A dry test strip for reverse typing of ABO blood type based on artificial antigen

Abstract

The present invention discloses an ABO blood type reverse typing dry test strip based on artificial antigens, which is characterized by comprising a bottom plate and a sample adding pad, a separation pad, an antigen pad, a first interception pad, a second interception pad, a third interception pad and a water absorption pad which are sequentially connected and arranged on the bottom plate. The ABO blood type reverse typing dry test strip based on artificial antigens of the present invention is convenient to store, can be stored at room temperature (2-30°C), has a long validity period (not less than 12 months), is convenient to carry, is simple to operate, does not require auxiliary equipment, directly detects whole blood, does not require serum separation, and the results are easy to interpret and objective and reliable. It is suitable for rapid detection of ABO blood types at the blood collection site, family self-testing of blood types, initial blood type screening in hospital testing departments, and emergency situations in the field.

Description

Technical Field

[0001] The invention belongs to the technical field of bioengineering, and in particular relates to an ABO blood type reverse typing dry test strip based on artificial antigens. Background Art

[0002] The ABO blood type system divides blood types into O, A, B and AB blood types based on the difference of A and B agglutinogens on red blood cells. Type A blood contains A antigen on red blood cells and anti-B antibodies in serum; Type B blood contains B antigen on red blood cells and anti-A antibodies in serum; Type O blood contains no A and B antigens on red blood cells and anti-A and anti-B antibodies in serum; Type AB blood contains both A and B antigens on red blood cells and no anti-A and anti-B antibodies in serum. A antigen and anti-A antibody, B antigen and anti-B antibody will produce immune binding reaction, causing red blood cells to agglutinate. ABO blood type is determined by both red blood cell surface antigens and antibodies in serum. Using standard anti-A and anti-B antibodies to detect red blood cell antigens is called positive typing; using standard A and B antigens to detect antibodies in serum is called negative typing. Only by testing positive and negative typing at the same time can the ABO blood type be correctly determined.

[0003] Correct blood typing is an important task to ensure the safety of clinical blood transfusion and save lives. ABO blood typing is one of the routine blood typing serological tests before blood transfusion. In order to ensure correct blood typing, the Ministry of Health issued a document in 2000 (Weiyi Fa

[2000] No. 184) which clearly stipulates that every individual who donates blood and receives blood must undergo ABO blood typing positively and negatively. At present, the most commonly used ABO blood type reverse typing identification method in clinical laboratories is the hemagglutination test (including test tube method, plate method, microcolumn gel method). The standard A, B and O type red blood cells used in the hemagglutination test have a short shelf life, generally stored for 3 months at 2-8°C, and are prone to microbial contamination and hemolysis. If the blood unit prepares it by itself, there is no unified standard, and the preparation method of each unit is different. The operation is cumbersome, and the composition and physical and chemical properties of the red blood cell preservation fluid are different, resulting in extremely unstable quality of the reverse typing red blood cell reagent, and a short in vitro shelf life. The test requires separation of serum and red blood cells (the test object is serum or plasma), and the reaction requires centrifugal equipment or glass slides. The operation is cumbersome, the result is difficult to interpret, and it is easy to cause errors in blood type identification, and even serious transfusion accidents. In addition, there is a magnetic bead method. Before using the magnetic bead method, the reagent needs to be redissolved. The storage time of the redissolved reagent is short, and it must be used up within 7 days in general, and a special magnetization program-controlled oscillator is required. Summary of the invention

[0004] The purpose of the present invention is to overcome the defects of the prior art and provide an ABO blood type reverse typing dry test strip based on artificial antigens.

[0005] The technical solution of the present invention is as follows:

[0006] An ABO blood type reverse typing dry test strip based on artificial antigens comprises a bottom plate and a sample adding pad, a separation pad, an antigen pad, a first interception pad, a second interception pad, a third interception pad and a water absorption pad which are sequentially connected and arranged on the bottom plate, wherein:

[0007] The separation pad is used to separate whole blood, retain RBCs, and allow plasma and serum to pass through;

[0008] The antigen pad fixes artificial A antigen or artificial B antigen labeled with tracer particles by physical adsorption or chemical cross-linking;

[0009] The pore size of the first interception pad is larger than the unagglutinated artificial antigen A or artificial antigen B, and smaller than the agglutinated complex of the artificial antigen A or artificial antigen B, and plays a role of preliminary interception to intercept larger agglutinated particles;

[0010] The pore size of the second interception pad is larger than the unagglutinated artificial antigen A or artificial antigen B, and smaller than the pore size of the agglutinated complex of the artificial antigen A or artificial antigen B and the first interception pad, so as to further intercept smaller agglutinated particles.

[0011] The pore size of the third interception pad is smaller than that of the unagglutinated artificial A antigen or artificial B antigen, and it plays the role of ultimate interception, intercepting the unagglutinated artificial antigen. A waterproof transparent tape is affixed thereon to prevent the unreacted antigen from flowing into the absorbent pad, and there is a gap between the waterproof transparent tape and the second interception pad as a result interpretation area.

