D-二聚体胶体金免疫层析定量检测方法的初步建立

Establishment of the quantitative detection method of colloidal gold immunochromatography for D-Dimer

目的建立一种检测D-二聚体的胶体金免疫层析定量检测方法,为临床检测D-二聚体提供一种简便、快速、准确的检测方法。方法根据免疫学中双抗体夹心和免疫层析的原理,研制定量检测样本中D-二聚体的胶体金试纸条,在金标定量仪上建立标准曲线,从灵敏度、特异性、线性、精密性、稳定性和方法学对比等方面对该方法进行评价。结果本试纸条的线性范围为0.156 25~10μg/ml,回归方程Y=92.907 X+307.88,R2=0.982 68;批内变异系数(CV)为3.78%和2.81%,批间变异系数(CV)为4.95%和4.07%;与纤维蛋白原无交叉反应性;在37℃条件下保存1周;与IL公司ACL-TOP700检测系统免疫比浊法的相关性良好,回归方程Y=0.959 64 X+0.026 45,相关系数R2=0.985 93。结论本研究初步建立了检测D-二聚体的胶体金免疫层析定量检测方法,为临床检测D-二聚体提供了一种新的技术和方法。

Objective To establish a quantitative detection method based on the principle of immune chromatography,which provides a new detection method for detecting D-Dimer that is fast,accurate,simple and convenient. Methods According to the principle of double antibody sandwich and immune chromatography in immunology,a quantitative detection of the colloidal gold test strip of D-Dimer was created. And standard curve on gold standard quantitative instrument have been established. Besides,the method was evaluated by the sensitivity,specificity,linearity,precision,stability and methodology comparison.Results The linear range was 0. 156 25 ~ 10 μg / ml,and the regression equation was Y = 92. 907 X + 307. 88,R2= 0. 982 68.The intra-group variation coefficients were 3. 78% and 2. 81%,and the inter-group variation coefficients were 4. 95% and4. 07%. Otherwise,the test strip had no cross reactivity with fibrinogen and the stability. The test strips could be stored for144 hat 37 ℃. The correlation between the method we have established and the immunoturbidimetry of IL company was good,and the regression equation was Y = 0. 959 64 X + 0. 026 45,R2= 0. 985 93. Conclusion In this study,we have initially and successfully established the colloidal gold immune chromatography assay for the quantitative detection of D-Dimer. We provide a new technique and method for clinical detection of D-Dimer.