Fluorescence counting system for quantifying viruses or antibodies on an immobilized metal substrate by using an antigen-antibody reaction

Abstract

The present invention relates to a system capable of performing simple and rapid inspection of an antigen equivalent to the immune chromatographic method with accuracy good as a PCR method. An embodiment relates to a novel fluorescence counting system for quantifying viruses or antibodies in an analyte which comprises an unit of providing an antigen or antibody phase solidified substrate by an aggregation method with quantum crystals, an unit for making a labeling liquor and labeling a virus or an antibody to be measured in the analyte by an antigen-antibody method, an unit of exciting the fluorescently labeled virus or antibody by a surface plasmon excitation method, and an unit of counting fluorescent points in an excited fluorescent screen to quantify the virus or antibody in the analyte.

1 . A measuring metal substrate used for fluorescence labelling immune assay comprising (a) a metal substrate and (b) an antibody or antigen solidified phase configured to capture a first for capturing an antigen or a first antibody respectively as a target in a sample added made on the a metal substrate using (i) a plasmon metal complex solution comprising plasmon metal complex quantum crystals and (ii) a buffer solution comprising the first antibody or the antigen;
 wherein there is an electrode potential difference between the metal substrate and the plasmon metal complex quantum crystals;   wherein the plasmon metal complex quantum crystals have (a) a size in a range of 50 nm to 150 nm and are (b) substantially free of agglomeration of coagulated plasmon metal quantum crystals with each other,   wherein the first antibody or the antigen from the plasmon metal complex solution is immobilized on the metal substrate with the plasmon metal complex quantum crystals that are coagulated on the metal substrate, to form an immobilized first antibody or an immobilized antigen respectively;   wherein the measuring substrate is configured for a quantitative detection of a target as a fluorescent spot via fluorescence labelling immune assay using an antigen-antibody immune reaction; and   wherein the measuring substrate possess a plasmon enhancing ability for exciting the target to be labelled with a fluorescent material by irradiation of an exciting light for detecting the target antigen as the fluorescent spot through a fluorescent image.   
     
     
         2 . The measuring substrate of  claim 1 , wherein pH of the buffer solution is about 7 or more. 
     
     
         3 . The measuring substrate of  claim 1 , wherein the plasmon metal complex solution comprises a density of the plasmon metal quantum crystals in a range of 1000 to 5000 ppm. 
     
     
         4 . A system comprising:
 a) the measuring substrate as claimed in  claim 1 ,   b) a fluorescence labelling unit comprising a labelling fluorescence material configured to label the target antigen to form a labelled target by an antigen-antibody immune reaction;   c) a fluorescence imaging unit configured to make a fluorescence image of the labelled target trapped on the first antibody or antigen solidified phase as the fluorescent spot using the excitation light, and   d) a counting unit for counting fluorescence spots and quantify the target antigen;   wherein the target antigen is an outside protein of a virus, or the target antigen is the virus itself and wherein the target is not an entity present inside the virus and separate from the virus; and   wherein the system is configured to perform a fluorescence labelling immune assay.   
     
     
         5 . The measuring substrate of  claim 4 , wherein the target antigen is the virus. 
     
     
         6 . The measuring substrate of  claim 5 , wherein the virus is an enveloped virus. 
     
     
         7 . The measuring substrate of  claim 6 , wherein the enveloped virus comprises influenza and/or COVID-19 virus. 
     
     
         8 . The system of  claim 4 , further comprising a device to blow air to stop agglomeration of the coagulated plasmon metal quantum crystals on the metal substrate. 
     
     
         9 . The system of  claim 4 , wherein the fluorescence labeling unit is dropped on the metal substrate to form the labelled target trapped on the first antibody. 
     
     
         10 . The system of  claim 4 , wherein the fluorescence imaging unit comprises a light source to irradiate the excitation light having a wavelength range suitable for fluorescence for the labelled target attached with the coagulated plasmon metal complex. 
     
     
         11 . The system of  claim 4 , wherein the counting unit is configured to binarize the fluorescence image to adopt the fluorescence spot and quantitatively count the fluorescence spot. 
     
     
         12 . The system of  claim 4 , wherein the fluorescence image is binarized with an analysis condition comprising one or more of a brightness, an area, and a circularity of the fluorescence spot in the fluorescence image. 
     
     
         13 . The system of  claim 4 , wherein the target antigen is labelled by a sandwich method of the antigen-antibody immune reaction. 
     
     
         14 . The system of  claim 4 , wherein the system is configured to detect more than one type of the target antigen. 
     
     
         15 . The system of  claim 4 , target antigen comprising influenza and/or COVID-19 virus. 
     
     
         16 . The system of  claim 4 , wherein the system comprises a filter for labeling of the fluorescence material having different wave range depending on the target antigen. 
     
     
         17 - 30 . (canceled)