论文部分内容阅读
为建立一种新城疫病毒的双抗夹心液相芯片技术检测方法,将捕获抗体偶联到67号微球,对偶联抗体量、检测抗体和SA-PE抗体的工作浓度进行确定,用建立的液相芯片方法进行特异性、灵敏度、重复性及临床样品的检测。结果显示,1×10~6个微球的偶联抗体量为25 g,最优的检测抗体与SA-PE的工作浓度分别为2 g/mL和4 g/mL。该方法对其他病原无交叉反应,具有较好的特异性;检测的敏感效价为0.0625 HA,比传统的HA灵敏度高;批内、批间的变异系数分别为2.0%和6.7%;对60份临床样品检出率为5%,与PCR的检出率100%相符。结果表明,本试验建立的新城疫液相芯片检测方法具有特异性好、灵敏度高、重复性好等特点,为新城疫病毒的检测提供了一种新方法。
In order to establish a new method for the detection of Newcastle disease virus by double-antibody sandwich liquid-phase microarray, the capture antibody was coupled to No. 67 microsphere and the working concentration of conjugated antibody, detection antibody and SA-PE antibody was determined. Liquid Chips method for specificity, sensitivity, repeatability and detection of clinical samples. The results showed that the amount of conjugate antibody of 1 × 10 ~ 6 microspheres was 25 g, and the optimal concentrations of the detection antibody and SA-PE were 2 g / mL and 4 g / mL, respectively. The method has good specificity with no cross-reaction to other pathogens. The sensitivity of detection is 0.0625 HA, which is higher than the traditional HA. The coefficient of variation (CV) between intra-assay and inter-assay are 2.0% and 6.7% The detection rate of clinical samples was 5%, which was consistent with the detection rate of PCR 100%. The results showed that the Newcastle disease detection liquid phase chip established in this study was characterized by good specificity, high sensitivity and good repeatability, which provided a new method for the detection of Newcastle disease virus.