口蹄疫病毒抗原胶体金试纸条的研究

Study on Colloidal Gold Strip to Detect Foot-and-mouth Disease Virus

利用单克隆抗体技术制备抗口蹄疫病毒的单克隆抗体,特异性试验表明其只与O、A、Asia 1型3种血清型FMDV抗原结合。进而采用胶体金标记技术,以胶体金标记的抗口蹄疫病毒单克隆抗体、多克隆血清抗体和葡萄球菌A蛋白为主要材料,研制口蹄疫快速检测试纸条。该试纸条检测O、A、Asia 1型3种血清型灭活口蹄疫病毒均为阳性,检测水疱性口炎病毒、猪水疱病病毒、蓝舌病病毒、猪蓝耳病病毒4种灭活抗原及小反刍兽疫病毒疫苗株均为阴性,试验结果与口蹄疫实时荧光定量RT-PCR方法的完全一致,表明其具有良好的特异性。敏感性试验结果是,试纸条的检测极限为1∶160稀释的样品,其敏感性相当于实时荧光定量RT-PCR方法的1/64。由于试纸条具有操作方便、检测快速等优点,因此该试纸条可以用于大量临床样品的快速检测和现场检测。

In this study,a colloidal gold strip was developed for the detection of inactivated foot and mouth disease virus（FMDV） serotype A,O and Asia 1.Monoclonal antibodies against FMDV labeled with colloidal gold particles were used as a specific detector reagent to capture FMDV in samples.The purified anti FMDV polyclonal antibodies and staphylococcal protein A were immobilized on test and control regions of a nitrocellulose membrane,respectively.The colloidal gold strip was used to detect 8 inactivated viruses,including FMDV serotype A,O and Asia 1,Vesicular stomatitis virus,Swine vesicular disease virus,Bluetongue virus,Porcine reproductive and respiratory syndrome virus and Peste des petits ruminant virus.The results were same with real-time RT-PCR,which suggested the colloidal gold strip has good specificity.For serious diluted inactivated FMDV,the detecting limit of the colloidal gold strip was 160-fold diluted samples.The sensitivity was as 1/64 as that of real-time RT-PCR.As the colloidal gold strip has good specificity and sensitivity,and it is rapid and easy to use,the test is suitable for the rapid detection of FMDV.