Reagent strip for detecting orthostereotyped blood types, preparation method of reagent strip and reagent card
阅读说明：本技术 一种正定型血型检测试剂条、试剂条制备方法及试剂卡 (Reagent strip for detecting orthostereotyped blood types, preparation method of reagent strip and reagent card ) 是由 曹大烨 张捷李 雷洋 白佳委 胡啸 黄认训 于 2019-11-29 设计创作，主要内容包括：本发明公开了一种正定型血型检测试剂条、试剂条制备方法及试剂卡,所述试剂条包括渗滤层和吸水层,所述渗滤层设于所述吸水层的上方并覆盖加样孔的对应区域,其特征在于,所述渗滤层从上到下依次包括：抗体垫、第一玻纤垫、第二玻纤垫,所述吸水层设于所述第二玻纤垫之下,所述抗体垫是采用玻纤垫包被对应血型的抗体总成液制成；试剂卡与试剂条的结构相匹配；不仅检测结果更准确,而且结构更紧凑,布局更合理。(The invention discloses a reagent strip for detecting orthostereotyped blood types, a preparation method of the reagent strip and a reagent card, wherein the reagent strip comprises a percolation layer and a water absorption layer, the percolation layer is arranged above the water absorption layer and covers the corresponding area of a sample adding hole, and the reagent strip is characterized in that the percolation layer sequentially comprises the following components from top to bottom: the water absorption layer is arranged below the second glass fiber pad, and the antibody pad is prepared by coating antibody assembly liquid of a corresponding blood type by the glass fiber pad; the structure of the reagent card is matched with that of the reagent strip; not only the testing result is more accurate, and the structure is compacter moreover, and the overall arrangement is more reasonable.)
1. The utility model provides an orthostereotype blood group test reagent strip, includes the filtration layer and the layer that absorbs water, the filtration layer is located the top on layer that absorbs water and cover the corresponding region of application of sample hole, its characterized in that, filtration layer from the top down includes in proper order: the anti body fills up, the glass fiber pad of first glass, the glass fiber pad of second, the layer that absorbs water is located under the glass fiber pad of second, the anti body fills up and is made by the antibody assembly liquid that adopts the glass fiber pad to wrap corresponding blood group.
2. The kit of claim 1, wherein: the water absorption layer comprises a first water absorption layer and a second water absorption layer, the first water absorption layer is arranged on two sides of the percolation layer, and the second water absorption layer is fixedly bonded below the second glass fiber mat; the first water-absorbing layer is provided above the second water-absorbing layer.
3. The kit of claim 2, wherein: the lengths of the antibody pad, the first glass fiber pad and the second glass fiber pad are the same, and the antibody pad and the water absorption layer are connected and fixed through a fixing layer; wherein the fixing layer adopts transparent adhesive tape, and the water absorbing layer adopts absorbent paper; the fixing layers comprise two fixing layers which are respectively arranged on two sides of the antibody pad, one end of each fixing layer is connected with the antibody pad, and the other end of each fixing layer is connected with the second water absorbing layer; the first water absorption layer and the percolation layer are arranged at intervals along the length direction of the second water absorption layer.
4. The kit of claim 2, wherein: the sizes of the antibody pad, the first glass fiber pad and the second glass fiber pad are respectively 7mm long and 5mm wide; the size of the fixed layer is 12mm in length and 5mm in width; the first water absorption layer is 19mm long and 5mm wide; the second water absorption layer has the size of 50mm in length and 5mm in width.
5. A method for preparing the reagent strip for testing orthotyping blood group according to any one of claims 1 to 4, comprising the steps of:
mixing the corresponding blood type antibody with an antibody preservation solution to prepare an antibody assembly solution;
spreading the glass fiber pad to coat the antibody assembly liquid, and drying to obtain an antibody pad;
fixing a second glass fiber mat on the second water absorption layer, and sequentially placing a first glass fiber mat and an antibody mat on the second glass fiber mat;
and fixing the edges of the two sides of the antibody pad and the second water absorption layer to obtain the reagent strip for detecting the orthostereotyped blood types.
