TRF (time-resolved fluorescence) immunochromatography reagent for rapidly and quantitatively detecting H-FABP (heart fatty acid-binding protein) and preparation method

Abstract

The invention discloses a TRF (time-resolved fluorescence) immunochromatography reagent for rapidly and quantitatively detecting H-FABP (heart fatty acid-binding protein) and a preparation method and belongs to the field of clinical medical diagnosis. The reagent comprises two parts including a test strip and a fluorescent liquid, wherein the test strip comprises a bottom plate, Fusion5, a nitrocellulose membrane and a water absorbent pad; the Fusion5, the nitrocellulose membrane and the water absorbent pad are horizontally and sequentially connected and fixed onto the bottom plate; the nitrocellulose membrane is coated with a detection line for H-FABP monoclonal antibodies 1 and a quality control line comprising rabbit IgG (immunoglobulin G) antibodies; the fluorescent liquid contains TRF microspheres labeled by H-FABP monoclonal antibodies 2 and TRF microspheres labeled by goat anti-rabbit antibodies. According to the reagent, the fluorescence intensity is improved by the aid of the TRF microspheres, background signals are reduced, meanwhile, the content of the H-FABP in whole blood, serum or plasma is quantitatively detected, and only 10-20 microliters of samples are required. The test strip is convenient, rapid, simple to operate, short in detection time, high in specialty, high in sensitivity, more accurate in detection result and applicable to rapid diagnosis for clinical POCT (point-of-care testing).

Description

Technical field [0001] The invention belongs to the field of clinical medical diagnosis, and specifically relates to a time-resolved fluorescence immunochromatographic reagent for rapid quantitative detection of H-FABP and a preparation method. Background technique [0002] Fatty acid binding proteins (FABPs) are abundant in active fatty acid metabolism tissues, such as the heart and liver, and their main function is to promote the transport of long-chain fatty acids in cells. Nine different types of FABP have been identified, of which H-FABP (heart-type fatty acid binding protein) is the most widespread because it is abundantly present in cardiomyocytes. The combination of its low molecular weight and cytoplasmic location makes H-FABP a highly sensitive early marker of acute coronary syndrome (especially within 6 hours of onset of chest pain), which can be detected 30 minutes after an ischemic attack. This may be because after myocardial ischemia and myocardial necrosis, it qui...

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.

[0022] 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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