CAJ | 학술논문

目的：本研究建立基于量子点的志贺氏菌sFLISA检测方法并进行验证。方法以志贺氏菌多克隆抗体为包被抗体,以生物素标记的志贺氏菌单克隆抗体为第二抗体,以量子点标记的链酶亲和素进行荧光定量检测。结果试验确定志贺氏菌sFLISA检测最佳条件：包被抗体浓度为1.25μg/mL,生物素标记的单克隆抗体稀释倍数为1：400,量子点标记的链酶亲和素稀释倍数为1：100；特异性检验表明, sFLISA方法仅与志贺氏菌出现阳性反应,而与大肠埃希氏菌、沙门氏菌、葡萄球菌、李斯特氏菌、变形杆菌、副溶血弧菌、芽孢杆菌等均呈阴性反应。结论本研究建立的基于量子点的志贺氏菌sFLISA检测方法的最低检出量为3.5×104 CFU/mL,实现了对志贺氏菌高通量定性和定量检测。
목적：본연구건립기우양자점적지하씨균sFLISA검측방법병진행험증。방법이지하씨균다극륭항체위포피항체,이생물소표기적지하씨균단극륭항체위제이항체,이양자점표기적련매친화소진행형광정량검측。결과시험학정지하씨균sFLISA검측최가조건：포피항체농도위1.25μg/mL,생물소표기적단극륭항체희석배수위1：400,양자점표기적련매친화소희석배수위1：100；특이성검험표명, sFLISA방법부여지하씨균출현양성반응,이여대장애희씨균、사문씨균、포도구균、리사특씨균、변형간균、부용혈호균、아포간균등균정음성반응。결론본연구건립적기우양자점적지하씨균sFLISA검측방법적최저검출량위3.5×104 CFU/mL,실현료대지하씨균고통량정성화정량검측。
Objective To establish sFLISA (sandwich fluorescence-linked immunosorbent assay) method based on quantum dots and realize high-throughput detection ofShigellaspp.MethodsThe study used polyclonal antibody ofShigellaspp as the package antibody, and biotin-labeled clonal antibody ofShigella as the second antibody, and detected fluorescence by combination of QDs-labeled streptavidin and biotin.Results This study established sFLISA of ofShigella spp. based on quantum dots, and made validation testing using different bacteria strains. The research determined the optimum parameter ofShigella spp. in the end: the package antibody concentration was 1.25μg/mL, biotin-labeled clonal antibody diluted multiples was 1:400, and QDs-labled streptavidin diluted multiples was 1:100.ConclusionThe specificity testing showed that this method was positive only toShigella and negative to other bacteria strains. The sensitivity of the method was 3.5×104 CFU/mL.