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

The Preparation of Two-photon Fluorescence Immunochromatographic Strip for Rapid Diagnosis of SARS-COV-2 and Cell-imaging

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（PR）、阴性符合率（NR）和总符合率（TR）分别为98.73 % / 98.26 %、98.76 % / 99.20 %和98.75 % / 98.75 %，其一致性系数kappa值（K）为0.972 / 0.975，灵敏度（SE）、特异性（SP）和准确度（AC）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。检测所用稀释血样一般为25~50 μL（合全血样5~10 μL），检测周期10 min内。它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-COV-2成像以鉴定阴阳性。

Abstract:

A two-photon fluorescence labelling probe LP was synthesized and characterized, and LP-Ag was obtained by LP labelling N-protein antigen (Ag) of SARS-COV-2, and LP-Ag was made into an immunochromatographic test strip (TPICS)/ kit (TPICK). Clinical applications showed that, in comparison with the nucleic acid method (PCR)/ the colloidal gold method, of TPICS positive coincidence rate (PR), negative coincidence rate (NR) and total coincidence rate (TR) were 98.73 % / 98.26 %, 98.76 % / 99.20 % and 98.75 % / 98.75 %, and the consistency coefficient kappa value (K) was 0.972 / 0.975, and sensitivity (SE), specificity (SP) and accuracy (AC) were 98.99 % / 98.10 %, 99.55 % / 99.53 % and 99.38 % / 99.06 %, respectively. Detection results of Detection reagent and reference reagents were consistent (K > 0.75) with no statistical difference. Dilute blood sample used for testing is generally 25~50 μL (Whole blood sample 5~10 μL), and the test period is within 10 min. It not only avoids the disadvantages of cumbersome nucleic acid operation and long detection period, but also overcomes the instability of colloidal gold due to the shedding of the adsorbed functional groups, and has good application value. LP-Ag can be used for SARS-COV-2 imaging in cells to identify negative and positive.