CN210123440U

CN210123440U - Detection reagent strip, kit and detection instrument

Info

Publication number: CN210123440U
Application number: CN201920667686.8U
Authority: CN
Inventors: 周小进; 庄光磊; 缪志刚
Original assignee: Suzhou Benevolent Biomedicine Technology Co Ltd
Current assignee: Suzhou Benevolent Biomedicine Technology Co Ltd
Priority date: 2019-05-10
Filing date: 2019-05-10
Publication date: 2020-03-03
Anticipated expiration: 2029-05-10

Abstract

The utility model provides a detection reagent strip, kit and detecting instrument, detection reagent strip includes the bottom plate to and set up inspection hole and reagent strip body district on the bottom plate, reagent strip body district includes reaction hole, reagent hole, sample hole and hand (hold) district, the reagent hole is loaded with and detects required reagent; the reaction hole, the reagent hole and the sample hole are non-adsorptive high molecular material holes. The detection reagent strip provided by the utility model can be used for chemiluminescence immunodetection and immobilized enzyme immunodetection, and reagents required by detection are loaded on the detection reagent strip, so that the integration of the reagent strip is realized, the detection steps are simplified, and the pollution is avoided; the detection reagent strip is matched with a detection instrument, and can realize high-sensitivity and high-accuracy qualitative or quantitative detection and analysis on various biomolecules such as antigens, haptens, antibodies, hormones, enzymes, fatty acids, microorganisms, medicines and the like.

Description

Detection reagent strip, kit and detection instrument

Technical Field

The utility model belongs to the technical field of biological detection device, concretely relates to detect reagent strip, kit and detecting instrument.

Background

Point-of-care testing (POCT) is a testing method which can be carried out on a sampling site and can quickly obtain a testing result by using a portable analytical instrument and a matched reagent. Most mature small POCT detection methods developed in the market at present are immunochromatography, detection signals of the small POCT detection methods are immunofluorescence or colloidal gold, and the detection signals are all test strips. The immunochromatography can realize qualitative detection of a sample, but cannot complete full quantitative detection, and has low detection sensitivity and larger limitation. On the other hand, the mainstream detection instrument in the market is still a large chemiluminescence instrument with high flux, high sensitivity and high detection cost, and the detection instrument of the type is generally arranged in a large hospital or a large detection mechanism, so that field sampling, timely detection and quick result obtaining cannot be realized. In addition, most of the mainstream large-scale immunoassay instruments are based on a chemical immunoassay method, and the detection result obtained by the POCT immunochromatography method is difficult to be compared with the detection result obtained by the large-scale immunoassay instrument in the transverse direction.

However, from the market development and the trend of national advancement of staged diagnosis and treatment, it is obvious that a large-scale chemiluminescence apparatus cannot practically meet the requirements of a wide first-class hospital, a community hospital, a small-scale detection mechanism, even families and individuals in the future, and the accuracy of immunochromatography POCT detection needs to be improved. Therefore, the development of a small chemiluminescence detection instrument with small detection flux, simple operation and easily-read report is the development direction of the POCT instrument in the future, and the matched detection reagent strip is the key point for realizing the functions of the POCT instrument.

CN102147412A discloses a method and a reagent for measuring IgG of an anti-protease 3 antibody, the method is realized by a kit consisting of an analysis reagent device of enzyme-linked immunosorbent assay and a matched reagent, the analysis reagent device comprises a matrix with 8 hole sites and a handle positioned at one end of the matrix, the reagent between the holes of the reagent device is filled and discarded by a specific analysis instrument, so that a sample and the reagent react, then the value of the color of the solution after the reaction is measured, and finally the measured value is processed to obtain a detection result. The method provides an independent and single-person detection device and a matched reagent, and can realize high-efficiency and accurate detection of the anti-protease 3 antibody IgG.

CN107664635A discloses a chemiluminescent detection reagent strip, wherein both ends of the reagent strip are respectively provided with a reading hole and a sample hole, a tip head hole and a magnetic sleeve hole are sequentially arranged close to the sample hole, and the reagent strip is also provided with a reagent hole, a substrate hole and a cleaning hole; in the use, only need to await measuring the sample put into the sample downthehole, just can directly detect through test probe, in the test was accomplished the back abandonment magnetic sleeve and tip head all put back the reagent strip, only need take out the reagent strip, just can carry out next a set of sample test, effectively reduced the additional cost that brings the regular clearance of discarded object and liquid.

CN109406804A discloses a full-automatic chemiluminescence tester, which comprises a transmission rack, an incubation component, a shell breaking and sampling mechanism, a magnetic separation mechanism and a test structure, wherein the incubation component is provided with a plurality of gun heads and reagent strips, and different samples are sealed in a holding tank in the reagent strips; during testing, the shell breaking and sampling mechanism controls the gun head to sample and transfer from the reagent strip, the magnetic separation mechanism separates magnetic beads from solution, and the testing machine forms a light shielding environment and reads the light intensity of the solution to be tested; the tester can realize automatic test of a plurality of reagent strips, and simple structure, easy to maintain, and survey efficient, survey with low costsly.

However, in the prior art, the POCT instrument and the reagent strip matched with the POCT instrument still have the problems of complex structure, poor universality and the like. The POCT instrument and the matched reagent strip thereof realize signal discrimination by color development based on an enzyme linked immunosorbent assay mechanism, and have low sensitivity; the detection reagent strip based on the chemiluminescence mechanism is provided with a special magnetic sleeve hole for placing a disposable magnetic sleeve, so that the reagent strip and a corresponding detection instrument have complicated structures and large volumes, magnetic particles are lost due to multiple adsorption and transfer of the magnetic particles, and the detection sensitivity and accuracy are reduced.

Therefore, the development of a detection reagent strip which has high sensitivity, good stability, simple operation and easy result reading is an urgent problem to be solved in the field.