6. The method of claim 5, wherein: mixing the blood group antibody with the antibody preservation solution in a volume ratio of 1: 5; the drying temperature of the antibody pad is 37 ℃, and the drying time is 4-5 hours.
7. The method of claim 5, wherein: the formula of the antibody preservation solution comprises: 1000ml of ultrapure water, 2ml-4ml of Tris, 6g g of BSA, 5g-15g of sucrose, 0.5g of PVP0.34 ml of HCl, 42.2 g-2.6g of Na2HPO42, 0.65g-0.9g of NaCl, 3.6g-4.2g of casein and 0.1% of preservative K.
8. A reagent card comprising the reagent strip for testing orthotyping blood group of any one of claims 1 to 4, which is prepared by the preparation method of any one of claims 5 to 7.
9. The reagent card of claim 8, wherein the reagent card comprises an upper cover and a base, the upper cover is provided with a sample hole, the base is provided with a clamping groove for mounting the reagent strip, and the sample hole corresponds to the middle position of an antibody pad of the reagent strip; the upper cover is provided with one of a fixed column or a fixed sleeve, the base is provided with the other of the fixed column or the fixed sleeve at a corresponding position, and the upper cover and the base are inserted and fixed through the fixed column and the fixed sleeve.
10. The reagent card of claim 9, wherein the card slot comprises a positioning slot extending along a length of the reagent strip, a positioning rib extending along the length of the reagent strip, end supports corresponding to two ends of the positioning slot, and a middle support corresponding to a middle of the positioning slot, wherein the positioning rib is located inside the positioning slot, and the end supports and the middle support are located outside the positioning slot; and the inner side of the upper cover is provided with a positioning strip for positioning the antibody pad corresponding to the position of the antibody pad, and positioning columns for positioning the first water absorption layers corresponding to the positions of the first water absorption layers on the two sides of the antibody pad.
Technical Field
The invention relates to a reagent strip for detecting orthostereotyped blood types, a preparation method of the reagent strip and a reagent card.
Background
ABO blood group testing is mainly used for: clinical blood transfusion, selection of donors with corresponding ABO blood types during organ transplantation such as skin and kidney transplantation, analysis of etiology of infertility and neonatal hemolysis, paternity test, and the like. Blood type is not only important in blood transfusion, but also has application value in anthropology, genetics, forensic science, transplantation immunity, disease resistance (or susceptibility), etc., before blood transfusion, the blood types of patients (recipients) and transfusion persons (donors) must be checked, and cross matching tests are performed. In clinical medicine, besides blood transfusion and transplantation, blood type knowledge and related technologies are required for examining specific antibodies of hemolytic disease of newborn and autoimmune hemolytic anemia.
The mainstream detection reagents for detecting blood group antigens in the market mainly comprise: blood type detection reagent, microcolumn gel detection reagent and blood type solid phase chromatography are used for detecting several types; wherein, blood group detection reagent and microcolumn gel detection reagent need tedious operation, and have certain qualification requirements to the operating personnel, still need the centrifugal equipment in the reaction, very easy to cause the blood group to appraise the mistake, even the serious transfusion accident appears; the traditional blood type solid phase chromatography needs to evaluate a detection result according to a detection line (T line) and a quality control line (C line), so that the operation is complex, and the accuracy and the stability of the detection result are poor when the detection line is fuzzy or the test paper is invalid.
Disclosure of Invention
The invention provides a reagent strip for detecting orthostereotyped blood types, a preparation method of the reagent strip and a reagent card for solving the problems, and the reagent strip has a more compact structure and higher accuracy.
In order to achieve the purpose, the invention adopts the technical scheme that:
one of the objects of the present invention is: the utility model provides an orthostereotype blood group testing reagent strip, include the filtration layer and absorb water the layer, the filtration layer is located the top on layer that absorbs water and cover the corresponding region of application of sample hole, filtration layer from the top down includes in proper order: the anti body fills up, the glass fiber pad of first glass, the glass fiber pad of second, the layer that absorbs water is located under the glass fiber pad of second, the anti body fills up and is made by the antibody assembly liquid that adopts the glass fiber pad to wrap corresponding blood group.