[0012] In a preferred embodiment of the present invention, one end of the sample loading pad corresponds to one end of the bottom plate; one end of the separation pad is placed on the other end of the sample loading pad; one end of the antigen pad is placed on the other end of the separation pad; one end of the first interception pad is placed on the other end of the antigen pad; one end of the second interception pad is placed on the other end of the first antigen pad, and the other end of the second interception pad is placed on one end of the third interception pad; one end of the water absorbent pad is placed on the other end of the third interception pad; and the other end of the water absorbent pad corresponds to the other end of the bottom plate.

[0013] Further preferably, the base plate is made of polyester.

[0014] Further preferably, the material of the sample adding pad includes glass fiber.

[0015] Further preferably, the separation pad is a blood filter membrane, a nitrocellulose membrane or a cellulose acetate membrane.

[0016] Further preferably, the antigen pad is made of porous polymer material.

[0017] Further preferably, the first and second interception pads are made of glass fiber.

[0018] Further preferably, the third interception pad is a nitrocellulose membrane or a cellulose acetate membrane.

[0019] Further preferably, the absorbent pad is pulp paper.

[0020] The invention has the following beneficial effects: the ABO blood type reverse typing dry test strip based on artificial antigen is easy to store, can be stored at room temperature (2-30°C), has a long validity period (not less than 12 months), is easy to carry, simple to operate, does not require auxiliary equipment, can directly detect whole blood, does not require serum separation, and is easy to interpret and objective and reliable. It is suitable for rapid detection of ABO blood type at the blood collection site, family self-test of blood type, initial blood type screening in hospital testing departments, and emergency situations in the field. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 The figure is a schematic diagram of the structure of the ABO blood type reverse typing dry test strip of the present invention. DETAILED DESCRIPTION

[0022] The technical solution of the present invention is further illustrated and described below through specific implementation modes in combination with the accompanying drawings.

[0023] like Figure 1 As shown, an ABO blood type reverse typing dry test strip based on artificial antigens comprises a bottom plate 1 and a sample loading pad 2, a separation pad 3, an antigen pad 4, a first interception pad 5, a second interception pad 6, a third interception pad 7 and a water absorbent pad 8 which are sequentially connected and arranged on the bottom plate 1. Specifically, one end of the sample loading pad 2 corresponds to one end of the bottom plate 1; one end of the separation pad 3 is placed on the other end of the sample loading pad 2; one end of the antigen pad is placed on the other end of the separation pad 3; one end of the first interception pad 5 is placed on the other end of the antigen pad; one end of the second interception pad 6 is placed on the other end of the first antigen pad, and the other end of the second interception pad 6 is placed on one end of the third interception pad 7; one end of the water absorbent pad 8 is placed on the other end of the third interception pad 7; the other end of the water absorbent pad 8 corresponds to the other end of the bottom plate 1, wherein:

[0024] The separation pad 3 is used to separate whole blood, intercept RBCs, and allow plasma and serum to pass;

[0025] The antigen pad 4 fixes the artificial A antigen or artificial B antigen marked by the tracer particles by physical adsorption or chemical cross-linking; the preparation method of the artificial A antigen or artificial B antigen is a chemical synthesis method and an artificial extraction method, wherein the artificial extraction method includes a hypotonic swelling and crushing extraction method, an ultrasonic crushing extraction method, a freezing and crushing extraction method, an extraction method, etc., preferably a hypotonic swelling and crushing extraction method, see CN103033632A for details; the tracer particles include colloidal gold, colloidal selenium, colored latex, magnetic beads and gelatin, etc., wherein colloidal gold is preferred, and the combination mode of the colloidal gold and the artificial A antigen or the artificial B antigen is as shown in the colloidal gold physical adsorption method shown in CN103033632A;

[0026] The pore size of the first interception pad 5 is larger than the unagglutinated artificial antigen A or artificial antigen B, and smaller than the agglutinated complex of the artificial antigen A or artificial antigen B, and plays a role of preliminary interception to intercept larger agglutinated particles;

[0027] The pore size of the second interception pad 6 is larger than the unagglutinated artificial antigen A or artificial antigen B, and smaller than the pore size of the agglutinated complex of the artificial antigen A or artificial antigen B and the first interception pad 5, so as to further intercept smaller agglutinated particles.

[0028] The pore size of the third interception pad 7 is smaller than that of the unagglutinated artificial A antigen or artificial B antigen, and it plays the role of ultimate interception, intercepting the unagglutinated artificial antigen. A waterproof transparent tape 70 is affixed thereon to prevent the unreacted antigen from being lost to the absorbent pad 8, and there is a gap between the waterproof transparent tape 70 and the second interception pad 6 as a result reading area 71.