SUMMERY OF THE UTILITY MODEL

Aiming at the defects of the prior art, the utility model aims to provide a detection reagent strip and the application thereof, wherein, all reagents required by detection are packaged in the detection reagent strip in advance, thereby avoiding pollution and simplifying the detection steps; the reaction hole, the reagent hole and the sample hole in the reagent strip body area are non-adsorptive high polymer material holes, so that the reduction of the effective concentration of a detection reagent is effectively avoided, and the detection accuracy is improved; the detection reagent strip is matched with a detection instrument, so that high-sensitivity and high-accuracy quantitative detection of various biomolecules can be realized.

In order to achieve the purpose of the utility model, the utility model adopts the following technical proposal:

in a first aspect, the present invention provides a test reagent strip, which comprises a bottom plate, and a test hole and a reagent strip body area arranged on the bottom plate, wherein the reagent strip body area comprises a reaction hole, a reagent hole, a sample hole and a handle area, and the reagent hole is filled with a reagent required for testing;

the reaction hole, the reagent hole and the sample hole are non-adsorptive high molecular material holes.

The utility model provides a detection reagent strip is with the supporting use of POCT instrument, has considered following four aspects during the design of detection reagent strip comprehensively: simple operation, small structure, suitability for single-person small-flux detection and accuracy of detection results. The utility model provides a detect reagent strip has set up two parts in inspection hole and reagent strip body district on the bottom plate, and reagent strip body district contains reaction hole, reagent hole, sample hole and hand (hold) district again, and reaction hole, reagent hole, the sample hole in inspection hole and reagent strip body district all are independent hole structure, cuts off between each hole and does not communicate. The detection hole is an area for reading detection information by a detection instrument, the reaction hole provides a place for a sample to be detected to react with a detection reagent, all reagents required by detection are loaded in the reagent hole, the sample hole is an area for placing the sample to be detected, the handle area is a contact position of an operator and the detection reagent strip, and the detection reagent strip is provided with the relatively independent handle area, so that the pollution possibly brought by the operator in the experiment is avoided.

The utility model provides a detection reagent strip has injectd reaction hole, reagent hole, sample hole and is non-adsorptivity macromolecular material hole, is in the consideration to the testing result accuracy. The reagent strip in the prior art is formed by integral injection molding, which means that a detection hole, a reaction hole, a reagent hole and a sample hole are made of the same material, and the type and the surface adsorption property of the material are not specially selected; however, since the structural units of the polymer materials are different, the surface properties and the adsorption properties of the polymer materials are different, and if the reaction wells, the reagent wells and the sample wells are adsorptive wells (for example, polystyrene wells), the active ingredients such as proteins in the sample to be detected and the detection reagent are adsorbed in the wells, so that the effective concentration of the reagent participating in the reaction is reduced, and the accuracy of the detection result is finally affected. The utility model provides a detection reagent strip is limited to non-adsorbability macromolecular material hole with reaction hole, reagent hole, the sample hole in reagent strip body district, has effectively avoided active ingredient such as protein among detect reagent or the sample that awaits measuring to adsorb on the material surface and cause the effective concentration of the reagent of participating in the detection reaction to reduce, thereby make the detected value be closer with the true value, promoted the accuracy of testing result. The utility model discloses in to the special design in reagent strip body district reaction hole, reagent hole, sample hole realize through the independent injection moulding in inspection hole and reagent strip body district.

Preferably, the non-adsorbent polymeric material comprises acrylonitrile-butadiene-styrene copolymer, polymethyl methacrylate or polypropylene, preferably polypropylene.

Preferably, the number of the detection holes is 1;

preferably, the capacity of the detection well is 50-300 μ L, such as 55 μ L, 60 μ L, 80 μ L, 100 μ L, 120 μ L, 140 μ L, 150 μ L, 170 μ L, 190 μ L, 200 μ L, 220 μ L, 240 μ L, 250 μ L, 270 μ L or 290 μ L, and the specific values therebetween are limited to space and for the sake of brevity, and the present invention is not exhaustive of the specific values included in the range.

Preferably, the detection well and the reaction well are disposed adjacent to each other.

The inspection hole and the reagent hole of detect reagent strip are adjacent to be set up, are in order to guarantee to await measuring sample and detect reagent and mix the back and shift to the inspection hole and carry out the in-process that detects and do not take place reagent loss in the reaction hole, avoid the detection error that reagent loss brought.

Preferably, the number of the reagent wells is 1 to 10, such as 2, 3, 4, 5, 6, 7, 8 or 9, and more preferably 6 to 10;

preferably, the reagent wells have a capacity of 100-800 μ L, such as 120 μ L, 140 μ L, 150 μ L, 170 μ L, 190 μ L, 200 μ L, 220 μ L, 250 μ L, 270 μ L, 300 μ L, 320 μ L, 350 μ L, 380 μ L, 400 μ L, 430 μ L, 450 μ L, 470 μ L, 500 μ L, 520 μ L, 550 μ L, 580 μ L, 600 μ L, 630 μ L, 650 μ L, 680 μ L, 700 μ L, 720 μ L, 750 μ L or 790 μ L, and specific points between the above values are not limited to specific points included in the range for the sake of brevity and brevity.

Preferably, each of said reagent wells is loaded with a detection reagent;

preferably, the types of the detection reagent include a preservation solution for the capture antibody, a washing solution, a preservation solution for the detection antibody, a reaction substrate, a diluent, and a preservation solution for the antigen;

preferably, the preservation solution for the capture antibody is a preservation solution for magnetic particles for the capture antibody;

preferably, the detection reagent strip is a solid-phase enzyme immunoassay reagent strip, and the number of the reagent wells of the detection reagent strip is 3-10, for example, the number of the reagent wells is 3, 4, 5, 6, 7, 8, 9 or 10; the reagent holes are respectively a reagent hole for loading a preservation solution for detecting the antibody, a reagent hole for loading a reaction substrate and 1-8 reagent holes for loading a cleaning solution.

Preferably, the detection reagent strip is a chemiluminescence immunoassay reagent strip, and the number of the reagent holes of the detection reagent strip is 4-10, for example, the number of the reagent holes is 4, 5, 6, 7, 8, 9 or 10; the reagent holes are respectively a reagent hole for loading a preservation solution for capturing the antibody, a reagent hole for loading a preservation solution for detecting the antibody, a reagent hole for loading a reaction substrate and 1-7 reagent holes for loading a cleaning solution.