Preferably, the water absorption layer comprises a first water absorption layer and a second water absorption layer, the first water absorption layer is arranged on two sides of the percolation layer, and the second water absorption layer is fixedly bonded below the second glass fiber mat; the first water-absorbing layer is provided above the second water-absorbing layer.
Preferably, the lengths of the antibody pad, the first glass fiber pad and the second glass fiber pad are the same, and the antibody pad and the water absorption layer are connected and fixed through a fixing layer; wherein the fixing layer adopts transparent adhesive tape, and the water absorbing layer adopts absorbent paper; the fixing layers comprise two fixing layers which are respectively arranged on two sides of the antibody pad, one end of each fixing layer is connected with the antibody pad, and the other end of each fixing layer is connected with the second water absorbing layer; the first water absorption layer and the percolation layer are arranged at intervals along the length direction of the second water absorption layer.
Preferably, the antibody pad, the first glass fiber pad and the second glass fiber pad have the dimensions of 7mm in length and 5mm in width respectively; the size of the fixed layer is 12mm in length and 5mm in width; the first water absorption layer is 19mm long and 5mm wide; the second water absorption layer has the size of 50mm in length and 5mm in width.
The second purpose of the invention is: the preparation method of the reagent strip for detecting the orthostereotyped blood group comprises the following steps:
mixing the corresponding blood type antibody with an antibody preservation solution to prepare an antibody assembly solution;
spreading the glass fiber pad to coat the antibody assembly liquid, and drying to obtain an antibody pad;
fixing a second glass fiber mat on the second water absorption layer, and sequentially placing a first glass fiber mat and an antibody mat on the second glass fiber mat;
and fixing the edges of the two sides of the antibody pad and the second water absorption layer to obtain the reagent strip for detecting the orthostereotyped blood types.
Preferably, the blood group antibody and the antibody preservation solution are mixed in a volume ratio of 1: 5; the drying temperature of the antibody pad is 37 ℃, and the drying time is 4-5 hours.
Preferably, the antibody preservative fluid formula comprises: 1000ml of ultrapure water, 2ml-4ml of Tris, 6g g of BSA, 5g-15g of sucrose, 0.5g of PVP0.34 ml of HCl, 42.2 g-2.6g of Na2HPO42, 0.65g-0.9g of NaCl, 3.6g-4.2g of casein and 0.1% of preservative K.
The third purpose of the invention is that: providing a reagent card comprising the reagent strip for detecting the orthostereotyped blood group, wherein the reagent strip is prepared by the preparation method.
Preferably, the reagent card comprises an upper cover and a base, the upper cover is provided with a sample adding hole, the base is provided with a clamping groove for assembling the reagent strip, and the sample adding hole corresponds to the middle position of the antibody pad of the reagent strip; the upper cover is provided with one of a fixed column or a fixed sleeve, the base is provided with the other of the fixed column or the fixed sleeve at a corresponding position, and the upper cover and the base are inserted and fixed through the fixed column and the fixed sleeve.
Preferably, the clamping groove is provided with a positioning groove extending along the length direction of the reagent strip, positioning ribs extending along the length direction of the reagent strip, end supporting frames corresponding to two ends of the positioning groove, and a middle supporting frame corresponding to the middle of the positioning groove, wherein the positioning ribs are located in the positioning groove, and the end supporting frames and the middle supporting frame are located outside the positioning groove; and the inner side of the upper cover is provided with a positioning strip for positioning the antibody pad corresponding to the position of the antibody pad, and positioning columns for positioning the first water absorption layers corresponding to the positions of the first water absorption layers on the two sides of the antibody pad.
