CN112345773A - Immunoassay device for in vitro instant determination of ABO and/or Rh blood type of human - Google Patents
Immunoassay device for in vitro instant determination of ABO and/or Rh blood type of human Download PDFInfo
- Publication number
- CN112345773A CN112345773A CN202011015285.8A CN202011015285A CN112345773A CN 112345773 A CN112345773 A CN 112345773A CN 202011015285 A CN202011015285 A CN 202011015285A CN 112345773 A CN112345773 A CN 112345773A
- Authority
- CN
- China
- Prior art keywords
- quality control
- reaction
- internal standard
- control internal
- immunoassay device
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/80—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood groups or blood types or red blood cells
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
Abstract
The invention discloses an immunoassay device for in vitro real-time determination of ABO and/or Rh blood type of a human, which comprises an upper cover, a reaction membrane, an absorption liner, a back lining and a base. The upper cover is provided with a plurality of sample holes, a reaction film is respectively arranged below each sample hole, the reaction films are respectively coated with antibodies for determining blood type or positive quality control internal standard substances and negative quality control internal standard substances for monitoring reaction effectiveness, the absorption liner and the reaction films are arranged side by side at intervals so that the reaction films below adjacent holes are spaced, the back lining is positioned on the opposite side of the upper cover relative to the reaction films and the absorption liner, one surface of the back lining is attached to the reaction films and the absorption liner, and the other surface of the back lining is in contact with the base and is supported and fixed by the base. The immunoassay device for in vitro real-time determination of the ABO and/or Rh blood type of the human body is provided with the quality control internal standard, can be used for various situations such as hospital, family and field emergency, and can quickly, accurately and simply detect the ABO and/or Rh blood type of the human body.
Description
Technical Field
The invention relates to the field of medical blood type typing detection, in particular to an immunoassay device for in vitro real-time determination of ABO and/or Rh blood type of a human body.
Background
The techniques for typing human blood types fall into 2 general categories, genotyping for DNA and serotyping for antigen/antibody. Most of the current blood typing detection methods used in clinical examination are only suitable for central laboratories. The problems of low detection speed, inconvenient operation, high cost and the like cannot be avoided. In recent years, with the implementation of medical resource sinking and classified diagnosis and treatment, a simple, convenient, rapid and accurate blood type typing detection technology is urgently needed.
In view of the above, many people have conducted some research and proposed some technical solutions, but the following problems generally exist in the technical solutions: fast and inaccurate, lack of quality control, inconvenient operation, difficult production and manufacture and high cost.
For example: the Chinese patent with publication number 2013025763 discloses a blood grouping card for ABO and Rh-D. The card uses plastic or gummy paper as a substrate, and dried standard anti-A, anti-B and anti-D serum is coated in a serum groove on the substrate, so as to carry out blood grouping. The identification card is easy to carry, has good applicability and easy storage, and can make up for the defects of condensation collection in the traditional slide method. The method lacks quality control, and false negative and false positive are easy to occur.
The Chinese patent with publication number 201382943 discloses a reagent card for detecting ABO and RhD blood typing of human by shift chromatography, wherein an anti-A detection line, an anti-B detection line, an anti-RhD detection line and a quality control line are arranged on a reaction membrane. The detection line is a corresponding monoclonal antibody, and the quality control line is a polyclonal antibody. And displaying red by using the detection line and the quality control line to perform corresponding result judgment. The method has low detection cost. However, the antigen and antibody need secondary reaction at the quality control line, which increases the detection time. Meanwhile, because the blood color is red, the detection line and the quality control line carry out result interpretation according to the displayed red color, the error is larger, and the specificity needs to be improved.
The Chinese patent application with the application number of 201811096877.X discloses a reagent card for detecting ABO and RhD blood typing of human by adopting a percolation method, but the reagent card is lack of quality control and is easy to have false positive and false negative. The process needs to be kept still for reaction for a period of time, and the instant detection cannot be carried out.
Disclosure of Invention
The invention aims to provide an immunoassay device for in vitro real-time determination of ABO and/or Rh blood type of a human body, which can simply, quickly and accurately determine the ABO and/or Rh blood type of the human body.
The invention provides an immunoassay device for in vitro real-time determination of ABO and/or Rh blood type of a human body, which comprises an upper cover, a reaction film, an absorption liner, a backing and a base, wherein the upper cover is provided with a plurality of sample holes, the reaction film is respectively arranged below each sample hole, the reaction film below each sample hole is respectively coated with an antibody for determining the blood type or a positive quality control internal standard substance and a negative quality control internal standard substance for monitoring the reaction effectiveness, the absorption liner and the reaction film are arranged side by side at intervals, so that the reaction films below adjacent sample holes are spaced, the backing is positioned on the opposite side of the upper cover relative to the reaction films and the absorption liner, one surface of the backing is attached to the reaction films and the absorption liner, and the other surface of the backing is in contact with the base and is supported and fixed by the base, the base and the upper cover are assembled together in a manner of stable fit.
