Time-resolved fluorescent immunochromatographic test strip for detecting vancomycin as well as preparation method and application thereof
Some embodiments of the disclosure provide a time-resolved fluorescent immunochromato-graphic test strip for detecting vancomycin as well as a preparation method and application thereof. In some embodiments, the test strip includes a bottom plate and a sample absorption pad. A fluorescent microsphere pad, a nitrocellulose membrane coated with a vancomycin-carrier protein conjugate, and an absorbent pad are sequentially overlapped and pasted on the bottom plate. The fluorescent microsphere pad is sprayed with a fluorescent microsphere-labeled vancomycin monoclonal antibody, and the vancomycin monoclonal antibody is prepared by using a vancomycin-bovine serum albumin conjugate as an immunogen.
1. A test strip-based detection method of vancomycin, comprising the steps of:
(1) adding pure vancomycin of different concentrations to a human serum matrix excluding vancomycin to prepare a calibrator, concentrations of vancomycin in the calibrator being 40 μg/mL, 20 μg/mL, 10 μg/mL, 5 μg/mL, and 0 μg/mL, sequentially;
(2) diluting the calibrator 100-1,000 times and loading the diluted calibrator to a sample absorption pad of a test strip for immunochromatography;
(3) determining a T/C ratio of a time-resolved fluorescent intensity of a detection area to the time-resolved fluorescent intensity of a quality control area to obtain a function relation of the concentration of the calibrator and the T/C ratio;
(4) taking a sample to be tested and loading the sample to the sample absorption pad of the test strip for immunochromatography;
(5) determining a ratio of the time-resolved fluorescent intensity of the detection area to the time-resolved fluorescent intensity of the quality control area; and
(6) calculating a content of vancomycin in the sample to be tested according to the function relation.
2. The method according to claim 1 wherein the sample to be tested is loaded in an amount of 1-100 μL.