CA242荧光免疫层析定量检测试剂的建立

The Establishment and Application of Fluorescence Immunoassay Test Strip for CA242

目的为临床研制一种操作简易,灵敏度高的荧光免疫层析试纸条,用于定量检测CA242。方法采用双抗体夹心法和荧光免疫层析技术,将标记抗体L1与荧光微球结合于聚酯膜结合垫上,检测抗体C包被在硝酸纤维素膜上作为检测线(T),另将质控抗体Q1(羊抗鼠IgG)包被在相互平行的硝酸纤维素膜上作为质控线(C),过量的鼠源标记的荧光微球在质控线处聚集产生质控荧光信号,待测样中的CA242在T处形成荧光微球抗体-抗原-抗体的夹心复合物M2并产生荧光信号,通过荧光检测仪计算出定量结果。结果试纸条检测CA242灵敏度最低可达0.52kU/L,与CEA、CA125无交叉反应,批内CV为3.74%,批间CV为6.34%。与TrFIA方法比较相关性较好,R^2=0.9694;同一批试剂4℃放置6个月后,用100kU/L浓度的标准品进行测试,结果显示荧光计数CV为2.43%。结论该荧光免疫层析试纸条,可以提供较高的灵敏度和特异性,操作简单,能适合临床工作的需要。

Objective To establish a fluorescence immunoassay test strip(FITS)method for highly sensitive and convenient CA242 determination.Methods Labeled antibody L1 and fluorescent microsphere were conjugated to polyester membrane.Detecting antibody C1 and quality control antibody Q1(goat anti-mouse antibody)were coated on NC membrane separately,and named as the test(T)line and control(C)line,respectively.Mouse antibody labeled fluorescent microsphere was accumulated on C line,and at the same time,CA242 captured by L1 and C1 was accumulated on T line.The concentrations of CA242 were calculated by fluorescence detector.Results The sensitivity of FITS method was 0.52 kU/L.There were no cross reactivities with CEA and CA125.The within-run CV and between-run CV were 3.74%and 6.34%,respectively.The result of this method was well correlated with TrFIA(R^2=0.9694).Furthermore,the reagents from the same batch was stored at 4℃for 6 months before the standard samples with the concentration of 100 kU/L were tested,and the results showed that the fluorescence count CV was 2.43%.Conclusion The established FITS method from the current study has a relative high sensitivity and specificity,and can be user-friendly applied on CA242 clinical determination.