Preferably, the plurality of sample wells comprise a measurement well, a positive quality control internal standard well and a negative quality control internal standard well, the positive quality control internal standard is an anti-human RBC antibody, and the negative quality control internal standard well is an anti-goat IgG antibody.
Preferably, the kit comprises three measuring holes, and the corresponding reaction membranes below the three measuring holes are coated with an anti-A antibody, an anti-B antibody and an antibody for identifying Rh blood type respectively.
Preferably, three of the assay wells are arranged in a row, and the positive quality control internal standard well and the negative quality control internal standard well are arranged in a row.
Preferably, the measuring hole has an upper port and a lower port which are circular, and the diameter of the upper port is larger than that of the lower port, and the edge of the upper port and the edge of the lower port are connected by a gentle continuous surface.
Preferably, the internal control standard hole and the internal control standard hole both have a circular upper port and a square lower port, the projection of the upper port vertically facing the lower port can completely cover the lower port, and the edge of the upper port is connected with the edge of the lower port through a smooth continuous surface.
Preferably, the anti-a antibody and the anti-B antibody are respectively selected from any one of an IgM antibody, an IgG antibody or a mixture of the IgM and IgG antibodies, and the antibody for identifying the Rh blood group is selected from any one of an anti-D antibody, an anti-C antibody and an anti-e antibody.
Preferably, the substrate of the reaction membrane is selected from any one of cotton paper, printing paper, blotting paper, nitrocellulose membrane and cellulose acetate membrane or any combination thereof.
Preferably, the absorbent pad is selected from one or any combination of filter paper, cellulose fiber, and tissue paper.
Preferably, the edge of the upper cover is provided with a protrusion, the edge of the base is provided with a groove, and the upper cover and the base can be assembled together through the firm fit of the protrusion and the groove.
The immunoassay device for in vitro real-time determination of the ABO blood type and/or the Rh blood type of the human body can be used in hospitals, families, field emergencies and other situations due to the self-contained quality control internal standard for monitoring the effectiveness of the detection reaction, can quickly, accurately and simply detect the ABO blood type and/or the Rh blood type of the human body without additional instruments or manpower, improves the detection efficiency and reduces the detection cost.
Drawings
FIG. 1 is a schematic view of the external structure of the immunoassay device for in vitro real-time determination of ABO and/or Rh blood type of a human body.
FIG. 2 is a schematic view showing the structure of an upper cover of the immunoassay device in FIG. 1.
FIG. 3 is a schematic view of the structure of the reaction membrane, absorbent pad and backing of the immunoassay device of FIG. 1.
FIG. 4 is a schematic view of the structure of the base of the immunoassay device of FIG. 1.
Fig. 5 is a diagram exemplarily showing a reaction principle for determining a blood type using the immunoassay device of fig. 1.
Fig. 6 is a diagram exemplarily showing result determination in blood type determination using the immunoassay device of fig. 1.
Detailed Description
The invention will be further described with reference to specific embodiments, and the advantages and features of the invention will become apparent as the description proceeds. It should be understood that the examples are illustrative only and are not limiting upon the scope of the invention. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention, and that such changes and modifications may be made without departing from the spirit and scope of the invention.
The invention provides an immunoassay device for in vitro real-time determination of ABO and/or Rh blood type of a human, which comprises a plurality of sample holes, wherein a reaction membrane is respectively arranged below each sample hole, and each reaction membrane is respectively coated with an antibody for determining ABO blood type and Rh blood type, a positive quality control internal standard substance and a negative quality control internal standard substance for monitoring reaction effectiveness. The immunoassay device can simply, accurately and quickly determine the ABO and/or Rh blood type of a person through one-time operation, can effectively improve the detection efficiency and reduce the detection cost.
The immunoassay device of the present invention mainly comprises an upper cover, a reaction membrane, an absorbent pad, a backing and a base. For ease of storage and portability, the immunization device may further comprise a housing that completely encloses the cover and base. The provision of a mutually cooperating sealing structure between the upper cover housing and the base housing enables the upper cover housing and the base housing to be assembled together in a firmly fitting manner, the sealing structure preferably being designed as a male-female structure. The upper cover and the base can be respectively integrated with the respective shell, the edge of the upper cover is provided with a protrusion, and the edge of the base is provided with a groove corresponding to the protrusion of the upper cover, so that the upper cover and the base can be assembled together through the stable matching of the protrusion and the groove.
FIG. 1 shows an embodiment of the external structure of the immunoassay device for in vitro real-time determination of ABO and/or Rh blood type in a human body according to the present invention. The shape of the device may be any suitable shape, preferably a square stack, with a length of 80mm, a width of 40mm and a thickness of 3.5 to 5mm, and the material of the cover and base may be any of the widely used organic or inorganic materials, natural or artificial materials or any polymeric synthetic material, preferably a polypropylene material. Fig. 1 shows that 5 sample wells are provided in the immunoassay device, and for convenience of description, they are referred to as a first assay well 101, a positive quality control internal standard well 102, a second assay well 103, a negative quality control internal standard well 104, and a third assay well 105, respectively. It should be understood that the number of sample wells is not limited to 5, and other numbers of sample wells may be flexibly provided as necessary.