The invention has the beneficial effects that:
(1) the percolation layer of the reagent strip is formed by longitudinally and sequentially laminating one layer of antibody pad and two layers of glass fiber pads, so that the detection result is more accurate, the structure is more compact, and the layout is more reasonable;
(2) the bottom and two sides of the percolation layer are respectively provided with the water absorption layers, so that the water absorption performance is better;
(3) the second glass fiber mat is bonded and fixed with the second water absorption layer, and the antibody mat is bonded and fixed with the second water absorption layer through the fixing layer, so that the structure is more reliable;
(4) the reagent strip is prepared on the basis of combining the structure of the reagent strip, so that the process is simplified, and the performance is more stable;
(5) the formula of the antibody preservation solution can obviously improve the activity of the preserved protein, improve the stability of the protein after being coated on the glass fiber mat and provide the preservation period of the protein in a normal temperature environment
(6) The structure of the reagent card is more reliable in positioning and better in matching effect with the structure of the reagent strip.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the invention and not to limit the invention. In the drawings:
FIG. 1 is a schematic top view of a reagent strip for testing orthotyping blood type according to the present invention;
FIG. 2 is a schematic view of an exploded structure of the reagent strip for testing orthostereotyped blood types according to the present invention;
FIG. 3 is a schematic view of a stacking method of the reagent strips for orthotyping blood group testing according to the present invention;
FIG. 4 is a schematic perspective view of an orthostereotyped blood group testing reagent card according to one embodiment of the present invention;
FIG. 5 is a schematic view of an upper cover structure of the blood type testing reagent card according to an embodiment of the present invention;
FIG. 6 is a schematic view of a base structure of a blood type testing reagent card according to an embodiment of the present invention;
FIG. 7 is a schematic perspective view of an orthostereotyped blood group testing reagent card according to another embodiment of the present invention;
FIG. 8 is a schematic view of the upper cover structure of the blood group testing card according to another embodiment of the present invention;
FIG. 9 is a schematic view of a base structure of a blood type testing reagent card according to another embodiment of the present invention;
in the figure:
A. a percolation layer; a1, antibody pad; a2, a first glass fiber mat; a3, a second glass fiber mat; L-A, length of the percolation layer;
B. a fixing layer (transparent adhesive tape); L-B, fixed layer length;
C. a first water-absorbing layer (two-sided water-absorbing paper); L-C, length of the first water absorption layer;
D. a second absorbent layer (bottom absorbent paper); L-D, length of the second water-absorbing layer;
10. an upper cover; 11. a sample application hole; 12. fixing a column; 13. a positioning bar; 14. a positioning column;
20. a base; 21. fixing a sleeve; 22. end supports (C-shaped or C-like); 23. a first middle support frame (T-shaped); 24. a second middle support frame (I-shaped); 25. positioning ribs; 26. and (6) positioning a groove.
Detailed Description
In order to make the technical problems, technical solutions and advantageous effects to be solved by the present invention clearer and clearer, the present invention is further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The surface of human erythrocytes contains blood group antigen substances, while serum contains specific antibodies. When a red blood cell containing a blood group antigen encounters an antibody corresponding to it, a reaction occurs which causes the red blood cell to clump into a mass, in short, an antigen-antibody reaction. The ABO blood types can be classified into A, B, AB and O types. Wherein, the surface of the erythrocyte of the human of the blood group A contains the antigen A; and the red blood cells of the human with the blood type B contain B antigens; two antigens, A and B, are present on AB type human erythrocytes; neither antigen is present on the surface of human erythrocytes from blood type O.
The invention combines the antigen-antibody immunoreaction with the solid phase percolation method, and integrates to obtain a comprehensive detection method which is pertinently applied to blood type positive typing detection and RhD typing detection, so that the detection result is more accurate and reliable, and the structures of the detection reagent strip and the detection reagent card are more simplified.