Preferably, the number of the reaction holes is 1-2;

preferably, the reaction well has a capacity of 400-1000 μ L, such as 410 μ L, 430 μ L, 450 μ L, 470 μ L, 500 μ L, 520 μ L, 550 μ L, 580 μ L, 600 μ L, 630 μ L, 650 μ L, 680 μ L, 700 μ L, 720 μ L, 750 μ L, 780 μ L, 800 μ L, 830 μ L, 850 μ L, 880 μ L, 900 μ L, 920 μ L, 950 μ L or 980 μ L, and specific points between the above values, limited to space and for brevity, the present invention is not exhaustive of the specific points included in the range.

Preferably, the number of the sample wells is 1;

preferably, the sample well has a capacity of 10 μ L to 200 μ L, such as 20 μ L, 30 μ L, 50 μ L, 70 μ L, 90 μ L, 100 μ L, 120 μ L, 140 μ L, 150 μ L, 170 μ L or 190 μ L, and the specific values therebetween are not exhaustive, and for brevity, the present invention does not provide any exhaustive list of the specific values included in the range.

Preferably, a bottom plate of the detection reagent strip is sequentially provided with a detection hole, a reaction hole, a reagent hole, a sample hole and a handle area from left to right;

preferably, the detection hole is a high molecular material hole;

preferably, when the detection reagent strip is used for solid-phase enzyme immunoassay, the detection holes are adsorptive high molecular material holes;

preferably, the adsorptive high molecular material is polystyrene;

preferably, the detection well has adsorbed therein a capture antibody.

The detection reagent strip provided by the utility model can be suitable for solid phase enzyme immunoassay or magnetic particle chemiluminescence immunoassay, when the detection reagent strip is the magnetic particle chemiluminescence immunoassay reagent strip, the detection hole is a high polymer material hole, and the adsorption performance of the detection hole is not specially limited; when the detection reagent strip is a solid-phase enzyme immunoassay reagent strip, the detection hole is a solid-phase carrier for capturing antibodies, and is a high-molecular material hole (such as a polystyrene hole) with adsorbability, particularly protein adsorbability, and the capture antibodies for capturing antigens or antibodies in a sample to be detected are adsorbed in the detection hole.

Preferably, the detection reagent strip is packaged by a film sealing material;

preferably, the film sealing material is a polymer film, an aluminum foil or an aluminum-plastic composite material, and more preferably an aluminum-plastic composite material.

The utility model provides an among the test reagent strip, except that the sample that awaits measuring, all the other required reagents of detection all load in advance in the reagent strip, have realized the integrated integration of reagent strip. In order to prevent the loss of the detection reagent loaded in the detection reagent strip in the transportation and storage processes, the detection reagent strip is packaged by a sealing film material. When the detection reagent strip is used, the sealing film material can be torn off or punctured by a suction head or a power extrusion device of a detection instrument.

The detection reagent strip is a myoglobin chemiluminescence immunoassay reagent strip;

preferably, the myoglobin chemiluminescence immunoassay test strip has 4-10 reagent wells, for example, 4, 5, 6, 7, 8, 9 or 10 reagent wells; the reagent holes are respectively a reagent hole for loading an antimyosin magnetic particle preservation solution, a reagent hole for loading an antimyosin antibody marked by an enzyme marker, a reagent hole for loading a reaction substrate and 1-7 reagent holes for loading a cleaning solution.

Preferably, the detection reagent strip is an amyloid protein chemiluminescence immunoassay reagent strip;

preferably, the number of the reagent wells of the amyloid chemiluminescence immunoassay reagent strip is 4-10, for example, the number of the reagent wells is 4, 5, 6, 7, 8, 9 or 10; the reagent holes are respectively a reagent hole for loading anti-amyloid magnetic particle preservation solution, a reagent hole for loading an anti-amyloid antibody marked by an enzyme marker, a reagent hole for loading a reaction substrate and 1-7 reagent holes for loading cleaning solution.

Preferably, the detection reagent strip is a myoglobin immobilized enzyme immunoassay reagent strip;

preferably, the myoglobin immobilized enzyme immunoassay reagent strip has 3-10 reagent wells, for example, 3, 4, 5, 6, 7, 8, 9 or 10 reagent wells; the reagent holes are respectively a reagent hole for loading an enzyme marker labeled anti-myoglobin antibody, a reagent hole for loading a reaction substrate and 1-8 reagent holes for loading a cleaning solution.

On the other hand, the utility model provides a kit, contain the detection reagent strip as above in the kit.

On the other hand, the utility model provides a detection instrument, the detection instrument includes incubation device and the magnetic separation device that install on the base, magnetic separation device and incubation device's upper portion is equipped with the detection reagent strip and places the district, be provided with power extrusion device, move liquid device and signal measurement device above the detection reagent strip is placed the district side by side, and with the data processing device that the signal measurement device passes through the wire and connects; the incubation device is fixedly connected with the bottom plate of the detection reagent strip through the clamping groove.

The utility model provides a detecting instrument and as above detect reagent strip mutually support, realize the quantitative determination of biomolecule, wherein detecting instrument's incubation device provides constant temperature protection for inspection hole and reaction hole, and reagent hole among the detect reagent strip, sample hole region do not set up constant temperature protection. This particular design is based on the stability of the detection reagent, since the incubation device typically provides a constant temperature of 37 ℃ for the immunoassay reaction, at which some property inactivation or denaturation of the detection reagent or some component of the sample to be tested may occur, thereby affecting the stability of the detection reagent and the accuracy of the detection result. Therefore, the incubation device in the detecting instrument only provides a constant temperature environment for the reaction hole and the detection hole of the detection reagent strip.

