用于新型冠状病毒快速诊断的荧光免疫层析试纸条的制备及其细胞成像  

Preparation of two-photon fluorescence immunochromatographic strip for rapid diagnosis of SARS-CoV-2 and cell-imaging

作　　者：黄池宝 杨斌[2,3] 康帅[1] 杨丽 吕国岭[1] 余福勋 HUANG Chibao;YANG Bin;KANG Shuai;YANG Li;LYU Guoling;YU Fuxun(School of Information Engineering,Zunyi Normal University,Zunyi 563002,Guizhou,China;Department of Central Laboratory,Guizhou Provincial People's Hospital,Guiyang 550002,Guizhou,China;NHC Key Laboratory of Pulmonary Immunological Diseases,Guizhou Provincial People's Hospital,Guiyang 550002,Guizhou,China;Henry Fok School of Biology and Agriculture,Shaoguan University,Shaoguan 512005,Guangdong,China)

机构地区：[1]遵义师范学院信息工程学院,贵州遵义563002 [2]贵州省人民医院中心实验室,贵州贵阳550002 [3]贵州省人民医院卫健委肺脏免疫性疾病重点实验室,贵州贵阳550002 [4]韶关大学英东生物与农业学院,广东韶关512005

出　　处：《精细化工》2021年第7期1367-1373,共7页Fine Chemicals

基　　金：贵州省高层次创新型人才培养计划项目——“百”层次人才([2016]5683);贵州省科技支撑计划项目([2020]4Y001,[2020]4Y004,[2018]2785);贵州省教育厅自然科学研究项目(KY[2019]068,KY[2018]028);贵州省教育厅自然科学重点研究项目(黔教合KY字[2014]296);国家自然科学基金(21562050,[2019]81960001);贵州省第八批科技创新人才团队建设专项基金[(2015)4007];贵州省科学技术基金项目(黔科合J字[2015]2146)。

摘　　要：合成了双光子荧光标记探针LP并对其结构进行了表征,通过LP标记SARS-CoV-2的N-蛋白抗原(Ag)获得LP-Ag,并将LP-Ag制成免疫层析试纸条(TPICS)/试剂盒(TPICK)。临床应用研究表明,与核酸法(PCR)/胶体金法比较,TPICS的阳性符合率、阴性符合率和总符合率分别为98.99%/98.10%、99.55%/99.53%和99.38%/99.06%,其一致性系数Kappa值(K)为0.972/0.975,灵敏度、特异性和准确度分别为98.99%/98.10%、99.55%/99.53%和99.38%/99.06%。检测试剂与参比试剂检测结果一致(K>0.75),无统计学差异。TPICS/TPICK检测所用稀释血样一般为25~50μL(合全血样5~10μL),检测时间在10 min内,既避免了核酸法操作繁琐与检测周期过长的缺点,又克服了胶体金法因吸附官能团脱落而导致的不稳定性,具有良好的应用价值。LP-Ag可用于细胞中SARS-Co V-2成像以鉴定其阴阳性。A two-photon fluorescence labelling probe LP was synthesized and characterized.LP-Ag was obtained by labelling the N protein antigen(Ag)of SARS-CoV-2 with LP,and then the LP-Ag was made into an immunochromatographic test strip(TPICS)/kit(TPICK).Clinical application results showed that compared with those of nucleic acid method(PCR)/colloidal gold method,the positive coincidence rate,negative coincidence rate and total coincidence rate of TPICS were 98.99%/98.10%,99.55%/99.53%and 99.38%/99.06%,respectively.The consistency coefficient Kappa value(K)was 0.972/0.975.The sensitivity,specificity and accuracy were 98.99%/98.10%,99.55%/99.53%and 99.38%/99.06%,respectively.The results of detection reagent and reference reagents were consistent(K>0.75)without statistical difference.The dilute blood sample used in TPICS/TPICK detection was generally 25~50μL(whole blood sample 5~10μL),and the test time was within 10 min.This method avoided the disadvantages of complex operation and long detection period of nucleic acid method,and overcame the instability caused by the fall off of the adsorption functional group of colloidal gold method.Therefore,TPICS/TPICK showed a superior application prospect.LP-Ag could be used for SARS-CoV-2 imaging in cells to identify negative and positive.

关 键 词：COVID-19/SARS-CoV-2 N-蛋白 免疫层析试纸条 双光子荧光 试剂盒 功能材料 

分 类 号：R563.1[医药卫生—呼吸系统] O657.3[医药卫生—内科学]