时间分辨荧光免疫法定量测定孕妇血清中甲胎蛋白含量

Quantitative test of human alphafetoprotein in maternal serum by time-resolved fluoroimmunoassay

目的探讨孕妇血清甲胎蛋白(MSAFP)含量与孕周、孕妇体质量和孕妇年龄的关系。方法自建MSAFP时间分辨荧光免疫测定法(TRFIA)并对其进行评价,采用自建方法定量检测孕妇血清3711例,建立实验室参考值,分析AFP浓度与孕周、孕妇体质量和孕妇年龄的关系。结果自建AFP-TRFIA方法灵敏度优于1.0U/ml,线性范围为1.00～1000.00U/ml,分析内、分析间变异系数(CV)分别不超过10.0%、15.0%,与国外同类试剂比对结果相关性好(r=0.9856)。在13～13周+6d,14～14周+6d,15～15周+6d,16～16周+6d,17～17周+6d,18～18周+6d,19～19周+6d,20～20周+6d,21～21周+6d各孕周的MSAFP含量中位数(M)分别为16.51U/ml,23.81U/ml,29.98U/ml,32.96U/ml,36.86U/ml,43.01U/ml,49.61U/ml,59.66U/ml,71.11U/ml。各孕周MSAFP平均浓度或浓度中位值与孕周呈线性正相关,r分别为0.9766,0.9832。各体质量段MSAFP浓度与孕妇体质量呈负相关,r在-0.9929～-0.9142。同孕周组内,各年龄段间的MSAFP浓度均数差异无统计学意义(P>0.05)。结论自建AFP-TRFIA法检测性能能满足临床应用的需要。MSAFP浓度与孕周和孕妇体质量密切相关,在产前筛查时要准确校对以上因素。

Objective To develop a time-resolved fluoroimmunoassay（TRFIA） for maternal serum alphafetoprotein （MSAFP） as an aid in prenatal screening,evaluate the impact of gestational weeks,maternal weight,gestational ages on MSAFP levels.Methods The assay methodology of self-made method was evaluated.The level of AFP was quantitative tested by self-made method in 3711 healthy maternal serum samples.The reference value of normal was studied.Statistics was used to analyze the impact of gestational weeks,maternal weight,gestational ages on MSAFP levels.Results The sensitivity of self-made method was better than 1.00 U/ml.The linear range was 1.00 U/ml to 1000.00 U/ml.The intra-assay and inter-assay coefficients of variation（CV） was lower than 10% and 15%,respectively.The accordance of our kit with the imported kit was good（P0.05）,and the accordance coefficient was 0.9856.The median of MSAFP level was 16.51,23.81,29.98,32.96,36.86,43.01,49.61,59.66,71.11 U/ml in 13-13＋6 d,14-14＋6 d,15-15＋6 d,16-16＋6 d,17-17＋6 d,18-18＋6 d,19-19＋6 d,20-20＋6 d,21-21＋6 d gestational week,respectively.From the 13th week to the 21th week,the MSAFP levels showed positive line correlation with gestational weeks.The MSAFP levels showed negative correlation with maternal weight,but the MSAFP levels showed no correlation with maternal ages,there was no statistical significance for MSAFP average levels with gestational ages in the group of ages （P0.05）.Conclusions All the results of the parameters showed that the method we developed could meet the need of the clinicapplication.Gestational weeks,maternal weight bears directly on the levels of MSAFP for prenatal screening.