FIG. 2 shows a specific embodiment of the structure of the upper cover of the immunoassay device, in which the sample wells are provided in the upper cover, and a blood sample to be tested and a buffer are dropped through the sample wells, and the blood sample to be tested reacts with an antibody or a quality control internal standard substance coated on a reaction membrane below the corresponding sample well, and the reaction result can be directly observed visually through the sample wells.
In order to facilitate the distinction and observation of the results, in the upper cover shown in fig. 2, the sample wells are arranged in two rows, specifically: the first measuring hole 101, the second measuring hole 103 and the third measuring hole 105 for measuring blood type are arranged in a row, and the positive quality control internal standard hole 102 and the negative quality control internal standard hole 104 are arranged in a row. It will be appreciated that the sample wells on the immunoassay device of the present invention are not limited to this arrangement, and may be arranged in other suitable forms as desired.
The shape of the sample aperture can be designed and adjusted as desired. The first, second and third measurement holes 101, 103 and 105 preferably have circular upper and lower ports, respectively, the upper port having a diameter equal to or greater than the diameter of the lower port, wherein the upper port preferably has a diameter of 10.42mm and the lower port preferably has a diameter of 4.8 mm. The edge of the upper port is connected to the edge of the lower port by a gentle continuous face. The internal control standard hole 102 and the internal control standard hole 104 preferably have a circular upper port and a square lower port respectively, and the projection of the respective upper ports towards the lower port can completely cover the lower port, the diameter of the upper port is preferably 10.42mm, and the side length of the lower port is preferably 4 mm. The edge of the upper port is connected to the edge of the lower port by a gentle continuous face. The upper port, the lower port and the gentle and continuous surface of each sample hole form a liquid containing space.
FIG. 3 shows one embodiment of the structure of the reaction membrane, absorbent pad and backing of the immunoassay device. The reaction membranes 202 are arranged below the sample holes, and the antibodies for determining blood type or the positive quality control internal standard substance and the negative quality control internal standard substance for monitoring the reaction effectiveness are respectively coated on the reaction membranes 202. The material for preparing the reaction membrane 202 is selected from materials that allow agglutination reaction of red blood cells in the blood sample with antibodies on the membrane, and is preferably a cellulose material. The reaction membrane 202 can be selected from any one of cotton paper, printing paper, blotting paper, nitrocellulose membrane, cellulose acetate membrane or glass fiber, and preferably glass fiber. Additionally, the reaction membrane 202 is free of plastic film, allowing the use of the belt or air side of the membrane. The pore size of the reaction membrane 202 is preferably 12 μm for better surface-to-volume ratio. The reaction membrane 202 may have various shapes to fit the size of the sample well, and the shape of the reaction membrane 202 is preferably rectangular, and the size thereof is preferably 6mm × 5 mm. The thickness of the reaction membrane 202 is preferably 1mm to 1.25 mm.
For convenience of description below, the reaction membrane 202 located below the first measurement well 101, the second measurement well 103 and the third measurement well 105 is referred to as a detection zone reaction membrane, and the reaction membrane located below the positive quality control internal control standard well 102 and the negative quality control internal control standard well 104 is referred to as a quality control zone reaction membrane.
When a blood sample to be detected is dripped on the surface of the reaction membrane through each sample hole, the surface antigen of Red Blood Cells (RBC) in the blood sample and the corresponding antibody coated on the reaction membrane generate immunoreaction. When the antibodies on the detection area reaction membrane are anti-A antibodies and anti-B antibodies, the ABO blood type of the blood sample can be detected. Wherein, the anti-A antibody and the anti-B antibody are preferably any one of IgM antibody, IgG antibody or a mixture of IgM and IgG antibody, respectively. When the antibody in the detection area reaction membrane is an antibody for identifying Rh blood type, the Rh blood type of the blood sample can be detected. Among them, the antibody for identifying Rh blood group is preferably any one of an anti-D antibody, an anti-C antibody and an anti-e antibody. In addition, the anti-a antibody is preferably an anti-a monoclonal antibody, the anti-B antibody is preferably an anti-B monoclonal antibody, and the antibody for identifying Rh blood group is preferably an anti-D monoclonal antibody.
The quality control area reaction membrane below the positive quality control internal standard hole 102 is coated with a positive quality control internal standard substance, and the positive quality control internal standard substance is a substance capable of binding and reacting with human erythrocytes, and preferably an anti-human RBC antibody. A negative quality control internal standard substance is coated on the quality control area reaction membrane below the negative quality control internal standard hole 104, and the negative quality control internal standard substance is a substance which does not react with human erythrocytes in a binding way, and is preferably an anti-goat IgG antibody.
