Wang Shwu-Jinng, Chang Wei-Feng, Wang Min-Ying, Hsiung Kuang-Pin, Liu Yung-Chuan
Department of Chemical Engineering, National Chung Hsing University, Taichung, Taiwan, ROC.
Vet Immunol Immunopathol. 2008 Oct 15;125(3-4):284-90. doi: 10.1016/j.vetimm.2008.05.022. Epub 2008 Jun 3.
For investigating the feasibility of using disperse dyes as an immunoassay chromogenic marker, a disperse dye, DADISPERSE NAVY BLUE SP, was selected in analyzing antibody against infectious bursal disease virus (anti-IBDV). With the color intensity revealed in the disperse dye immunochromatographic test (DICT) strip as the objective function, the optimal dyeing conditions were found as follows: dye concentration absorbance (at lambda(max)=587nm)=3, pH 7, 50 degrees C, for 10min. Under these conditions, the resultant dyed-antibody (rabbit anti-chicken) can produce an optimal color intensity reading of 55,054 on the strip. For performing qualitative immunoassay, chicken sera samples taken from different farms were used for the anti-IBDV titre assessment. The results of DICT strips showed very high sensitivity and specificity as compared to that analyzed by FlockChek enzyme linked immunosorbent assay (F-ELISA) kits. For quantitative immunoassay, it was found that the color intensity measured with DICT was linearly correlated to that of F-ELISA titre (r(2)=0.9687). Therefore, DICT was further applied to the detection of chicken anti-IBDV sera under vaccination in the farms. The average titres of the sampling groups exhibited a strong agreement to that of F-ELISA. Accordingly, the DICT method developed in this study, shown to be reliable, cheap and simple in both qualitative and quantitative immunoassays, is particularly suitable for point-of-need testing (PONT) in agricultural applications.
为研究使用分散染料作为免疫分析显色标记物的可行性，在分析传染性法氏囊病病毒抗体（抗IBDV）时选用了一种分散染料DADISPERSE NAVY BLUE SP。以分散染料免疫层析试验（DICT）条带中显示的颜色强度为目标函数，确定了最佳染色条件如下：染料浓度吸光度（在λ(max)=587nm处）=3，pH值7，50℃，持续10分钟。在这些条件下，所得的染色抗体（兔抗鸡）在条带上可产生55054的最佳颜色强度读数。为进行定性免疫分析，使用从不同养殖场采集的鸡血清样本评估抗IBDV效价。与FlockChek酶联免疫吸附测定（F-ELISA）试剂盒分析结果相比，DICT条带的结果显示出非常高的灵敏度和特异性。对于定量免疫分析，发现用DICT测量的颜色强度与F-ELISA效价呈线性相关（r(2)=0.9687）。因此，DICT进一步应用于养殖场中接种疫苗的鸡抗IBDV血清的检测。采样组的平均效价与F-ELISA的平均效价高度一致。据此，本研究开发的DICT方法在定性和定量免疫分析中均显示出可靠、廉价且简便的特点，特别适用于农业应用中的即时检测（PONT）。
