Fluorescence immunochromatographic assay test card for testing cat serum amyloid A and preparation method

Abstract

The invention belongs to the field of animal epidemic disease detection and discloses a fluorescence immunochromatographic assay test card for testing cat serum amyloid A and a preparation method. Thefluorescence immunochromatographic assay test card is characterized by comprising a base plate, a sample pad, a combination pad, a nitrocellulose membrane and a water absorption pad are sequentiallyjointed on the base plate, the combination pad is provided with a fluorescent microsphere marked SAA monoclonal antibody and fluorescent microsphere marked goat anti-chicken IgY, The nitrocellulose membrane is provided an SAA monoclonal antibody coated test line T and a chicken IgY coated quality control line C. The fluorescence immunochromatographic assay test card for testing cat serum amyloid Ais high in sensitivity and reliable in test result.

Description

technical field [0001] The invention discloses an immunofluorescence chromatography detection card, specifically, an immunofluorescence chromatography detection card for detecting cat serum amyloid A, and the invention also discloses the immunofluorescence chromatography detection card Preparation. Background technique [0002] Serum amyloid A (SAA) is a major acute-phase protein present in many species, including humans, dogs, cats, and horses. Once the body is stimulated by inflammation caused by infection, trauma and surgery, the concentration of SAA will increase rapidly within a few hours; at the same time, the half-life of SAA is very short, and the concentration of SAA will decrease rapidly as the source of inflammation is removed. [0003] The gene sequence of SAA is highly conserved among vertebrates. Human SAA contains 104 amino acids. The cat's SAA is equivalent to an additional 111 amino acid sequences inserted in the middle region of the human SAA molecule. ...

AI Technical Summary

Problems solved by technology

[0005] The existing SAA detection method is mainly enzyme-linked immunoassay. Due to the long cycle of enzyme-linked immunoassay, it cannot be used on automatic biochemical analyzers, and it is not suitable for rapid detection in emergency. The turbidimetric method is simple and economical, and can be used on the automatic biochemical analyzers in most hospitals, especially in emergency departments, where rapid quantitative detection can be achieved. After the reaction, aggregated particles are formed, and at a certain wavelength, the content of the analyte in the sample can be measured by measuring the turbidity generated by the aggregate

[0006] Chinese patent ZL 201210575603.5 is a kit for rapid quantitative detection of serum amyloid A. Dry colloidal gold complex solution, washing solution and blocking solution; the reaction plate is a colloidal gold detection plate, and the nitrocellulose membrane on the reaction plate is coated with another purified SAA monoclonal antibody or polyantibody directed at another reaction determinant; However, this method uses the immunodiafiltration method, which needs to dissolve and dilute the anti-SAA gold standard solution freeze-dried product, dilute the sample, add blocking solution dropwise, infiltrate, drop the diluted anti-SAA gold standard solution dropwise, add washing solution, and finally detect, etc. A series of steps are very cumbersome and time-consuming, which is not conducive to early and rapid diagnosis and treatment of diseases

In addition, the patent uses colloidal gold as a tracer, and the immune marker relies on electrostatic adsorption, which has defects such as low sensitivity, difficulty in quantification, poor repeatability and stability, and cannot fully meet the clinical detection needs.

[0007] Chinese patent ZL201510497125.4 discloses a human serum amyloid A detection kit, the kit includes reagent R1, reagent R2, the reagent R1 is an appropriate buffer; the reagent R2 is human serum Amyloid A antibody sensitized polystyrene latex particles, placed in an appropriate buffer, but this method uses two buffers, the liquid needs to be refrigerated, and an automatic biochemical analyzer such as Hitachi 7080 is required for detection. It is relatively large, not suitable for on-site detection, and cannot be diagnosed and treated as early as possible

[0008] Chinese patent ZL201511020151.4 discloses a fluorescent immunochromatography detection kit for serum amyloid A, including a kit body and a diluent bottle, the kit body includes an immunochromatography detection card, and the immunochromatography detection The card includes a PVC liner and a sample pad fixed on the PVC liner, a probe fixing pad, a nitrocellulose membrane and absorbent paper, the sample pad is lapped on the probe fixing pad, and the probe fixing pad and the absorbent paper respectively lapped on both sides of the nitrocellulose membrane, the nitrocellulose membrane is provided with a detection line and a control line, the detection line is coated with serum amyloid A monoclonal antibody, and the control line is coated with There is goat anti-rabbit IgG; the probe immobilization pad is made by drying mixed fluorescent latex particles composed of serum amyloid A monoclonal antibody fluorescent latex particles and goat anti-rabbit IgG fluorescent latex particles; but this method has the following disadvantages: Fluorescent latex particles need to be coupled with albumin first, and then coupled with serum amyloid A monoclonal antibody. Although it can reduce the steric resistance of the double sandwich structure during the detection process, it also increases the coupling steps, which may Affect the repeatability of detection

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