Corresponding antibodies, positive quality control internal standard substances and/or negative quality control internal standard substances can be coated on each reaction membrane by adopting a physical adsorption method, a competitive combination method or a magnetic bead immobilization method.
According to a preferred embodiment, the antibodies coated on the detection area reaction membrane for determining ABO blood group include anti-a monoclonal antibody and anti-B monoclonal antibody, the antibody for determining Rh blood group is anti-D monoclonal antibody, the positive quality control internal standard substance coated on the quality control area reaction membrane is anti-human RBC antibody, and the negative quality control internal standard substance is anti-goat IgG antibody. When coating the above antibodies, the antibodies are first diluted with a diluent, preferably TE buffer solution with pH 7.5, anti-A monoclonal antibody, anti-B monoclonal antibody and anti-D monoclonal antibody at a ratio of 1:1, and anti-RBC antibody and anti-goat IgG antibody at a ratio of 1: 5. Each antibody after dilution is then spotted on the reaction membrane individually, the volume of the spot is preferably 1. mu.L/mm, the shape of the spot is preferably circular, and the diameter thereof is preferably 4.5mm to 5.5 mm. The spotted reaction membrane was then dried in a DH chamber at 25 ℃ for 4 hours without sealing the membrane until it was attached to the backing with an adhesive material.
The absorbent pads 201 are adjacent to the corresponding reaction membranes 202, and the absorbent pads 201 are arranged side by side with the reaction membranes 202 so as to absorb the excess waste liquid during the detection process. Each reaction membrane 202 may be located on the same continuous membrane or may be a plurality of separate membranes. When the reaction membrane 202 includes a plurality of independent membranes, the absorbent pad 201 and the reaction membrane 202 are preferably arranged at intervals such that the reaction membranes 202 under adjacent sample wells are spaced apart from each other.
The absorbent pad 201 is a heavy media, preferably a cellulosic or cotton linters material, and may be selected from any of filter paper, cellulose fiber, and tissue paper. The absorbent pad 201 preferably has 99.2mg/cm2Can limit the filtration or flow of absorbed liquid through the reaction membrane 202. The thickness of the absorbent pad 201 is preferably 1.5 to 2.0mm, and the capillary flow rate is preferably 63.3s/4 cm. The absorbent pad 201 may be cut into different shapes defining dimensions to fit with an immunoassay device. The shape of the absorbent pad 201 is preferably rectangular, and the size of the absorbent pad 201 between two adjacent reaction membranes 202 is preferably 15mm × 5mm, and the size of 2 absorbent pads 201 positioned outside the reaction membranes 202 at both ends is preferably 7mm × 5mm and 8 × 5mm, respectively.
The backing is disposed on the opposite side of the upper cover with respect to the reaction membrane 202 and the absorbent pad 201, and the upper surface of the backing and the lower surfaces of the reaction membrane 202 and the absorbent pad 201 are adhered to each other, thereby fixing the positions of the reaction membrane 202 and the absorbent pad 201 and ensuring that the reaction membrane 202 is positioned below the corresponding sample well. The shape of the backing is adapted to the shape of the reactive film 202 and the absorbent pad 201, and is preferably rectangular. The detection zone backing 204 is preferably 30mm x 8mm in size and the quality control zone backing 205 is preferably 30mm x 5mm in size.
The backing is made of plastic materials, and polyethylene materials or PVC with the thickness of 1-1.5 mm are preferred. The number of backings can be selected based on the number and arrangement of sample wells. When the measurement apertures are arranged in a row with the internal positive and negative control apertures arranged in another row, the backings are preferably at least two, including at least one detection zone backing 204 for corresponding to the measurement apertures arranged in a row and at least one control zone backing 205 for corresponding to the internal positive and negative control apertures arranged in a row. On the detection zone backing 204, the absorbent pad 201 and the reaction membrane 202 are preferably arranged side by side in three-phase spaced arrangement, and from one end of the detection zone backing 204, the absorbent pad 201, the reaction membrane 202, and the absorbent pad 201 are arranged in this order. On the quality control zone backing 205, the absorption pad 201 and the reaction film 202 are preferably arranged side by side in two spaced relation, and the absorption pad 201, the reaction film 202, the absorption pad 201, the reaction film 202 and the absorption pad 201 are arranged in this order from one end of the quality control zone backing 205.
FIG. 4 shows one embodiment of a base structure of an immunoassay device. The base is in contact with the lower surface of the backing and is capable of supporting and holding the backing, thereby further positioning the reaction membrane 202 and the sample wells.
