业校准品,其实测值与理论值相对偏差均在±20.0%以内;在2.0 RU/mL~256.0 RU/mL范围内,线性相关系数不低于0.9900;于37℃恒温箱放置6天后,检测性能无明显改变;临床研究评价一致程度百分比达99.4%,1020例临床样本定量结果的线性回归方程为Y=0.9594X＋0.3252,r=0.9918。结论自制试剂盒测定HCMV IgG抗体灵敏度高、特异性强、精密度好、准确度高、线性范围宽;与同类检测结果相关性好,能满足临床需要。

Development of Time-resolved Fluoroimmunoassay for IgG Antibody of Human Cytomegalovirus

Objective To establish time-resolved fluoroimmunoassay（TrFIA） for IgG antibody of human cytomeg- alovirus（HCMV IgG）. Methods The HCMV pp150 ＋ pp52 ＋ pp65 antiage was coated on microplates, the goat anti-human IgG was labeled by Europrum. The enhancement solution was β-naphthalene thrifluoroacetone. The 1020 samples were analyzed by self-made kits and commercial kits. Results The detection limit, repeatability, compliance rate of negative and positive national standard of self-made TrFIA kit was in line with national accreditation standards. The linear range for HCMV IgG antibody of TrFIA was from 2.0 to 256.0 RU/mL, the linear correlation coefficient was up to 0. 9900. The analysis performance of kits had no significant changes after storing at 37℃ for six days. The testing results of self-made kit with reference kit for 1020 samples were linear correlation （ Y = 0. 9594X ＋ 0. 3252, r = 0. 9918）. Conclusion TrFIA for HCMV IgG antibody established in this study and self-made kit may be high sensitivity, specificity, good precision and high accuracy. It could meet clinical demand for the analysis of HCMV IgG antibody.