[0029] Preferred solution: the base plate 1 is made of polyester, the sample loading pad 2 is made of glass fiber, the separation pad 3 is a blood filter membrane, a nitrocellulose membrane or a cellulose acetate membrane, the antigen pad is made of a porous polymer material, the first and second interception pads 6 are made of glass fiber, the third interception pad 7 is a nitrocellulose membrane or a cellulose acetate membrane, and the absorbent pad 8 is pulp paper.

[0030] The method of use of the present invention is as follows:

[0031] The whole blood sample is dripped on one end of the sample pad 2 (close to the separation pad 3). The serum or plasma to be separated flows through the antigen pad and waits for a period of time, usually 2-3 minutes (the reaction time between the antibody in the serum and the antigen in the antigen pad). Then, the rinse solution is dripped on the other end of the sample pad 2, and the sample is waited for a period of time, usually 2-5 minutes, and the result is judged.

[0032] The interpretation method is: if the result interpretation area 71 is colored (artificial antigen color marked by tracer particles), it is a negative result, and if it is not colored, it is a positive result. For example: the antigen pad is coated with artificial A antigen (artificial A antigen test strip), and the result interpretation area 71 is colored, indicating that there is no anti-A antibody in the serum, and the result interpretation area 71 is not colored, indicating that there is anti-A antibody in the serum. The antigen pad is coated with artificial B antigen (B antigen test strip), and the result interpretation area 71 is colored, indicating that there is no anti-B antibody in the serum, and the result interpretation area 71 is not colored, indicating that there is anti-B antibody in the serum. Each sample needs to be tested with both A and B antigen test strips, and the result interpretation is shown in the following table:

[0033] Blood Type A B blood type O blood type AB blood type A strip (antigen pad coated with artificial A antigen) Color No color No color Color B strip (antigen pad coated with solidified artificial B antigen) No color Color No color Color

[0034] The above description is only a preferred embodiment of the present invention, and therefore cannot be used to limit the scope of the present invention. That is, equivalent changes and modifications made according to the patent scope of the present invention and the contents of the specification should still fall within the scope of the present invention.

Claims

1. An ABO blood type reverse typing dry test strip based on artificial antigen, characterized in that: It comprises a bottom plate and a sample adding pad, a separation pad, an antigen pad, a first interception pad, a second interception pad, a third interception pad and a water absorption pad which are sequentially connected and arranged on the bottom plate, wherein: The separation pad is used to separate whole blood, retain RBCs, and allow plasma and serum to pass; The antigen pad fixes artificial A antigen or artificial B antigen labeled with tracer particles by physical adsorption or chemical cross-linking; The pore size of the first interception pad is larger than the unagglutinated artificial antigen A or artificial antigen B, and smaller than the agglutinated complex of the artificial antigen A or artificial antigen B; The pore size of the second interception pad is larger than the pore size of the unagglutinated artificial antigen A or artificial antigen B, and smaller than the pore size of the agglutinated complex of the artificial antigen A or artificial antigen B and the first interception pad; The pore size of the third interception pad is smaller than that of the unagglutinated artificial A antigen or artificial B antigen, and a waterproof transparent tape is affixed thereon to prevent the unreacted antigen from flowing into the absorbent pad. There is a gap between the waterproof transparent tape and the second interception pad as a result reading area.

2. The ABO blood type reverse typing dry test strip according to claim 1, characterized in that: One end of the sample loading pad corresponds to one end of the bottom plate; one end of the separation pad is placed on the other end of the sample loading pad; one end of the antigen pad is placed on the other end of the separation pad; one end of the first interception pad is placed on the other end of the antigen pad; one end of the second interception pad is placed on the other end of the first antigen pad, and the other end of the second interception pad is placed on one end of the third interception pad; one end of the water absorbent pad is placed on the other end of the third interception pad; and the other end of the water absorbent pad corresponds to the other end of the bottom plate.

3. The ABO blood type reverse typing dry test strip according to claim 1 or 2, characterized in that: The material of the bottom plate is polyester.

4. The ABO blood type reverse typing dry test strip according to claim 1 or 2, characterized in that: The material of the sample adding pad includes glass fiber.

5. The ABO blood type reverse typing dry test strip according to claim 1 or 2, characterized in that: The separation pad is a blood filter membrane, a nitrocellulose membrane or a cellulose acetate membrane.

6. The ABO blood type reverse typing dry test strip according to claim 1 or 2, characterized in that: The material of the antigen pad is porous polymer material.

7. The ABO blood type reverse typing dry test strip according to claim 1 or 2, characterized in that: The first and second interception pads are made of glass fiber.

8. The ABO blood type reverse typing dry test strip according to claim 1 or 2, characterized in that: The third interception pad is a nitrocellulose membrane or a cellulose acetate membrane.

9. The ABO blood type reverse typing dry test strip according to claim 1 or 2, characterized in that: The water-absorbing pad is pulp paper.

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