The upper surface of the base is provided with a backing support rib 303, a backing pressure buffering rib 302 and a backing fixing rib, wherein the three ribs are the backing fixing rib, the backing support rib 303 and the backing pressure buffering rib 302 in sequence from high to low in height. The number of the backing support ribs 303 may be set according to the length, width and number of the backing, wherein the top of at least one backing support rib 303 contacts the lower surface of the backing to support the backing. In order to ensure the stability of the backing, the top of the backing support rib 303 is preferably designed to be square, and the height of the top of the backing support rib 303 is preferably 1mm lower than the sample hole. The backing pressure cushioning ribs 302 and the backing support ribs 303 are arranged in parallel in the length direction of the backing, and preferably, the two ribs are staggered with each other. When the backing is deformed by the pressure of the reaction film, the absorption liner and the blood sample to be detected above, the backing pressure buffer rib 302 is in contact with the lower surface of the backing so as to better keep the upper surface of the backing in a horizontal state, prevent the liquid sample and the buffer from leaking among sample holes, further ensure that the reaction in each sample hole is not influenced mutually, and the height of the backing pressure buffer rib 302 is preferably 1 mm.
And backing fixing ribs are arranged around the side faces of the backing to clamp and fix the backing, and the backing fixing ribs comprise a first backing fixing rib 301 and a second backing fixing rib 304. The second backing fixing ribs 304 are located on the left and right sides of the backing support rib 303 and are arranged in pairs for fixing the left and right sides of the backing. The backing fixing ribs one 301 are positioned at the upper and lower sides of the backing support rib 303 and arranged in pairs for fixing the upper and lower sides of the backing.
In assembling the immunoassay device of the present invention, the detection zone backing 204 is first attached to the backing support ribs 303 of the base, and the detection zone backing 204 is secured by backing securing ribs 301 and 304. The control region backing 205 is then mounted on the support ribs 303 of the base plate and the control region backing 205 is fixed with backing fixing ribs 301 and 304. Next, the reaction membrane and the absorbent pad were arranged. Then, the upper cover is mounted, the position and state of the corresponding reaction membrane 202 are visually checked and/or adjusted from each sample well, and the upper cover and the base are mounted together by pressing the upper cover toward the base so that the protrusions of the edge of the upper cover are fitted into the corresponding grooves of the base, thereby forming the immunoassay device of the present invention.
According to a preferred embodiment of the present invention, the antibodies coated on the reaction membrane 202 for determining ABO blood group include an anti-a monoclonal antibody and an anti-B monoclonal antibody, the antibody for determining Rh blood group is an anti-D monoclonal antibody, the positive quality control internal standard substance is an anti-human RBC antibody, and the negative quality control internal standard substance is an anti-goat IgG antibody. When the immunoassay device is used to determine the ABO and/or Rh blood type of a human, it is preferable to use a mixed solution of 2% SDS solution and PBS solution as a washing buffer, the pH of the mixed solution is preferably 7.5 to 8.0, and 50. mu.L of the mixed solution is preferably dropped into each sample well. The measuring device is horizontally placed on the surface of a flat and clean support, 10uL of blood sample to be measured is respectively dripped into each sample hole, after the reaction membrane is fully soaked, 50uL of washing buffer solution is dripped, after the reaction membrane is fully soaked by the washing buffer solution, the result is observed, and the reaction membrane can be fully soaked within 2 minutes usually.
Fig. 5 shows a reaction principle of blood group determination using the immunoassay device of fig. 1, after a blood sample 401 is dropped into a sample well, the blood sample infiltrates and permeates into the reaction membrane 202, and the antibodies 203 on the reaction membrane 202 immunoreact with the corresponding antigens on the surfaces of red blood cells in the blood sample 401, thereby capturing and agglutinating the red blood cells, so that the reaction membrane 202 shows red color.
To help better understand the technical solution of the present invention, an example is provided below to illustrate the specific structure and application effect of the immunoassay device for in vitro real-time determination of ABO and/or Rh blood group in humans according to the present invention, wherein the immunoassay device used in the example is provided with five sample wells.
Examples
The immunoassay device in the embodiment is in a cuboid shape of 80mm multiplied by 40mm multiplied by 5mm, and five sample holes are arranged on the upper cover of the immunoassay device, wherein the five sample holes comprise three measuring holes, one positive quality control internal standard hole and one negative quality control internal standard hole. Wherein three survey holes are arranged into one row, and the last port and the lower port of every survey hole are circular, and the diameter of going up the port is 10.42mm, and the diameter of lower port is 4.8mm, passes through gentle face connection between the edge of going up the port and the edge of lower port. The positive quality control internal standard holes and the negative quality control internal standard holes are arranged in a row, the upper ports of the positive quality control internal standard holes and the negative quality control internal standard holes are circular with the diameter of 10.42mm, the lower ports of the positive quality control internal standard holes and the negative quality control internal standard holes are square with the side length of 4mm, and the edge of the upper ports is connected with the edge of the lower ports through a smooth surface.