The working principle that the detection reagent strip and the detection instrument are mutually matched to carry out biomolecule detection is as follows: when adopting the magnetic particle chemiluminescence immunoassay method, the reagent hole of the detection reagent strip is respectively loaded with the preservation solution for capturing the antibody, the cleaning solution, the preservation solution for detecting the antibody and the reaction substrate, an operator adds a sample to be detected into the sample hole of the detection reagent strip through a liquid adding device, then the detection reagent strip is put into the detection instrument, the detection reagent strip is sequentially moved to the power extrusion device, the liquid transferring device and the corresponding working area of the detection device through the manual movement or the conveying platform in the instrument for detection, and the specific steps are as follows: a power extrusion device in the detection instrument punctures a film sealing material on the detection reagent strip, a liquid transfer device respectively sucks a sample to be detected in a sample hole and a magnetic particle preservation solution for capturing an antibody in a reagent hole, the magnetic particle preservation solutions are added into a reaction hole, and mixed reaction is carried out, wherein the operation processes of the power extrusion device and the liquid transfer device can be driven and controlled by a motor in the detection instrument; meanwhile, an incubation device in the detection instrument provides a constant temperature environment required by the reaction for the reaction hole; after the reaction in the first step is finished, a magnetic separation device in the detection instrument adsorbs magnetic particles, a liquid transfer device sucks the waste liquid and then sucks cleaning liquid in a reagent hole to clean the magnetic particles, and then the liquid transfer device adds preservation liquid for detecting antibodies into the reaction hole to perform a second step of mixed reaction; after the second mixing reaction is finished, the liquid transfer device sucks the waste liquid and then sucks the cleaning liquid in the reagent hole to clean the magnetic particles; after the cleaning is finished, the liquid transfer device sucks a reaction substrate in the reagent hole to the reaction hole, magnetic particles in the reaction hole are completely mixed with the reaction substrate, the liquid transfer device transfers the mixed liquid to the detection hole, a signal measuring device of the detection instrument reads a chemiluminescence signal in the detection hole, and then the chemiluminescence signal is converted into a concentration value of a sample to be detected through a data processing device of the instrument to obtain a detection result. And waste liquid generated in the detection process is transferred to the detection reagent strip waste hole by the liquid transfer device, and after the detection is finished, the detection reagent strip and the waste liquid are discarded along with the waste liquid.

When adopting solid-phase enzyme immunoassay, the reagent hole of detect reagent strip has loaded washing liquid, the preservative fluid and the reaction substrate of detection antibody respectively, has adsorbed the capture antibody in the inspection hole, and operating personnel adds the sample that awaits measuring to the sample hole of detect reagent strip through the liquid feeding device, then puts into the detecting instrument with the detect reagent strip, and the detect reagent strip moves in proper order to power extrusion device, move liquid device and detecting device's corresponding work area through the conveying platform in artificial removal or the instrument and detects, and concrete step is as follows: a power extrusion device in the detection instrument punctures a film sealing material on the detection reagent strip, a liquid transfer device sucks a sample to be detected in a sample hole and adds the sample into the detection hole for mixed reaction, and the operation processes of the power extrusion device and the liquid transfer device can be driven and controlled by a motor in the detection instrument; meanwhile, an incubation device in the detection instrument provides a constant temperature environment required by the reaction for the detection hole; after the first-step reaction is finished, the liquid transfer device sucks the waste liquid, then sucks the cleaning liquid in the reagent hole to clean the detection hole, and then the liquid transfer device adds the preservation liquid of the detection antibody into the detection hole to perform a second-step mixing reaction; after the second mixing reaction is finished, the liquid transfer device sucks the waste liquid and then sucks the cleaning liquid in the reagent hole to clean the detection hole; after the cleaning is finished, the pipetting device sucks the reaction substrate in the reagent hole into the detection hole, after the reaction substrate is fully mixed, a signal measuring device of the detection instrument reads a chemiluminescence signal in the detection hole, and then the chemiluminescence signal is converted into a concentration value of a sample to be detected through a data processing device of the instrument to obtain a detection result. And waste liquid generated in the detection process is transferred to the detection reagent strip waste hole by the liquid transfer device, and after the detection is finished, the detection reagent strip and the waste liquid are discarded along with the waste liquid.

Compared with the prior art, the utility model discloses following beneficial effect has:

(1) the detection reagent strip provided by the utility model pre-encapsulates other reagents required by detection except a sample to be detected into the detection reagent strip, so that the problem of adding the detection reagent is not required to be considered in the detection process, the detection procedure is simplified, and the pollution possibly brought by repeated liquid adding is avoided; after a group of detection experiments are completed, the detection reagent strip is abandoned along with the detection reagent strip, the next group of detection experiments cannot be influenced, and cross contamination cannot occur in detection.

(2) In the reagent strip for detecting provided by the utility model, the reaction hole, the reagent hole and the sample hole in the reagent strip body area are non-adsorptive high polymer material holes, and the detection hole can be flexibly selected to be a high polymer material hole or an adsorptive high polymer material hole according to different detection methods; the special design not only enlarges the application range of the detection reagent strip, but also avoids the reduction of effective concentration in the reagent by the non-adsorptive high polymer material hole, so that the detection value is closer to the true value, and the accuracy of the detection result is improved.

(3) The utility model provides a detection reagent strip is used for chemiluminescence immunodetection, and its biological carrier of catching the antibody is for having the magnetic particle of magnetic responsiveness, compares in traditional ELIAS plate, and its antibody capture ability, albumen capture efficiency, check time and sensitivity all have obvious optimization.

(4) The utility model provides a test reagent strip passes through the design in inspection hole, reagent hole and loads the selection of detect reagent, has expanded the application range of test reagent strip, can realize qualitative or quantitative detection analysis to multiple biomolecules such as antigen, hapten, antibody, hormone, enzyme, fatty acid, microorganism and medicine.

(5) The utility model provides a detect reagent strip and detecting instrument cooperation are used, have realized a plurality of functions such as the transfer of reagent, mixed reaction, constant temperature condition control, signal detection in detecting instrument, have improved the stability of detection efficiency, simple operation nature and test result.

