新城疫病毒和禽流感病毒复合型胶体金免疫层析试纸条的制备

为了建立一种简便、快速而且能同时检测新城疫病毒(NDV)和禽流感病毒(AIV)的方法,本试验采用柠檬酸三钠还原法制备胶体金颗粒,分别标记NDV单克隆抗体6C4和AIV单克隆抗体制备免疫检测探针。在硝酸纤维素膜上,用微定量喷头喷好2条病毒检测线(T线)和1条羊抗鼠抗体质控线(C线),制备复合型免疫层析检测试纸条。结果在10 min内,可同时检测出两种病毒。试纸条检测NDV的灵敏度比HA试验结果提高8倍;AIV重组抗原的检测灵敏度为1.7μg/mL。两种病毒互相测试,未出现交叉反应。用缓冲液对照测试结果为阴性。在密封、干燥、低温的条件下,试纸条的灵敏性与特异性没有明显变化。说明本研究建立的NDV和AIV免疫层析检测法具有特异、灵敏、稳定、操作简单等特点,符合现场快速检测的要求。

Development of a complex gold-immunochromatography test strip for newcastle disease virus and avian influenza virus

To develop a simple,rapid and highly efficient immunochromatographic assay for newcastle disease virus(NDV) and avian influenza virus(AIV),the sodium citrate reduction was used to make colloidal gold particles which will conjugate with NDV and AIV monoclonal antibodies,respectively,as immunoassay probes.The test lines(T) for the two viruses were sprayed on the nitrocellulose membrane,while a sheep anti-mouse antibody quality control line(C) was sprayed.The strip could detect two viruses simultaneously in ten minutes.The detection strip of NDV was more sensitive than the HA test by improving 8 times,and the recombinant antigen of AIV could still be detected when diluted to 1.7 μg/mL.Two viruses had no cross-reactions with each other.With the buffer as control,the result was negative.The sensitivity and specificity of strips had not changed when stored under a sealed,dry,low temperature condition.The results revealed that the strip for detection of NDV and AIV has high specificity,sensitivity and stability,and is suitable for rapid detection of the two viruses in the field.