Reaction films are respectively arranged under the three measuring holes, and the reaction films are respectively coated with an anti-A monoclonal antibody, an anti-B monoclonal antibody and an anti-D monoclonal antibody, and for identification, A, B and D are respectively marked beside the corresponding measuring holes on the upper cover. An anti-human RBC antibody is coated on the reaction membrane below the positive quality control internal standard hole to serve as a positive quality control internal standard substance, and a corresponding position on the upper cover is marked with PC. The reaction membrane below the negative quality control internal standard hole is coated with an anti-goat IgG antibody as a negative quality control internal standard substance, and the corresponding position on the upper cover is marked with NC.
Absorbent pads are respectively arranged on both sides of the reaction film corresponding to the three measurement holes, and the reaction film comprises four absorbent pads and three reaction films in total, which are attached side by side on the upper surface of the detection area backing with the size of 30mm × 8mm in the order of the absorbent pads → the reaction film → the absorbent pads. And the two sides of each reaction film below the positive quality control internal standard hole and the negative quality control internal standard hole are respectively provided with an absorption liner, the absorption liners and the two reaction films are totally included, and the absorption liners and the reaction films are attached to the upper surface of a 30mm multiplied by 5mm quality control area back lining side by side according to the sequence of the absorption liner → the reaction film → the absorption liner.
The detection zone back lining is fixed on the base through four back lining pressure buffering ribs, three back lining supporting ribs, a pair of upper and lower back lining fixing ribs and three pairs of left and right back lining fixing ribs. The back lining of the quality control area is fixed on the base through three back lining pressure buffering ribs, two back lining supporting ribs, a pair of upper and lower back lining fixing ribs and two pairs of left and right back lining fixing ribs.
FIG. 6 is a graph showing the result judgment in the blood type test using the immunoassay device of the present embodiment.
As shown by reference numeral 601 in FIG. 6, when the measurement hole A, the measurement hole D and the positive quality control internal standard hole PC show coloring spots (red will be observed by naked eyes in practice), and the measurement hole B and the negative quality control internal standard hole NC do not show coloring, the blood type of the blood sample to be detected is judged to be Rh type A+。
As shown by reference numeral 602 in fig. 6, when the measurement hole a and the positive quality control internal standard hole PC show colored spots (red will be observed by naked eyes in practice), and the measurement hole B, the measurement hole D and the negative quality control internal standard hole NC do not show coloring, the blood type of the blood sample to be tested is determined to be Rh type a-。
As shown by reference numeral 603 in fig. 6, when the measurement hole B, the measurement hole D and the positive quality control internal standard hole PC show colored spots (red will be observed by naked eyes in practice), and the measurement hole a and the negative quality control internal standard hole NC do not show coloring, the blood type of the blood sample to be tested is judged to be Rh type B+。
As shown by reference numeral 604 in fig. 6, when the measurement hole B and the positive quality control internal standard hole PC show colored spots (red will be observed by naked eyes in practice), and the measurement hole a, the measurement hole D and the negative quality control internal standard hole NC do not show coloring, the blood type of the blood sample to be tested is judged to be Rh type B-。
As shown by reference numeral 605 in FIG. 6, colored spots (macroscopically) appear in the measurement well A, the measurement well B, the measurement well D and the positive quality control internal control well PCRed actually observed), and when the NC hole does not appear coloring, the blood type of the blood sample to be detected is judged to be AB type Rh+。
As shown by reference numeral 606 in fig. 6, when the measurement hole a, the measurement hole B and the positive quality control internal standard hole PC show colored spots (red will be observed by naked eyes in practice), and the measurement hole D and the negative quality control internal standard hole NC do not show coloring, the blood type of the blood sample to be tested is judged to be AB type Rh-。
As shown by reference numeral 607 in fig. 6, when the measurement hole D and the positive quality control internal standard hole PC show colored spots (red will be observed by naked eyes in practice), and the measurement hole a, the measurement hole B and the negative quality control internal standard hole NC do not show coloring, it is determined that the blood type of the blood sample to be measured is O-type Rh+。
As shown by reference numeral 608 in fig. 6, when the positive quality control internal standard hole PC shows a colored spot (red will be observed by naked eyes), and the measurement hole a, the measurement hole B, the measurement hole D and the negative quality control internal standard hole NC do not show coloring, it is determined that the blood type of the blood sample to be tested is O-type Rh-。
As shown by reference numeral 609 in fig. 6, when the measurement well a, the measurement well B, the measurement well D, and the positive quality control internal standard hole PC do not show colored spots, but the negative quality control internal standard hole NC shows colored spots, it is determined that the detection is invalid, and a new immunoassay device needs to be replaced for detection again.
The above description is only a preferred embodiment of the present invention, and the protection scope of the present invention is not limited to the above embodiments, and all technical solutions belonging to the idea of the present invention belong to the protection scope of the present invention. It will be apparent to those skilled in the art that improvements and modifications may be made without departing from the principles of the invention and are intended to be within the scope of the invention.
