摘要
目的研铺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（freehCGB）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（Eu^3＋）标记物和freehCGis单克隆抗体钐（Sm^3＋）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freehCG卢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。结果自制试刺盒AFP和freehCGβ的剂量-反应曲线线性相关系数（r）可达0．9990以上；批内、批间CV（％）均小于10．00％；AFP和freehCGP的灵敏度分别为0．12U／ml和0．08ng／ml；以国家标准品为对照，AFP和freehCG8的标准品效价比在0．900～1．100间；检测AFP和freehCG8的线性范围分别为0.25U／ml-500U／ml和0．2ng／ml-200ng／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freehCGβ的回归方程的线性相关系数（r）分剐为0．9730和0．9927。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。
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 chorionic gonadotropin β subunit,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 thrifluoroaeetone. 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.
出处
《现代检验医学杂志》
CAS
2011年第3期119-122,共4页
Journal of Modern Laboratory Medicine
关键词
甲胎蛋白
Β绒毛膜促性腺激素
铕
钐
时间分辨荧光免疫
alpha-fetoprotein
human chorionic gonadotropin β subunit
europium
samarium
time-resolved fluoroimmunoassay
