Time-resolved fluorescence immunochromatography reagent used for rapid quantitative detection of CK-MB, and preparation method thereof

Abstract

The invention discloses a time-resolved fluorescence immunochromatography reagent used for rapid quantitative detection of CK-MB, and a preparation method thereof, and belongs to the field of clinical diagnose. The time-resolved fluorescence immunochromatography reagent is composed of a test paper strip and a fluorescent liquid; the test paper strip comprises a base plate, Fusion5, a nitrocellulose membrane, and a water absorption pad; Fusion5, the nitrocellulose membrane, and the water absorption pad are connected in sequence in the horizontal direction, and are fixedly arranged on the base plate; the nitrocellulose membrane is coated with a CK-MB monoclonal antibody 1 detection line and a quality control line composed of rabbit IgG antibodies; and the fluorescent liquid comprises CK-MB monoclonal antibody 2 labelled fluorescent microspheres, and goat anti-rabbit antibody labeled fluorescent microspheres. According to the time-resolved fluorescence immunochromatography reagent, time-resolved fluorescent microspheres are used for increasing fluorescence intensity, reducing background signals, and realizing quantitative detection of CK-MB content of whole blood, serum, or blood plasma at the same time; and only 10 to 20ml of a sample is needed. Application of the time-resolved fluorescence immunochromatography reagent is convenient and rapid; operation is simple; detection time is short; specificity and sensitivity are high; detection results are more accurate; and the time-resolved fluorescence immunochromatography reagent is suitable for clinical POCT rapid diagnosis.

Description

technical field [0001] The invention belongs to the field of clinical medical diagnosis, and in particular relates to a time-resolved fluorescence immunochromatography reagent for rapid and quantitative detection of CK-MB and a preparation method thereof. Background technique [0002] CK-MB is one of the three isoforms of creatine kinase (CK) (the other two are CK-BB and CK-MMOL / L). CK is a main enzyme of muscle metabolism. CK-MB consists of two subunits (MW=40000 each): subunit M for muscle and subunit B for brain. CK-MB mainly exists in the myocardium, and a small amount exists in the skeletal muscle. Quantitative determination of CK-MB is part of routine cardiac examination and is helpful for the diagnosis of AMI. When AMI occurs, CK-MB appears in the blood circulation and indicates that the myocardium has been damaged. CK-MB concentrations rise within 4-8 hours of pain onset, peak at 12-24 hours, and then decline to normal levels after 48 hours. CK-MB is the marker ...

AI Technical Summary

Problems solved by technology

Enzyme-linked immunosorbent assay is complex in operation, long in detection time, and narrow in detection range; latex-enhanced immunoturbidimetry needs to be combined with large-scale instruments, and the cost is high; colloidal gold immunochromatography for qualitative or quantitative detection has low sensitivity and narrow linear range, which cannot meet quantitative , Accurate detection requirements; Most of the fluorescent immunochromatography methods use fluorescein or other substances as light sources. Although they have a high linear range, they cannot guarantee low-end sensitivity due to background signal interference.

[0006] Although time-resolved fluorescent immunochromatography has been widely used in the detection of various items, most of them use time-resolved fluorescent microspheres immobilized on the conjugate release pad, and the imprecision is large due to the uneven release of the microspheres. , can’t meet the items with high sensitivity requirements, especially the detection of troponin I

In addition, in order to detect whole blood samples, the sample pads of most manufacturers use artificially treated glass fibers as conjugate release pads, which also increases the imprecision

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