Time-resolved fluorescence immunochromatographic test strip and kit for detection of cTnI, and preparation methods thereof

Abstract

The invention discloses a time-resolved fluorescence immunochromatographic test strip and kit for detection of cTnI, and preparation methods thereof. The test strip comprises a bottom liner and further comprises a sample pad, a conjugate pad, a coating membrane and absorbent paper successively arranged on the bottom liner, wherein the conjugate pad is coated with a cTnI monoclonal detection antibody labeled by fluorescence microspheres; the coating membrane comprises a test zone and a control zone arranged in parallel and spaced apart from each other; the test zone is coated with a cTnI monoclonal capture antibody capable of recognizing a single epitope, and the control zone is coated with a goat anti-mouse IgG antibody; and the coating membrane a nitrocellulose membrane bonded with a polymer, and the polymer is a material having light transmittance of 10% or below under irradiation of light with wavelengths of less than 450 nm and light transmittance of 95% or more under irradiation of light with wavelengths of 500 nm or more. The test strip of the invention can realize quick and quantitative detection, and is accurate and reliable in test results and high in sensitivity; the preparation method is simple in process and suitable for large-scale production; and the test strip has positive significance to quantitative detection of cTnI.

Description

technical field [0001] The invention belongs to the technical field of medical testing, in particular, the invention relates to a time-resolved fluorescent immunochromatographic test strip, a kit and a preparation method thereof for quantitative detection of cTnI. Background technique [0002] Troponin is a regulatory protein of muscle contraction. Cardiac troponin (cTn) is composed of subunits of three different genes: cardiac troponin T (cTnT), cardiac troponin I (cTnI), and troponin C (TnC). Currently, cTnT and cTnI are used for laboratory diagnosis of ACS. Because cTnI and the heteroplasmic body in skeletal muscle are encoded by different genes, have different amino acid sequences, and have unique antigenicity, its specificity is obviously better than that of CK-MB isozyme. When muscle tissue other than myocardium is damaged or diseased, CK and CK-MB may increase, but cTnI will not exceed its critical value. Because their content in normal serum is very small, they in...

AI Technical Summary

Problems solved by technology

[0005] 2. Gold standard method - this method is fast, simple and easy to observe, but the sensitivity is not high

[0006] 3. Transmission immunoturbidimetry——This method is simple, fast, and can be automated, and is suitable for batch detection. However, the methodology and clinical application of immunoturbidimetry still need further verification

Various test strips and instruments based on immunochromatography, chromatography and dry chemistry techniques will affect the activity of microproteins in the matrix due to differences in temperature, humidity and pH, thereby affecting the results

The methodological flaws of some POCT instruments, such as poor sensitivity and repeatability, and narrow linear range, are only used for reference in emergencies or emergencies, and need to be sent to the laboratory for re-examination if necessary.

[0012] The current immunofluorescence chromatography device uses the reflection method to detect the fluorescent signal on the porous membrane. The fluorescence detector captures the specific antibody modified by the fluorescent dye on the surface of the porous membrane, but cannot detect the fluorescent signal inside the porous membrane, which leads to the detection of Sensitivity drop