布鲁氏菌量子点荧光微球免疫层析试纸条的研制

Development of a quantum dot fluorescent microspheres immunochromatographic test strip for Brucella detection

为制备基于量子点荧光微球的布鲁氏菌免疫层析试纸条,将布鲁氏菌脂多糖单克隆抗体和山羊抗鼠Ig G抗体分别作为检测线和质控线喷涂在硝酸纤维素膜上,以量子点荧光微球标记布鲁氏菌脂多糖单克隆抗体,稀释后喷涂在玻璃纤维素膜上,最后进行组装、切割制成检测用试纸条,并测定其敏感性、特异性和稳定性。以布鲁氏菌荧光定量PCR检测技术为对照,应用该试纸条检测临床样本120份,比较两种方法的符合率。结果显示,建立的量子点荧光微球免疫层析试纸条性能较好,检测敏感性达到1 ng/m L;特异性较好,与其他病原没有交叉反应;常温保存18个月或37℃保存28 d,其稳定性较好,变异系数分别为3.2%和14.6%;利用该试纸条和荧光定量PCR方法对120份临床样本进行检测,与荧光定量PCR检测结果相比,其阳性符合率为91%,阴性符合率为100%,总符合率为99%。上述结果表明,该试纸条操作简单、快速稳定、灵敏度高、成本低,20 min内即可完成检测,样本获取容易,可实现现场快速检测,在布鲁氏菌病的病原检测及净化工作中具有良好应用前景。

To prepare a Brucella immunochromatographic strip based on quantum dot fluorescent microspheres,Brucella lipopolysaccharide monoclonal antibodies and goat anti-mouse Ig G antibodies were sprayed onto nitrocellulose membranes as detection and quality control lines,respectively.Brucella lipopolysaccharide monoclonal antibodies were labeled with quantum dot fluorescent microspheres,diluted,and sprayed onto glass cellulose membranes.Finally,assembly and cutting were performed to produce test strips for detection,and their sensitivity,specificity,and stability were measured.Finally,using Brucella fluorescence quantitative PCR detection technology as a control,120clinical samples were tested using this test strip to compare the compliance rates of the two methods.Results,the established quantum dot fluorescent microsphere immunochromatographic test strip showed good performance,with a detection sensitivity of 1 ng/m L for brucellosis and good specificity.There was no cross reaction with other pathogens.The stability test found that the product had good stability when stored at room temperature for 18 months or 37℃for 28 d,with coefficients of variation of 3.2%and 14.6%,respectively;120 clinical samples were tested using test strips and fluorescence quantitative PCR methods.Compared with the results of fluorescence quantitative PCR,the positive conformity rate was 91%,the negative conformity rate was 100%,and the total conformity rate was 99%.Conclusion,the test strip has the advantages of simple operation,rapid stability,high sensitivity and low cost.The detection can be completed within 20 minutes,and the sample acquisition is easy.It can achieve rapid on-site detection and has good prospects in the pathogen detection and purification of brucellosis.