Preparation and application of fluorescence immunochromatography test strip for exosome quantification

Abstract

The invention discloses preparation and application of a fluorescence immunochromatography test strip for exosome quantification, and particularly relates to the field of biological detection. The test strip comprises a back plate, a sample pad, a combination pad, an NC membrane and a water absorption pad, wherein the sample pad, the combination pad, the NC membrane and the water absorption pad are pasted on the back plate, the combination pad is in lap joint with one end of the sample pad and one end of the NC membrane, the other end of the NC membrane is in lap joint with the water absorption pad, and a detection line and a quality control line are sprayed on the NC membrane. A mouse anti-human CD9 antibody marked by fluorescent microspheres and rabbit IgG marked by fluorescent microspheres are attached to the combination pad, a mouse anti-human CD9 monoclonal antibody is coated on the detection line, and a goat anti-rabbit antibody is coated on the quality control line. By utilizing the optimized fluorescence immunochromatography technology, the concentration of the exosome can be measured more conveniently, quickly and accurately, and a new technical means is provided for the detection of the concentration of the exosome.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to the preparation and application of a fluorescent immunochromatographic test strip for quantifying exosomes. Background technique [0002] Exosomes are small vesicles with a phospholipid bilayer structure secreted by living cells, with a diameter of 30-150nm and a density of 1.13-1.19g / ml, which can exist in various body fluids, such as serum, plasma, saliva, urine fluid, ascites, spinal fluid, milk, etc. Exosomes contain a variety of biomolecules, such as mRNA, miRNA, proteins, lipids, etc., which can be delivered to recipient cells, thereby changing the physiological or pathological functions of recipient cells. In recent years, exosomes have attracted widespread attention as intercellular information transfer tools and biomarkers of various diseases. Exosomes have great potential in the fields of biomedicine and disease diagnosis. [0003] The detection and quant...

AI Technical Summary

Problems solved by technology

[0003] The detection and quantification of exosomes is a routine experiment in exosome research. At present, there are many detection and quantification techniques that can be used for exosomes, such as dynamic light scattering technology, nanoparticle size tracer analysis technology, and adjustable resistance pulse transmission. Sensitive technology, flow cytometry and nano flow technology are often used to measure the concentration of exosomes, but these technologies also have certain limitations, among which dynamic light scattering technology, nanoparticle size tracer analysis technology, Adjustable resistance pulse sensing technology cannot remove the pollution of non-exosome particles in complex samples, resulting in unrealistic exosome concentration data. The detection sensitivity of flow cytometry is about 500nm, which cannot directly detect exosomes. Immunomagnetic beads can only be used as carriers to perform immunocapture of exosomes, and then carry out concentration detection. Because this process is too cumbersome, the operation is difficult, and the failure rate of the experiment is extremely high. Although nano flow technology can directly detect exosomes in samples The concentration of exosomes, but this instrument is too expensive, and ordinary laboratories do not have this instrument, resulting in the inability to quickly obtain concentration data for exosome samples to be tested, which brings inconvenience to downstream experiments

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