First embodiment (reagent strip structure)
The reagent strip of the embodiment is a design which is established on the basis of a solid-phase percolation method and is suitable for the requirements of blood type positive typing detection sensitivity and specificity, the design comprises the arrangement of the relative physical positions of all parts of a reagent card and the construction sequence of all parts of the reagent card, and the design specifically comprises the following steps:
as shown in fig. 1 to 3, the present invention provides a reagent strip for detecting orthostereotyped blood types, which comprises a permeation layer a and a water absorption layer, wherein the permeation layer a is disposed above the water absorption layer and covers a corresponding area of a
In this embodiment, the water-absorbing layer includes a first water-absorbing layer C and a second water-absorbing layer D, the first water-absorbing layer C is disposed on two sides of the percolation layer a, the second water-absorbing layer D is adhesively fixed under the second glass fiber mat A3, and preferably, the second water-absorbing layer D and the second glass fiber mat A3 are adhesively fixed by using a double-sided adhesive tape; the first water absorption layer C is disposed above the second water absorption layer D, and the first water absorption layer C and the second water absorption layer D may be directly stacked, or may be fixed by a double-sided tape, or may be compressed by a case of a reagent card.
The lengths of the antibody pad A1, the first glass fiber pad A2 and the second glass fiber pad A3 are the same, and the antibody pad A1 and the water absorption layer are fixedly connected through a fixing layer B; wherein the fixing layer B adopts transparent adhesive tape, and the water absorbing layer adopts absorbent paper; the fixing layer B comprises two fixing layers which are respectively arranged on two sides of the antibody pad A1, one end of the fixing layer B is connected with the antibody pad A1, and the other end of the fixing layer B is connected with the second water absorption layer D; the first water absorption layer C and the percolation layer A are arranged at intervals along the length direction of the second water absorption layer D.
Specifically, the antibody pad A1, the first fiberglass pad A2 and the second fiberglass pad A3 have the dimensions of 7mm in length (L-A) and 5mm in width, respectively; the size of the fixed layer B is 12mm in length (L-B) and 5mm in width; the first water absorption layer C is 19mm long (L-C) and 5mm wide; the second water absorption layer D has the size of 50mm in length (L-D) and 5mm in width; the fixing layer B covers the two edges of the antibody pad A1 with the width of 2mm at the edge for pasting and fixing combination. The sizes of the first water absorption layer C and the second water absorption layer D can be correspondingly changed according to the change of the length and the width of the reagent strip, and the sizes of the first water absorption layer C and the second water absorption layer D can be correspondingly changed according to the different requirements of water absorption performance.
Wherein the antibody pad and the first glass fiber pad A2 are 1200-15 of Xiamen Boon biological products, and the second glass fiber pad A3 is DL-42 of Shanghai gold standard products; the fixing layer B is made of transparent adhesive tape, the first water absorption layer C is made of Shanghai gold mark CH-37 water absorption paper, and the second water absorption layer D is made of any one of Shanghai gold mark SX-42 or Auger XJ-420, XJ-440 and XJ-520 water absorption paper; the antibody pad A1 is prepared by coating antibody assembly liquid of corresponding blood type with Shanghai gold label 8975 glass fiber pad; the blood group antibodies are the corresponding blood group antibodies purchased from millipore.
Second embodiment (method for preparing reagent strip)
The present embodiment provides the method for preparing the reagent strip for testing orthotyping blood group according to any one of the first embodiment, which includes the following steps:
(1) mixing the corresponding blood type antibody with an antibody preservation solution to prepare an antibody assembly solution;
(2) spreading the glass fiber pad with the antibody assembly liquid, and drying to obtain an antibody pad A1;
wherein the drying temperature of the antibody pad A1 is 37 ℃, and the drying time is 4-5 hours.
(3) Fixing the second glass fiber pad A3 on the second water-absorbing layer D, and sequentially placing the first glass fiber pad A2 and the antibody pad A1 on the second glass fiber pad A3;
(4) and fixing the edges of two sides of the antibody pad A1 and the second water absorption layer D to obtain the reagent strip for detecting the orthostereotyped blood types.
