蓝舌病毒抗体量子点检测试纸条的研制

Development of a Quantum Dots-based Immunochromatographic Strip for Detection of the Antibodies against Bluetongue Virus

为建立一种快速检测蓝舌病病毒抗体的免疫层析方法,本研究通过量子点标记葡萄球菌A蛋白(SPA),以蓝舌病病毒VP7蛋白作为检测线,抗葡萄球菌A蛋白单克隆抗体作为质控线组装量子点免疫层析试纸条。结果显示,该量子点试纸条在蓝舌病病毒抗体阳性血清稀释度为1∶256时仍可检出;试纸条阳性阈值为0.0668,并且根据阈值确定了试纸条检测蓝舌病抗体最低含量为0.25 ng/mL;该试纸条与结核病、布鲁氏菌病、病毒性腹泻、巴氏杆菌病、小反刍兽疫及地方流行性白血病的阳性血清均无交叉反应;与国外商品化试剂盒相比较,该试纸条检测结果与ELISA检测试剂盒(IDEXX)的符合率为96.9%(63/65)。结果表明,量子点免疫层析试纸条在检测蓝舌病方面具有快速、灵敏度高、特异性强的优势,适用于蓝舌病的流行病学调查和临床快速筛查。

To develop a quantum dot immunochromatography test strip for rapidly detecting the antibodies against Bluetongue virus,the staphylococcus A protein was used for quantum dot marker and the recombinant protein VP7 was served as test line and the monoclonal antibody to staphylococcus A protein was served as control line.As a result,even standard bluetongue disease-positive sera diluted at 1:256 was detected.The positive threshold value of the test strip was 0.0668,and the minimum content of the antibody for detecting bluetongue disease by the test strip is 0.25 ng/mL.The prepared quantum dot strip showed no cross reaction with the sera from other diseases.Comparative study showed the coincidence of 96.9%between the test strip and commercial ELISA kit(IDEXX)(63/65).In conclusion,the quantum dot immunochromatographic test strip developed here might be used to quickly and accurately detect antibodies against Bluetongue virus and suitable for clinical diagnosis and epidemiological study.