血清甲胎蛋白和游离β绒毛膜促性腺激素双标记时间分辨荧光免疫试剂盒的研制与评价

摘要

Objective The aim of the study was to promote the development of the prenatal screening,and to develop a dual-label time-resolved fluoroimmunoassay (TrFIA) kit for serum alpha-fetoprotein (AFP)and free human chorionic gonadotropin β subunit (free hCG β) as an aid in prenatal screening. Methods The kit was based on solidphase sandwich TrFIA for simultaneous detection of alpha-fetoprotein and the free human chorianic gonadotropin β suburtit,which involved two trances,One was anti-AFP monoclonal antibody europium marker,the other was anti-Free hCG β monoclonal antibody samarium marker. The main component of enhancement solution was 2-naphthalene thrifluoroacetone. The assay methodology was evaluated. Linear regression analysis was used. Results The correlation coefficient of AFP and free hCG β doseresponse curve could be reached 0. 999 0. The intra-assay and inter-assay coefficients of variation (CV) was lower than 10％. The sensitivity of AFP and free hCG β were 0. 12 U/ml and 0. 08 ng/ml,respectively. The relative potency of national calibrators to the kit calibrators were 0. 900～1. 100 for testing AFP and free hCG β. The linear range was 0. 25 U/ml ～500 U/ml for testing AFP. The linear range was 0.2 ng/ml～200 ng/ml for testing free hCG β. The stable period of reagents was 12 months by storing at 4 ℃. Compared with import kit,the correlation coefficient of AFP and free hCG β were 0. 973 0 and 0. 992 7,respectively. Conclusion The self-developed kit has high sensitivity,specificity,accuracy,wide linear range,and it's performance indicators is as well as import kit. The self-developed kit is valuable for clinical application.%目的 研制一种用于产前筛查的母血清甲胎蛋白(AFP)和游离β绒毛膜促性腺激素(free hCG β)双标记时间分辨荧光免疫法(TrFIA)试剂盒.方法 采用AFP单克隆抗体铕(Eu3+)标记物和free hCGβ单克隆抗体钐(Sm3+)标记物作为示踪物,以β-萘甲酰三氟丙酮为主要成分制备增强液,应用双抗体夹心法研制出hAFP和free hCG β双标记时间分辨荧光免疫法试剂盒,并对其进行分析性能评价,平行比对试验采用线性回归分析.结果 自制试剂盒AFP和free hCG β的剂量-反应曲线线性相关系数(r)可达0.999 0以上;批内、批间CV(%)均小于10.00%;AFP和free hCG β的灵敏度分别为0.12 U/ml和0.08 ng/ml;以国家标准品为对照,AFP和free hCG β的标准品效价比在0.900～1.100间;检测AFP和free hCG β的线性范围分别为0.25 U/ml～500 U/ml和0.2 ng/ml～200 ng/ml;试剂盒在4℃保存下可至少稳定12月;与国外同类试剂比对,AFP和free hCG β的回归方程的线性相关系数(r)分别为0.973 0和0.992 7.结论 自制试剂盒具有灵敏度高、特异性强、准确度好、线性范围宽等优点,与国外的同类产品检测结果相关性好,能满足临床需要.