Automatic test card for multi-blood group system and test method

Patent metadata

Field	Value
Publication number	US-12571804-B2
Application number	US-202117904601-A
Country	US
Kind code	B2
Filing date	Mar 3, 2021
Priority date	Mar 4, 2020
Publication date	Mar 10, 2026
Grant date	Mar 10, 2026

Abstract

The present disclosure provides an automatic test card for a multi-blood group system and a test method. The test card of a multi-blood group system includes four parts from top to bottom: a sample loading layer, a reaction layer, a color developing layer, and an identification layer. The automatic test card for a multi-blood group system and the test method combines blood group detection with encoding, and can rapidly and automatically identify and test multiple blood groups, reduce manual judgment errors, and improve detection efficiency. The method does not require special equipment and achieves rapid, accurate, automated, and portable detection of the blood groups.

First claim

What is claimed is:

1 . A test card of a multi-blood group system, comprising four layers from top to bottom: a sample loading layer, a reaction layer, a color developing layer, and an identification layer;
the sample loading layer being provided with a sample loading hole, the reaction layer being provided with a whole blood pad or a filter pad for filtering red blood cells (RBCs) from whole blood, and bottom of the sample loading hole being connected to the whole blood pad or the filter pad; an end of the whole blood pad being connected to a plurality of antibody pads pre-coated with blood group antibodies, or an end of the filter pad being connected to antigen pads pre-coated with blood group antigens via a delay pad; the reaction layer being independently provided with a quality control strip; wherein the color developing layer is provided with a plurality of color developing areas connected to an end of the antibody pad or the antigen pad; the identification layer is reversely connected to the color developing layer, and comprises three two-dimensional code positioning areas, a calibration area, a plurality of test areas, and a subject information area; and positions of the test areas and positions of the color developing areas being aligned with each other one-to-one such that the test areas overlaps with the color developing areas and provides a hollow hole or a transparent membrane on the test areas; and the identification layer has a shape of square, the three two-dimensional code positioning areas are distributed in three corners of the square, and the subject information area includes a QR two-dimensional code, a three-dimensional code, or a barcode used for registration and storage of subject information and is distributed in the fourth corner of the square; and the calibration area is composed of 6 color blocks, with RGB being (68, 114, 196), (240, 142, 24), (44, 220, 65), (218, 46, 148), (220, 78, 44), and (41, 219, 223).

2 . The test card according to claim 1 , wherein the test card has a three-dimensional or two-dimensional structure.

3 . The test card according to claim 1 , wherein the sample loading layer is made of glass fiber, cotton pulp paper, napkin, filter paper, gauze, or hydrogel, the sample loading hole is round or square, and the sample loading layer is treated using a technology selected from wax printing and photolithography to form a hydrophilic area and a hydrophobic area; the whole blood pad, the antibody pad, the delay pad, and the antigen pad each are made of a material selected from non-woven fabrics, non-woven paper, filter paper, cotton pulp paper, and glass fiber with a pore size of 8 μm to 20 μm; and the filter pad is made of Prussian blue membrane, glass fiber, graphene cloth, carbon cloth, carbon paper, whole blood separation membrane, or nitrocellulose membrane.

4 . The test card according to claim 1 , wherein the antibody pad is pre-coated with commercialized ABO, Rh, MNS, Kell, P, Kidd, Duffy blood group antibodies which are immobilized by freeze-drying at 80° C., plasma treatment, covalent bond coupling, vacuum drying at 20° C. to 50° C., immunomagnetic bead binding, or electrostatic adsorption; and the antigen pad is pre-coated with ABO blood group antigens selected from A1, A2, B, and O RBCs; and the ABO blood group antigens are immobilized by freeze-drying at 80° C., vacuum drying at 4° C. to 30° C., immunomagnetic bead binding, or electrostatic adsorption; or alternatively, fresh RBCs are used as the antigen and added directly on the antigen pad.

5 . The test card according to claim 1 , wherein the delay pad achieves delay chemically by pre-coating with saccharides, paraffin, or alkyl ketene dimers, or physically by changing the shape of the delay pad or increasing the length and width of the delay pad.

6 . The test card according to claim 1 , wherein the color developing layer is made of nitrocellulose membrane, cellulose acetate membrane, or polyester cellulose membrane; color developing is performed on the color developing layer by a method selected from color change and gray value change; each of the color developing areas is any shape of a square, a rectangle, or a circle; the pre-coated antibody pads or the pre-coated antigen pads are pre-coated with nanoparticles or a dye, and the nanoparticles are selected from latex particles, gold nanoparticles, and silver nanoparticles, and the dye is selected from a biuret reagent, methyl green, pyronin, a bromothymol blue solution, and ninhydrin.

7 . The test card according to claim 1 , wherein the identification layer is made of printing paper, cotton paper, or blotting paper, and the positioning area, the calibration area, and the subject information area are visualized by wax printing, laser printing, or photosensitive seal; the positioning area is used for automatic positioning and deviation correction in automatic detection; the calibration area is used for color calibration to prevent interference from external environment and a photographing instrument.

8 . The test card according to claim 1 , wherein the hollow structure is formed by laser cutting, chemical etching, or manual cutting, and the transparent membrane is made of polyvinyl chloride, polyethylene, polypropylene, polystyrene, or resin.

9 . An automatic test method for a multi-blood group system using the test card according to claim 1 , comprising the following steps: adding whole blood to the sample loading hole, after 30 sec to 1 min adding PBS, normal saline, or ultrapure water to the sample loading hole, observing a color change in the test area with naked eyes after 10 sec, and interpreting results with automatic identification software; wherein the automatic identification software is capable of being installed in a common electronic device comprising a smartphone, a computer, and a scanner;
wherein the test card comprises four layers from top to bottom: a sample loading layer, a reaction layer, a color developing layer, and an identification layer; wherein the sample loading layer is provided with a sample loading hole, the reaction layer is provided with a whole blood pad or a filter pad for filtering red blood cells (RBCs) from whole blood, and bottom of the sample loading hole is connected to the whole blood pad or the filter pad; an end of the whole blood pad is connected to a plurality of antibody pads pre-coated with blood group antibodies, or an end of the filter pad is connected to antigen pads pre-coated with blood group antigens via a delay pad; the reaction layer is independently provided with a quality control strip; wherein the color developing layer is provided with a plurality of color developing areas connected to an end of the antibody pad or the antigen pad; the identification layer is reversely connected to the color developing layer, and comprises three two-dimensional code positioning areas, a calibration area, a plurality of test areas, and a subject information area; and positions of the test areas and positions of the color developing areas being aligned with each other one-to-one such that the test areas overlaps with the color developing areas and provides a hollow hole or a transparent membrane on the test areas; and the identification layer has a shape of square, the three two-dimensional code positioning areas are distributed in three of corners of the square, and the subject information area includes a QR two-dimensional code, a three-dimensional code, or a barcode used for registration and storage of subject information and is distributed in the fourth corner of the square; and the calibration area is composed of 6 color blocks, with RGB being (68, 114, 196), (240, 142, 24), (44, 220, 65), (218, 46, 148), (220, 78, 44), and (41, 219,

Assignees

• Univ Chongqing

Inventors

• Luo Yang
• Zhang Hong

Classifications

• G01N33/558

  using diffusion or migration of antigen or antibody {(immunochromatographic test strips G01N33/54387)} · CPC title

• G01N33/531

  Production of immunochemical test materials · CPC title

• G01N33/54391

  based on vertical flow · CPC title

• G01N33/80 Primary

  involving blood groups or blood types {or red blood cells (white blood cells G01N33/56972)} · CPC title