Drawings

Fig. 1 is a schematic structural diagram of a detection reagent strip in embodiment 1 of the present invention, wherein a is a bottom plate, b is a reagent strip body region, 1 is a detection hole, 2 is a reaction hole, 3 is a reagent hole, 4 is a sample hole, and 5 is a handle region;

fig. 2 is a plan view of a detection reagent strip in embodiment 1 of the present invention, wherein 1 is a detection well, 2 is a reaction well, 3 is a reagent well, 4 is a sample well, and 5 is a handle region;

FIG. 3 is a graph showing the linear relationship between the detection value and the myoglobin concentration in example 2 of the present invention;

fig. 4 is a plan view of a detection reagent strip in embodiment 4 of the present invention, wherein 1 is a detection well, 2 is a reaction well, 3 is a reagent well, 4 is a sample well, and 5 is a handle region;

FIG. 5 is a graph showing the linear relationship between the detection value and the myoglobin concentration in example 5 of the present invention;

fig. 6 is a plan view of a detection reagent strip in embodiment 6 of the present invention, wherein 1 is a detection hole, 2 is a reaction hole, 3 is a reagent hole, 4 is a sample hole, and 5 is a handle region.

Detailed Description

The technical solution of the present invention will be further explained by the following embodiments. It should be understood by those skilled in the art that the described embodiments are merely provided to assist in understanding the present invention and should not be construed as specifically limiting the present invention.

Example 1

The embodiment provides a detection reagent strip, a schematic structural diagram of the detection reagent strip is shown in fig. 1, a plan view thereof is shown in fig. 2, and the detection reagent strip sequentially comprises from left to right: 1 represents one detection well, 2 represents two reaction wells, 3 represents nine reagent wells, 4 represents one sample well, and 5 represents a handle area. The volume of the detection well is 300. mu.L, the volume of the reaction well is 800. mu.L, the volume of the reagent well is 400. mu.L, and the volume of the sample well is 200. mu.L.

The detection hole, the reaction hole, the reagent hole and the sample hole of the detection reagent strip provided by the embodiment are polypropylene holes, the membrane sealing material is an aluminum-plastic composite material, and 9 reagent holes are sequentially loaded with magnetic particle preservation solution, cleaning solution for capturing antibodies, preservation solution, cleaning solution and reaction substrates for detecting the antibodies from left to right.

The utility model provides a detection reagent strip is used for the time measuring of biomolecule, and the reaction hole is the place that awaits measuring sample and detect reagent mix reaction, and the incubation device among the detecting instrument only provides constant temperature environment to the reaction hole position.

Example 2

In this example, 100. mu.L of a magnetic microparticle storage solution, 300. mu.L of a washing solution, 200. mu.L of an Alkaline Phosphatase (AP) -labeled anti-Myo antibody, 300. mu.L of a washing solution, 300. mu.L of a reaction substrate, and the like were sequentially loaded into 9 reagent wells from left to right in this example.

The detection method comprises the following specific steps:

(1) incubation of sample to be tested and magnetic particle preservation solution

Adding 50 mu L of sample to be detected (namely blood plasma) into a sample hole of the detection reagent strip, putting the detection reagent strip into a matched detection instrument, transferring 50 mu L of sample to be detected into a reaction hole by a liquid transfer device in the instrument, then transferring 100 mu L of anti-Myo magnetic bead preservation solution in the reagent hole into the reaction hole where the sample to be detected is located, maintaining the reaction hole in a constant temperature state of 37 ℃ by an incubation device of the instrument, and incubating for 1 hour under the condition.

(2) Cleaning of magnetic particles

After the incubation in the step (1) is finished, a magnetic separation device of the instrument adsorbs and enriches the magnetic particles in the reaction hole, and a liquid transfer device adsorbs waste liquid without the magnetic particles; then, adding the cleaning liquid in the reagent hole into the reaction hole by a liquid transfer device, closing the magnetic separation device, and blowing, beating and mixing the cleaning liquid and the magnetic particles in the reaction hole by the liquid transfer device; after 30 seconds, the magnetic separation device is started to adsorb the magnetic particles, and the liquid transfer device sucks the cleaning liquid to finish one-time cleaning; repeat the above washing step 2 times.

(3) Detecting antibody binding

After the cleaning in the step (2) is finished, transferring 200 mu L of AP labeled anti-Myo antibody in the reagent hole to a reaction hole by a liquid transferring device, and mixing the magnetic particles and the reagent by blowing and beating; the incubation apparatus was maintained at a constant temperature of 37 ℃ in the reaction well, and incubated under this condition for 1 hour.

(4) Cleaning of magnetic particles

After the incubation in the step (3) is finished, a magnetic separation device of the instrument adsorbs and enriches the magnetic particles in the reaction hole, and a liquid transfer device adsorbs the waste liquid without the magnetic particles; then, adding the cleaning liquid in the reagent hole into the reaction hole by a liquid transfer device, closing the magnetic separation device, and blowing, beating and mixing the cleaning liquid and the magnetic particles in the reaction hole by the liquid transfer device; after 30 seconds, the magnetic separation device is started to adsorb the magnetic particles, and the liquid transfer device sucks the cleaning liquid to finish one-time cleaning; repeat the above washing step 2 times.

(5) Signal detection

After the cleaning in the step (4) is finished, transferring 100 mu L of reaction substrate from the reagent hole to the reaction hole by a liquid transferring module, and uniformly blowing; and then the mixed liquid is sucked and transferred into a detection hole, a signal measuring device of the instrument collects chemiluminescent signals in the detection hole, and the chemiluminescent signals are converted into corresponding concentration values through a data processing device of the instrument, so that a myoglobin detection result is obtained.

The myoglobin standard substance is subjected to gradient dilution by using the artificial plasma to obtain a myoglobin standard solution with gradient concentration, the myoglobin standard solution is tested by the detection method, and the obtained detection values are shown in table 1:

TABLE 1

The detection values in table 1 were analyzed, and a linear relationship between the detection value and the myoglobin concentration was obtained by performing a one-dimensional linear regression analysis with the myoglobin concentration as the abscissa and the detection value as the ordinate, as shown in fig. 3, it can be seen from fig. 3 that the detection value and the myoglobin concentration have a better linear relationship,its determination coefficient R²Is 0.9997, explains the utility model provides a detect reagent strip is used for chemiluminescence immunoassay, has good detection accuracy, stability and detectivity.