Claims (10)
1. An immunoassay device for in vitro real-time determination of the ABO and/or Rh blood group of a human, said immunoassay device comprising a cover, a reaction membrane, an absorbent pad, a backing and a base, characterized in that:
the upper cover is provided with a plurality of sample holes, a reaction film is respectively arranged below each sample hole, the reaction film below each sample hole is respectively coated with an antibody for determining blood type or a positive quality control internal standard substance and a negative quality control internal standard substance for monitoring reaction effectiveness, the absorption pad and the reaction films are arranged side by side at intervals so that the reaction films below adjacent sample holes are spaced,
the back lining is positioned on the opposite side of the upper cover relative to the reaction film and the absorption liner, one surface of the back lining is attached to the reaction film and the absorption liner, the other surface of the back lining is in contact with the base and is supported and fixed by the base, and the base and the upper cover are assembled together in a stable fit manner.
2. The immunoassay device of claim 1, wherein:
the plurality of sample holes comprise measuring holes, positive quality control internal standard holes and negative quality control internal standard holes, the positive quality control internal standard substance is an anti-human RBC antibody, and the negative quality control internal standard substance is an anti-goat IgG antibody.
3. The immunoassay device of claim 2, wherein:
the kit comprises three measuring holes, and anti-A antibodies, anti-B antibodies and antibodies for identifying Rh blood types are coated on corresponding reaction membranes below the three measuring holes respectively.
4. The immunoassay device of claim 3, wherein:
the three measurement holes are arranged in a row, and the positive quality control internal standard hole and the negative quality control internal standard hole are arranged in a row.
5. The immunoassay device of any one of claims 2 to 4, wherein;
the measuring hole is provided with an upper port and a lower port which are circular, the diameter of the upper port is larger than that of the lower port, and the edge of the upper port is connected with the edge of the lower port through a smooth continuous surface.
6. The immunoassay device of any one of claims 2 to 4, wherein:
the positive quality control internal standard hole and the negative quality control internal standard hole are both provided with a circular upper port and a square lower port, the projection of the upper port vertically facing the lower port can completely cover the lower port, and the edge of the upper port is connected with the edge of the lower port through a smooth and continuous surface.
7. The immunoassay device of claim 3, wherein:
the anti-A antibody and the anti-B antibody are respectively selected from any one of IgM antibodies, IgG antibodies or a mixture of IgM and IgG antibodies, and the antibody for identifying Rh blood group is selected from any one of anti-D antibodies, anti-C antibodies, anti-C antibodies and anti-e antibodies.
8. The immunoassay device of claim 1, wherein:
the substrate of the reaction membrane is selected from any one of cotton paper, printing paper, blotting paper, nitrocellulose membrane and cellulose acetate membrane or any combination thereof.
9. The immunoassay device of claim 1, wherein:
the absorption pad is selected from one or any combination of filter paper, cellulose fiber and paper towel.
10. The immunoassay device of claim 1, wherein:
the edge of the upper cover is provided with a protrusion, the edge of the base is provided with a groove, and the upper cover and the base can be assembled together through the stable matching of the protrusion and the groove.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011015285.8A CN112345773A (en) | 2020-09-24 | 2020-09-24 | Immunoassay device for in vitro instant determination of ABO and/or Rh blood type of human |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011015285.8A CN112345773A (en) | 2020-09-24 | 2020-09-24 | Immunoassay device for in vitro instant determination of ABO and/or Rh blood type of human |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112345773A true CN112345773A (en) | 2021-02-09 |
Family
ID=74357690
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011015285.8A Withdrawn CN112345773A (en) | 2020-09-24 | 2020-09-24 | Immunoassay device for in vitro instant determination of ABO and/or Rh blood type of human |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112345773A (en) |
Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1337580A (en) * | 2000-08-08 | 2002-02-27 | 清华大学 | Solid molecule operating method in microfluid system |
| CN1766616A (en) * | 2005-10-17 | 2006-05-03 | 中国人民解放军第三军医大学第一附属医院 | With colloid gold label clq the give instruction protein chip/microarray detection system and the detection method of agent |
