恙虫病东方体混合重组抗原应用于胶体金免疫层析试纸条的制备及效价检测  

Establishment of colloidal gold immunochromatographic assay to detect the antibody against Orientia tsutsugamushi

作　　者：赵芷若 周东明 操敏 陶清源 蔡旭燊 燕书豪 高豹 吕恒 谭伟龙 ZHAO Zhiruo;ZHOU Dongming;CAO Min;TAO Qingyuan;CAI Xusheng;YAN Shuhao;GAO Bao;L Heng;TAN Weilong(College of Food Science and Light Industry,Nanjing Tech University,Nanjing 211800,China;Center for Disease Control and Prevention of Eastern Theater Command,Nanjing 210002,China;National Light Industry Food Quality Supervision and Testing Nanjing Station,Nanjing 211800,China)

机构地区：[1]南京工业大学食品与轻工学院,江苏南京211800 [2]东部战区疾病预防控制中心,江苏南京210002 [3]国家轻工业食品质量监督检测南京站(南京工业大学),江苏南京211800

出　　处：《生物加工过程》2024年第2期196-203,共8页Chinese Journal of Bioprocess Engineering

基　　金：国家自然科学基金面上项目(81472932);江苏省自然科学基金(BK20170998);国家科技重大专项子课题(2018ZX10713003、2017ZX10303401);军队后勤重大项目(AWS16J020)。

摘　　要：胶体金免疫层析试纸条具有携带方便、操作简单且敏感性、特异性高等特点,近年来开始广泛应用于疾病的诊断。为了实现对恙虫病的快速检测,笔者利用双抗原夹心法,以胶体金标记恙虫病东方体56 kDa混合重组蛋白,56 kDa混合重组蛋白和兔抗恙虫病东方体多克隆抗体分别喷在检测线及质控线上,制备了用于检测恙虫病东方体抗体的胶体金免疫层析试纸条。本研究制备的胶体金试纸条检测限为61 ng/mL,和疟疾、伤寒、血吸虫、出血热以及斑点热等阳性血清无交叉反应,特异性达90%(54/60),检测的敏感性为91.96%(103/112),在4℃下可稳定保存6个月。本试纸条能够应用于人体血清中恙虫病东方体总抗体的检测。In recent years,colloidal gold immunochromatography strips were widely utilized in disease diagnosis,due to the convenience for carrying,simple operation,as well as high sensitivity and specificity.Herein,we developed a convenient approach for the rapid detection of Scrub typhus,using colloidal gold immunochromatographic strip to monitor the total antibody against Orientia tsutsugamushi.The double-antigen sandwich method was used in this case to spray the 56 kDa mixed recombinant protein of Orientia tsutsugamushi with gold onto the detection line,and the 56 kDa mixed recombinant protein and rabbit polyclonal antibody against Orientia tsutsugamushi onto the quality control line,respectively.As a result,our colloidal gold assay showed the detection limit of 61 ng/mL,and demonstrated no cross reactions with the positive sera of malaria,typhoid,schistosomiasis,haemorrhagic fever,spotted fever,etc.Moreover,the test strips proved to be stable at 4℃for 6 months,and this method could provide excellent specificity of 90%(54/60)and the detection sensitivity of 91.96%(103/112).Therefore,our colloidal gold immunochromatography strip can be applied to detect total antibody against Orientia tsutsugamushi in human serum.

关 键 词：恙虫病东方体 胶体金 混合重组蛋白 快速检测试纸条 

分 类 号：R513.2[医药卫生—内科学] Q599[医药卫生—临床医学]