Example 3

This example is an application example of the test reagent strip described in example 1 for detecting amyloid A β 1-42, and this example detects the concentration of amyloid A β 1-42 in the serum of patients with Alzheimer's disease and normal persons, respectively.

In this example, the 9 reagent wells of the test strip were loaded with 100. mu.L of the anti-A β 1-42 magnetic particle storage solution, 200. mu.L of the washing solution, 100. mu.L of the AP-labeled anti-A β 1-42 antibody, 200. mu.L of the washing solution, and 100. mu.L of the reaction substrate in this order from left to right.

The detection method comprises the following specific steps:

Taking serum samples of 16 patients with Alzheimer's disease and 16 normal persons, taking 32 detection reagent strips corresponding to the 32 serum samples, adding 50 mu L of samples to be detected (namely serum) into sample holes of the detection reagent strips, putting the detection reagent strips into a matched detection instrument, transferring 50 mu L of samples to be detected into reaction holes by a liquid transferring device in the instrument, then transferring 100 mu L of anti-A β 1-42 magnetic bead preservation solution in the reagent holes to the reaction holes where the samples to be detected are located, maintaining the reaction holes in a constant temperature state of 37 ℃ by an incubation device of the instrument, and incubating for 1 hour under the condition.

(2) Cleaning of magnetic particles

(3) Detecting antibody binding

After the completion of the washing in step (2), 100. mu.L of the AP-labeled anti-A β 1-42 antibody in the reagent well was transferred to the reaction well by the pipetting device, and the magnetic microparticles and the reagent were mixed by pipetting, and the reaction well was maintained at a constant temperature of 37 ℃ by the incubation device and incubated under these conditions for 1 hour.

(4) Cleaning of magnetic particles

(5) Signal detection

And (4) after the cleaning is finished, transferring 100 mu L of reaction substrate from the reagent hole to the reaction hole by the liquid transfer module, blowing and uniformly mixing, sucking and transferring the mixed liquid to the detection hole, collecting a chemiluminescence signal in the detection hole by a signal measuring device of the instrument, and converting the chemiluminescence signal into a corresponding concentration value by a data processing device of the instrument to obtain a detection result of the amyloid A β 1-42.

The detection of A β 1-42 in the serum of 16 patients with Alzheimer's disease and 16 normal persons was carried out according to the above method, and the obtained A β 1-42 detection values are shown in Table 2.

TABLE 2

Sample information	Detection value	Sample information	Detection value	Sample information	Detection value	Sample information	Detection value
								Patient
1	315.23	Patient 9	320.63	Normal human 1	543.6	Normal human 9	723.1
								Patient 2	190.65	Patient 10	272.09	Normal person 2	493.3	Normal human 10	324.62
Patient 3	285.94	Patient 11	239.29	Normal person 3	350.26	Normal person 11	527.55
								Patient 4	254.55	Patient 12	178.25	Normal person 4	530.16	Normal human 12	834.93
Patient 5	338.42	Patient 13	266.32	Normal human 5	270.55	Normal human 13	733.53
								Patient 6	197.76	Patient 14	331.17	Normal person 6	200.1	Normal human 14	522.96
Patient 7	291.28	Patient 15	274.65	Normal human being 7	425.08	Normal person 15	305.85
								Patient 8	176.43	Patient 16	625.4	Normal person 8	675.54	Normal person 16	239.01

The average value calculation is carried out according to the detection data in the table 2, the detection average value of the patient is 284.88, the detection average value of the normal person is 481.25, the detection average value of the patient is lower than that of the normal person, and the difference between the detection average value and the normal person is 40.81 percent, and the result accords with the result that the total amount of A β 1-42 in the body of the Alzheimer disease patient is obviously lower than that of the normal person reported in the literature, which shows that the detection data obtained by the detection reagent strip of the utility model is reliable and has referential property.

Example 4

The present embodiment provides a detection reagent strip, a plan view of the detection reagent strip is shown in fig. 4, the detection reagent strip sequentially includes from left to right: 1 represents one detection well, 2 represents one reaction well, 3 represents eight reagent wells, 4 represents one sample well, and 5 represents a handle area. The volume of the detection well was 300. mu.L, the volume of the reaction well was 600. mu.L, the volume of the reagent well was 500. mu.L, and the volume of the sample well was 150. mu.L.

In the detection reagent strip provided by the embodiment, the detection holes are polystyrene holes, the reaction holes, the reagent holes and the sample holes in the reagent strip body area are all polypropylene holes, and the film sealing material is an aluminum-plastic composite material; the capture antibody is adsorbed in the detection wells, and 8 reagent wells are sequentially loaded with a cleaning solution, a storage solution for the detection antibody, a cleaning solution, a reaction substrate from left to right.

The utility model provides a detection reagent strip is used for the time measuring of biomolecule, and the inspection hole is the place that awaits measuring sample and detect reagent mix reaction, and the incubation device among the detecting instrument only provides constant temperature environment to the inspection hole position.

Example 5

In this example, 300. mu.L of a wash solution, 200. mu.L of an AP-labeled anti-Myo antibody, 300. mu.L of a wash solution, and 100. mu.L of a reaction substrate were sequentially loaded into 8 reagent wells from left to right in order to use the test reagent strip described in example 4 for solid-phase myoglobin (Myo) detection.

The detection method comprises the following specific steps:

(1) incubation of samples to be tested

Adding 50 microliter of sample to be detected (namely blood plasma) into a sample hole of the detection reagent strip, putting the detection reagent strip into a matched detection instrument, transferring 50 microliter of sample to be detected into the detection hole by a liquid transfer device in the instrument, maintaining the detection hole in a constant temperature state of 37 ℃ by an incubation device of the instrument, and incubating for 1 hour under the condition.

(2) Cleaning of

After the incubation in the step (1) is finished, absorbing the waste liquid by a liquid transfer device; then, adding the cleaning liquid in the reagent hole into the detection hole by a liquid transfer device, and blowing and beating the cleaning liquid in the detection hole; after 30 seconds, the cleaning liquid is sucked by the liquid moving device, and one-time cleaning is completed; repeat the above washing step 2 times.