The formula of the antibody preservation solution comprises: 1000ml of ultrapure water, 2ml-4ml of Tris, 6g g of BSA, 5g-15g of sucrose, 0.5g of PVP0.34 ml of HCl, 42.2 g-2.6g of Na2HPO42, 0.65g-0.9g of NaCl, 3.6g-4.2g of casein and 0.1% of preservative K. And mixing the blood group antibody with the antibody preservation solution in a volume ratio of 1: 5.
One of the preferable formulas of the antibody preservation solution is as follows: 1000ml of ultrapure water, 2ml of Tris, 6g of BSA, 5g of sucrose, 0.5g of PVP0, 1.34ml of HCl, 42.2g of Na2HPO0.65 g of NaCl, 3.6g of casein and 0.1% of preservative K;
the second preferable formula of the antibody preservation solution is as follows: 1000ml of ultrapure water, 2.8ml of Tris, 6g g of BSA, 8g of sucrose, 0.5g of PVP0, 1.34ml of HCl, 0.75g of Na2HPO42, 3.8g of NaCl, 0.1% of casein and 0.1% of preservative K;
the third preferable formula of the antibody preservation solution is as follows: 1000ml of ultrapure water, Tris3.2ml, BSA6g, 10g of cane sugar, 0.5g of PVP0, 1.34ml of HCl, 42.4g of Na2HPO42, 0.85g of NaCl, 4g of casein and 0.1% of preservative K;
the preferable formula of the antibody preservative fluid is four: 1000ml of ultrapure water, 4ml of Tris, 6g of BSA, 15g of sucrose, 0.5g of PVP0, 1.34ml of HCl, 42.6 g of Na2HPO42, 0.9g of NaCl, 4.2g of casein and 0.1% of preservative K.
The antibody preservation solution adopting the formula can obviously improve the activity of the preserved protein, improve the stability of the protein after being coated on the glass fiber mat and provide the preservation period of the protein in a normal temperature environment.
Third embodiment (Single-well reagent card Structure)
The present embodiment provides a reagent card, including the reagent strip for testing orthotyping blood group of any one of the first embodiment, wherein the reagent strip is manufactured by the manufacturing method of any one of the second embodiment.
As shown in fig. 4 to 6, in the present embodiment, the reagent card includes an
The
The reagent strip positioning structure comprises a positioning groove 26 extending along the length direction of the reagent strip, positioning ribs 25 extending along the length direction of the reagent strip, end supporting frames 22 corresponding to two ends of the positioning groove 26, and a middle supporting frame corresponding to the middle of the positioning groove 26, wherein the positioning ribs 25 are positioned in the positioning groove 26, and the end supporting frames 22 and the middle supporting frame are positioned outside the positioning groove 26; a positioning strip 13 for positioning the antibody pad a1 is arranged at a position corresponding to the antibody pad a1 inside the upper cover 10, and positioning columns 14 for positioning the first water absorption layer C corresponding to the positions of the first water absorption layers C on both sides of the antibody pad a 1; the middle support frame further comprises more than one first middle support frame 23 and more than one second middle support frame 24, and the middle support frames can be arranged according to the length of the reagent card; the first middle supporting frame 23 and the second middle supporting frame 24 can be in a shape of a straight line or a T shape and are symmetrically arranged at the left side and the right side of the positioning groove 26; the end supports 22 are preferably of a C-shaped or C-like design.
The application process of the reagent card of the present embodiment is briefly described as follows:
firstly, 10ul of whole blood sample is added into the
Fourth embodiment (porous reagent card structure)
As shown in fig. 7 to 9, the main difference between the present embodiment and the third embodiment is: more than two
While the foregoing description shows and describes the preferred embodiments of the present invention, it is to be understood that the invention is not limited to the forms disclosed herein, but is not to be construed as excluding other embodiments and is capable of use in various other combinations, modifications, and environments and is capable of changes within the scope of the inventive concept as described herein, commensurate with the above teachings, or the skill or knowledge of the relevant art. And that modifications and variations may be effected by those skilled in the art without departing from the spirit and scope of the invention as defined by the appended claims.
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