| Fluorescence immunochromatography has become an indispensable quantitative detection method in clinical testing due to its advantages of high sensitivity,low cost and fast detection speed.Currently,a large number of clinical testing projects based on fluorescence immunochromatography have passed the country Approved by the Food and Drug Administration,and has entered practical clinical applications,such as inflammation,myocardium,thyroid function,hormones,drug screening and so on.As the key component of the analytical system fluorescence detection substance,its nature will play a vital role in the results of immunoassay.In this paper,a fluorescent nanospheres were prepared based on the 630nm/670nm fluorescence detection system,and two fluorescent dyes were prepared by synthesis.Cyanine dyes and fluorobilane Bodipy fluorescent dyes respectively.Cyanine dyes with fluorescence wavelength of650nm/670nm,which are used as water-soluble fluorescent dyes,form fluorescent antibodies through covalent binding and antibody coupling.The fluorine boron Bodipy fluorescent dye has a fluorescence wavelength of 630nm/650nm,BODIPY fluorescent dye is a kind of fluoroboron dipyrrole dye,which is extremely hydrophobic and soluble in organic solvent,Due to its extreme hydrophobicity,the dye and chloromethyl polystyrene microspheres are combined to form fluorescent microspheres by swelling method.The antibody was bound to the surface of fluorescent microspheres by fluorescence resonance energy transfer(FRET),and fluorescent microspheres emit630nm/670nm wavelength fluorescence consistent with the detection system.In this essay,the synthesized fluorescent microspheres were applied in the fluorescence immunochromatography platform.The cardiac troponin I(c Tn I)fluorescent immunofluorescent reagent was prepared by optimizing the application method and the parameters.The fluorescence immunochromatographic reagent prepared in this paper was in line with the linear correlation within the 0.01ng/ml-50ng/ml concentration range,the correlation coefficient R~2=0.9983,and the reagent was less than 10%at 0.3ng/ml and 1ng/ml concentrations.The reagent samples were collected at different concentrations for 7 days at 37 degrees centigrade,and the voltage signal value decreased by less than 15%.In terms of sensitivity,compared with commercialized commercial reagents,the lowest concentration of self-made reagent was 0.002ng/ml,while the minimum detection concentration of commercial reagents was only 0.008ng/ml,the sensitivity of the reagent has been significantly improved. |