(3) Detecting antibody binding

After the washing in the step (2) is finished, transferring 200 mu L of AP labeled anti-Myo antibody in the reagent well to a detection well by a liquid transferring device; the incubation device maintained the wells at a constant temperature of 37 ℃ and incubated for 1 hour under these conditions.

(4) Cleaning of

After the incubation in the step (3) is finished, absorbing the waste liquid by a liquid transfer device; then, adding the cleaning liquid in the reagent hole into the detection hole by a liquid transfer device, and blowing and beating the cleaning liquid in the detection hole; after 30 seconds, the cleaning liquid is sucked by the liquid moving device, and one-time cleaning is completed; repeat the above washing step 2 times.

(5) Signal detection

And (4) after the cleaning in the step (4) is finished, transferring 100 mu L of reaction substrate from the reagent hole to the detection hole by the liquid transferring module, then collecting a chemiluminescence signal in the detection hole by a signal measuring device of the instrument, and converting the chemiluminescence signal into a corresponding concentration value by a data processing device of the instrument to obtain a myoglobin detection result.

The myoglobin standard substance was diluted with artificial plasma in a gradient manner to obtain a myoglobin standard solution with a gradient concentration, and the myoglobin standard solution was tested by the above-mentioned test method, and the obtained test values are shown in table 3:

TABLE 3

The detection values in table 3 were analyzed, and a linear relationship between the detection value and the myoglobin concentration was obtained by subjecting the myoglobin concentration as abscissa and the detection value as ordinate to a linear regression analysis, as shown in fig. 5, and it can be seen from fig. 5 that the detection value and the myoglobin concentration have a good linear relationship, and the determination coefficient R thereof²0.9991, it is shown that the test reagent strip provided by the utility model is used for solid-phase enzyme immunoassay, and has good detection accuracy, stability and detection sensitivity.

Example 6

The present embodiment provides a detection reagent strip, a plan view of the detection reagent strip is shown in fig. 6, the detection reagent strip sequentially includes from left to right: 1 represents one detection well, 2 represents two reaction wells, 3 represents six reagent wells, 4 represents one sample well, and 5 represents a handle area. The volume of the detection well was 200. mu.L, the volume of the reaction well was 700. mu.L, the volume of the reagent well was 700. mu.L, and the volume of the sample well was 100. mu.L.

The detection hole, the reaction hole, the reagent hole and the sample hole in the detection reagent strip provided by the embodiment are all polypropylene holes, and the film sealing material is an aluminum-plastic composite material; the 6 reagent wells were loaded with a magnetic particle storage solution for capturing an antibody, a storage solution for detecting an antibody, a washing solution, and a reaction substrate in this order from left to right.

Example 7

This example is an application of the test reagent strip described in example 6 to the detection of amyloid A β 1-42, and this example separately detects the concentration of amyloid A β 1-42 in the serum of patients with Alzheimer's disease and normal persons.

In this example, 6 reagent wells of the test strip were loaded with 100. mu.L of the anti-A β 1-42 magnetic microparticle storage solution, 100. mu.L of the AP-labeled anti-A β 1-42 antibody, 200. mu.L of the washing solution, and 100. mu.L of the reaction substrate in this order from left to right.

The detection method comprises the following specific steps:

(1) incubation of sample to be detected, magnetic particle preservation solution and AP-labeled anti-A β 1-42 antibody

Taking serum samples of 16 patients with Alzheimer's disease and 16 normal persons, taking 32 detection reagent strips corresponding to the 32 serum samples, adding 50 mu L of samples to be detected (namely serum) into sample holes of the detection reagent strips, putting the detection reagent strips into a matched detection instrument, transferring 50 mu L of samples to be detected into reaction holes by a liquid transferring device in the instrument, then transferring 100 mu L of anti-A β 1-42 magnetic bead preservation solution and 100 mu L of AP labeled anti-A β 1-42 antibody in the reagent holes to the reaction holes where the samples to be detected are positioned, maintaining the reaction holes in a constant temperature state of 37 ℃ by an incubation device of the instrument, and incubating for 1 hour under the condition.

(2) Cleaning of magnetic particles

(3) Signal detection

And (3) after the cleaning in the step (2) is finished, transferring 100 mu L of reaction substrate from the reagent hole to the reaction hole by the liquid transfer module, blowing and uniformly mixing, sucking and transferring the mixed liquid to the detection hole, collecting a chemiluminescence signal in the detection hole by a signal measuring device of the instrument, and converting the chemiluminescence signal into a corresponding concentration value by a data processing device of the instrument to obtain a detection result of the amyloid A β 1-42.

TABLE 4

Sample information	Detection value	Sample information	Detection value	Sample information	Detection value	Sample information	Detection value
								Patient
1	427.14	Patient 9	423.26	Normal human 1	646.23	Normal human 9	825.73
								Patient 2	278.63	Patient 10	386.42	Normal person 2	595.93	Normal human 10	1027.25
Patient 3	239.57	Patient 11	419.01	Normal person 3	452.62	Normal person 11	630.18
								Patient 4	357.18	Patient 12	280.88	Normal person 4	718.79	Normal human 12	937.56
Patient 5	541.05	Patient 13	368.52	Normal human 5	573.34	Normal human 13	836.16
								Patient 6	363.39	Patient 14	433.8	Normal person 6	302.73	Normal human 14	625.59
Patient 7	393.91	Patient 15	377.28	Normal human being 7	527.71	Normal person 15	408.48
								Patient 8	283.06	Patient 16	728.03	Normal person 8	778.17	Normal person 16	1163.72

The average value calculation is carried out according to the detection data in the table 4, the detection average value of the patient is 393.82, the detection average value of the normal person is 690.63, the detection average value of the patient is lower than that of the normal person, and the difference between the detection average value and the normal person is 43 percent, which is consistent with the result that the total amount of A β 1-42 in the body of the Alzheimer disease patient is obviously lower than that of the normal person reported in the literature, and the detection data obtained by the detection reagent strip of the utility model is reliable and has referential property.

