应用基于上转换发光技术的免疫层析法检测鼠疫耶尔森氏菌

发布时间：2018-01-08 22:32

本文关键词：应用基于上转换发光技术的免疫层析法检测鼠疫耶尔森氏菌　出处：《中国人民解放军军事医学科学院》2006年硕士论文　论文类型：学位论文

【摘要】：目的：利用基于上转换发光技术(Up-Converting Phosphor Technology，UPT)生物传感器与免疫层析分析方法(immunochromatographic assay，ICA)相结合的技术平台，研制开发基于上转换发光材料(Up-Converting Phosphor，UCP)为标记物的免疫层析试纸条，实现对鼠疫耶尔森氏菌高灵敏度快速地现场检测，同时为UPT-ICA在多种生物分析中的广泛应用奠定基础。方法：采用400-nm大小的UCP颗粒作为标记物，双抗体夹心法作为反应模式来制备免疫层析试纸条；通过UPT生物传感器对免疫层析试纸条上经免疫反应而结合的UCP发光信号进行扫描检测，在一定范围内搜索检测带信号(V_T)和质控带信号(V_C)，并计算得出检测带与质控带的信号比值(V_T／V_C)，利用此信号比值可以对标本中的鼠疫耶尔森氏菌F1抗原、全菌进行定性和定量检测。结果：检测鼠疫耶尔森氏菌的UPT-ICA试纸条具有较高的特异性，检测灵敏度为10，，000CFU／ml，并在10~4～10~8CFU／ml浓度范围内具有良好的线性响应特性，且与平板活菌计数法有较好的一致性(r=0.92406，p＜0.001)，批内和批间误差(CV)＜15％。检测F1抗原的灵敏度为1ng／ml，其线性检测范围为1-300ng／ml。结论：UPT-ICA系统能快速特异地定性、定量检测鼠疫耶尔森氏菌F1抗原和全菌，在鼠疫的现场监测过程中具有广阔的应用前景。
[Abstract]:Objective : To develop an immunochromatography test strip based on Up - conversion Luminescence Technology ( UPT ) biosensor and immunochromatography assay ( ICA ) , and to develop an immunochromatography test strip based on up - conversion luminescent material ( UPT ) as a marker , which can be used to detect the high sensitivity and rapid field detection . Methods : The immune chromatography test strip was prepared by using the double antibody sandwich method as the marker and the double antibody sandwich method as the marker . The detection band signal ( V _ T ) and the quality control band signal ( V _ C ) were searched through the UPT biosensor , and the signal ratio ( V _ T / V _ C ) of the detection band and the control band was calculated . Results : The detection sensitivity of UPT - ICA test strip was 10 , 000CFU / ml , and good linearity was found in the concentration range of 10 ~ 4 ~ 10 ~ 8 CFU / ml ( r = 0.992 406 , p < 0.001 ) . The sensitivity of test F1 antigen was 1 ng / ml , and the linear detection range was 1 - 300ng / ml . Conclusion : The UPT - ICA system can be used for the rapid and specific qualitative and quantitative detection of the F1 antigen and the whole bacteria of the disease . The UPT - ICA system has a wide application prospect in the field monitoring of plague .

【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R378

【引证文献】