| CN1975423A (en) * | 2006-09-06 | 2007-06-06 | 浙江清华长三角研究院 | Immuno magnetic bead and producing method, and method and test plate for detection |
| CN101169414A (en) * | 2007-08-09 | 2008-04-30 | 嘉兴博泰生物科技发展有限公司 | Water body Chlamydomonas reinhaidtii toxin detection method |
| CN203287379U (en) * | 2013-05-13 | 2013-11-13 | 艾博生物医药(杭州)有限公司 | Detection board for placing test strip |
| CN104769429A (en) * | 2012-06-11 | 2015-07-08 | Abo血型诊断公司 | In vitro diagnosis device and uses thereof |
| CN205333656U (en) * | 2016-01-21 | 2016-06-22 | 浙江东方基因生物制品有限公司 | Economic environmental protection test pregnant stick |
| CN109459575A (en) * | 2018-09-19 | 2019-03-12 | 厦门为正生物科技股份有限公司 | A kind of ABO&RhD blood group detection device |
| CN208847748U (en) * | 2018-09-25 | 2019-05-10 | 深圳市正海生物科技有限公司 | A kind of immunochromatographydetection detection card |
| CN210269872U (en) * | 2019-06-27 | 2020-04-07 | 英科新创(厦门)科技股份有限公司 | Human blood group antigen detection device |
| CN211348250U (en) * | 2019-10-31 | 2020-08-25 | 杭州奥泰生物技术股份有限公司 | Biodegradable detection device |
| CN111638373A (en) * | 2020-06-08 | 2020-09-08 | 长春博德生物技术有限责任公司 | Kit for ABO blood group positive typing and Rh blood group detection and preparation method and detection method thereof |
-
2020
- 2020-09-24 CN CN202011015285.8A patent/CN112345773A/en not_active Withdrawn
Patent Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1337580A (en) * | 2000-08-08 | 2002-02-27 | 清华大学 | Solid molecule operating method in microfluid system |
| CN1766616A (en) * | 2005-10-17 | 2006-05-03 | 中国人民解放军第三军医大学第一附属医院 | With colloid gold label clq the give instruction protein chip/microarray detection system and the detection method of agent |
| CN1975423A (en) * | 2006-09-06 | 2007-06-06 | 浙江清华长三角研究院 | Immuno magnetic bead and producing method, and method and test plate for detection |
| CN101169414A (en) * | 2007-08-09 | 2008-04-30 | 嘉兴博泰生物科技发展有限公司 | Water body Chlamydomonas reinhaidtii toxin detection method |
| CN104769429A (en) * | 2012-06-11 | 2015-07-08 | Abo血型诊断公司 | In vitro diagnosis device and uses thereof |
| CN203287379U (en) * | 2013-05-13 | 2013-11-13 | 艾博生物医药(杭州)有限公司 | Detection board for placing test strip |
| CN205333656U (en) * | 2016-01-21 | 2016-06-22 | 浙江东方基因生物制品有限公司 | Economic environmental protection test pregnant stick |
| CN109459575A (en) * | 2018-09-19 | 2019-03-12 | 厦门为正生物科技股份有限公司 | A kind of ABO&RhD blood group detection device |
| CN208847748U (en) * | 2018-09-25 | 2019-05-10 | 深圳市正海生物科技有限公司 | A kind of immunochromatographydetection detection card |
| CN210269872U (en) * | 2019-06-27 | 2020-04-07 | 英科新创(厦门)科技股份有限公司 | Human blood group antigen detection device |
| CN211348250U (en) * | 2019-10-31 | 2020-08-25 | 杭州奥泰生物技术股份有限公司 | Biodegradable detection device |
| CN111638373A (en) * | 2020-06-08 | 2020-09-08 | 长春博德生物技术有限责任公司 | Kit for ABO blood group positive typing and Rh blood group detection and preparation method and detection method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US4623461A (en) | Transverse flow diagnostic device | |
| US6514769B2 (en) | Multiple analyte assay device with sample integrity monitoring system | |
| CN1125983C (en) | Analytical device for membrane-based assays | |
| US5084245A (en) | Assay device for swab borne analytes | |
| US7347972B1 (en) | Multiple analyte assay device | |
| US5147780A (en) | Multiwell stat test | |
| EP0319294A2 (en) | Improved membrane assay using focused sample application | |
| WO2003079018A1 (en) | Drug test kit | |
| JP2676108B2 (en) | Immunoenzymatic detection of substances from blood drops or fluids from any biological medium | |
| CN106645675B (en) | The detection reagent of urea nitrogen and the Test paper of urea nitrogen | |
| EP0767909B1 (en) | Multi-layer test devices and methods of assaying for fructosamine | |
| US20040081581A1 (en) | Urine specimen cup toxicology indicator cap | |
| EP0239174B1 (en) | Biological diagnostic device | |
| CN1849514A (en) | Particle agglutination detection method and device | |
| US8623291B2 (en) | Multiple analyte assay device | |
| KR20220025857A (en) | Blood group antigen detection component | |
| CN112345773A (en) | Immunoassay device for in vitro instant determination of ABO and/or Rh blood type of human | |
| CN110249221A (en) | For detecting the vertical current dynamic formula test device of the concentration of glucose in fluid sample | |
| JP3007410B2 (en) | Multiwell test | |
| WO2020161238A1 (en) | Lateral flow device | |
| US20200341005A1 (en) | Carrier and method for detecting an analyte in dried blood spots | |
| US20210349083A1 (en) | CORONAVIRUS IgG/IgM MULTIPLEXED DUAL PATH IMMUNOASSAY DEVICE | |
| AU2003262462B2 (en) | Multiple analyte assay device |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WW01 | Invention patent application withdrawn after publication | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20210209 |