中文摘要：

目的研制基于上转发光免疫层析技术（UPT-LF）检测鼠疫耶尔森菌（简称鼠疫菌）、炭疽杆菌芽孢、布鲁菌的试纸，与BioThreatAlert免疫层析（BTA）试纸的检测和操作性能进行对比。方法以上转发光纳米颗粒作为生物示踪物，采用双抗体夹心模式分别研制检测鼠疫菌、炭疽杆菌芽孢、布鲁菌的uPT．LF试纸。分别配制10^10、10^9、10^8、10^7、10^6、10^5、0CFU／ml系列浓度的鼠疫菌、炭疽杆菌芽孢、布鲁菌标准品以及10^9CFU／ml系列浓度的27种特异性评价菌株菌液，采用UPT-LF试纸对其进行检测，对3种UPT-LF试纸的敏感性、精密性、线性和特异性进行分析评价。配制模拟阳性样品，分别采用uPT-LF和BTA试纸对10^7、10^6、10^5、0CFU／ml系列浓度的标准品以及模拟阳性样品进行检测，比较UPT-LF和BTA试纸的检测时间、敏感性以及鼠疫菌、炭疽杆菌芽孢、布鲁菌的检出率。结果鼠疫菌UPT—LF、炭疽杆菌芽孢uPrr—LF、布鲁菌UPT-LF试纸的敏感性均达到10^5CFU／ml，各系列浓度检测带信号／质控带信号（T／C）值的变异系数均≤15％，lg[T／C值-临界值（cut—off值）]与lg（标准品浓度）的相关系数分别为0．996、0．998、0．999（F=1647．57、743．51、1822．17，P值均〈0．001）。鼠疫菌UPT-LF和布鲁菌UPT-LF试纸检测特异性评价菌株时，均无非特异性交叉反应，炭疽杆菌芽孢UPT—LF试纸检测枯草芽孢杆菌分离株2#芽孢、蜡样芽孢杆菌分离株1#芽孢存在非特异交叉反应，检测其他特异性评价菌株均无非特异性交叉反应。UPT-LF试纸检测鼠疫菌、炭疽杆菌芽孢、布鲁菌样品的时间分别为33、36、37min，BTA为115、115、111min；UPT-LF和BTA试纸检测0CFU／ml标准品的阴性率均为5／5；UPT-LF试纸检测炭疽杆菌芽孢、鼠疫菌、布鲁菌的敏感性均达到10^5CFU／ml，BTA试纸分别为10^6、10^6、10^5CFU／ml；模拟样品检测中，UP

英文摘要：

Objective To develop an up-converting phosphor technology based lateral flow （UPT- LF） assay for rapid and quantitative detection of Yersinia pestis, Bacillus anthracis spore and Brucella spp. and make the comparison with BioThreat Alert （BTA）test strips（ Tetracore Inc. , USA）. Methods Using up-converting phosphor nano-particles （UCP-NPs） as the bio-marker, three double-antibody-sandwich model based UPT-LF strips including Plague-UPT-LF, Anthrax-UPT-LF, Brucella-UPT-LF were prepared and its sensitivity, accuracy, linearity and specificity were determined by detecting 10^ 10, 10^9, 10^8 , 10^7, 10^6, 10^5 and 0 CFU/ml series of concentrations of Y. pestis, B. anthracis, Brucella standards and other 27 kinds of 109 CFU/ml series of contrations of bacteria strains. Furthermore, the speed, sensitivity and accuracy of bacteria standards and simulated sample detection were compared between UPT-LF and BTA system. Results The detection limit of Plague-UPT-LF, Anthrax-UPT-LF and Brucella-LF was 105 CFU/ml. The CV of series of bacteria concentrations was ≤ 15%, and the r between lg（T/C -cut-off）and lg （concentration） was 0. 996,0. 998 and 0. 999（F values were 1 647.57,743.51 and 1 822. 17. All the P values were 〈0. 001） , respectively. The specificity of Plague-UPT-LF and Brueella-LF were excellent, while that of Anthrax-UPT-LF was a little bit regretful because of non-specific reaction with two isolates of B. subtilis and one B. cereus. On- site evaluation showed the detection time of UPT-LF for all Y. pestis ,B. anthracis spore and Brucella spp. was 33,36 and 37 min,while BTA was 115,115 and 111 min,which revealed the higher detection speed and sensitivity of UPT-LF comparing with BTA. The negative rate of two methods for blank standard was both 5/ 5, the sensitivity of UPT-LF for Y. pestis,B, anthracis spore and Brucella spp. was all 105 CFU/ml,then BTA was 10^6 , 10^6 and 10^5 CFU/nd, respectively. The detection rate of UPT-LF for all three bacteria analog positive samples was 16/16, whi