The applicant states that the present invention provides a reagent strip and a process method using the same through the above embodiments, but the present invention is not limited to the above process steps, i.e. the present invention must rely on the above process steps to implement. It should be clear to those skilled in the art that any improvement of the present invention is to the equivalent replacement of the selected raw materials, the addition of auxiliary components, the selection of specific modes, etc., all fall within the protection scope and disclosure scope of the present invention.

Claims

1. A detection reagent strip is characterized by comprising a bottom plate, a detection hole and a reagent strip body area, wherein the detection hole and the reagent strip body area are arranged on the bottom plate;

2. The test strip of claim 1, wherein the non-absorbent polymeric material comprises acrylonitrile-butadiene-styrene copolymer, polymethyl methacrylate, or polypropylene.

3. The test strip of claim 2, wherein the non-absorbent polymeric material is polypropylene.

4. The test reagent strip of claim 1, wherein the number of test wells is 1.

5. The test reagent strip according to claim 1, wherein the capacity of the test well is 50 to 300. mu.L.

6. The test reagent strip of claim 1, wherein the test well and the reaction well are disposed adjacent to each other.

7. The test reagent strip according to claim 1, wherein the number of the reagent wells is 1 to 10.

8. The test reagent strip according to claim 7, wherein the number of the reagent wells is 6 to 10.

9. The test reagent strip according to claim 1, wherein the reagent well has a capacity of 100 to 800. mu.L.

10. The test reagent strip of claim 7, wherein each of the reagent wells is loaded with one test reagent.

11. The test reagent strip according to claim 10, wherein the types of the test reagents include a preservation solution for capture antibodies, a washing solution, a preservation solution for test antibodies, a reaction substrate, a diluting solution, and a preservation solution for antigens.

12. The test reagent strip according to claim 11, wherein the preservation solution for the capture antibody is a preservation solution for magnetic microparticles for the capture antibody.

13. The detection reagent strip according to claim 7, wherein the detection reagent strip is a reagent strip for immobilized enzyme immunoassay, the number of the reagent wells of the detection reagent strip is 3 to 10, and the reagent wells are a reagent well for holding a storage solution for a detection antibody, a reagent well for holding a reaction substrate, and 1 to 8 reagent wells for holding a washing solution, respectively.

14. The test reagent strip according to claim 7, wherein the test reagent strip is a chemiluminescent immunoassay reagent strip, the number of the reagent wells of the test reagent strip is 4 to 10, and the reagent wells are a reagent well containing a preservation solution for a capture antibody, a reagent well containing a preservation solution for a test antibody, a reagent well containing a reaction substrate, and 1 to 7 reagent wells containing a washing solution.

15. The test reagent strip according to claim 1, wherein the number of the reaction wells is 1 to 2.

16. The test reagent strip according to claim 1, wherein the reaction well has a capacity of 400 to 1000. mu.L.

17. The test reagent strip of claim 1, wherein the number of sample wells is 1.

18. The test reagent strip of claim 1, wherein the sample well has a capacity of 10 to 200 μ L.

19. The test reagent strip of claim 1, wherein the bottom plate of the test reagent strip is provided with a detection hole, a reaction hole, a reagent hole, a sample hole and a handle area in sequence from left to right.

20. The test reagent strip of claim 1, wherein the test wells are polymer wells.

21. The test reagent strip of claim 1, wherein the test reagent strip is a immobilized enzyme immunoassay reagent strip, and the test wells are adsorptive polymer material wells.

22. The test strip of claim 21, wherein the absorbent polymer material is polystyrene.

23. The test reagent strip of claim 21, wherein the detection well has a capture antibody adsorbed therein.

24. The test reagent strip of claim 1, wherein the test reagent strip is encapsulated with a film sealing material.

25. The strip according to claim 24, wherein the sealing material is a polymer film, an aluminum foil or an aluminum-plastic composite material.

26. The test reagent strip of claim 25, wherein the sealing film material is an aluminum plastic composite material.

27. The test reagent strip of claim 1, wherein the test reagent strip is a myoglobin chemiluminescent immunoassay test reagent strip.

28. The strip according to claim 27, wherein the number of the reagent wells of the myoglobin chemiluminescence immunoassay reagent strip is 4 to 10, and the reagent wells are a reagent well for containing a preservation solution for magnetic microparticles of anti-myoglobin, a reagent well for containing an anti-myoglobin antibody labeled with an enzyme label, a reagent well for containing a reaction substrate, and 1 to 7 reagent wells for containing a washing solution.

29. The test reagent strip of claim 1, wherein the test reagent strip is an amyloid protein chemiluminescence immunoassay test reagent strip.

30. The detection reagent strip according to claim 29, wherein the number of the reagent wells of the amyloid chemiluminescence immunoassay reagent strip is 4 to 10, and the reagent wells are a reagent well for loading an anti-amyloid magnetic particle preservation solution, a reagent well for loading an anti-amyloid antibody labeled with an enzyme label, a reagent well for loading a reaction substrate, and 1 to 7 reagent wells for loading a washing solution.

31. The test reagent strip of claim 1, wherein the test reagent strip is a myoglobin immobilized enzyme immunoassay reagent strip.

32. The strip according to claim 31, wherein the number of the reagent wells of the myoglobin immobilized enzyme immunoassay reagent strip is 3 to 10, and the reagent wells are a reagent well for loading an anti-myoglobin antibody labeled with an enzyme label, a reagent well for loading a reaction substrate, and 1 to 8 reagent wells for loading a washing solution.

33. A kit comprising a test strip according to any one of claims 1 to 32.

34. The detection instrument is characterized by comprising an incubation device and a magnetic separation device which are arranged on a base, wherein the upper parts of the magnetic separation device and the incubation device are provided with a detection reagent strip placing area, and a power extrusion device, a liquid transfer device, a signal measurement device and a data processing device which is connected with the signal measurement device through a lead are arranged above the detection reagent strip placing area side by side; the incubation device is fixedly connected with the bottom plate of the detection reagent strip of any one of claims 1-32 through a clamping groove.