CN114942328A

CN114942328A - Rapid detection structure for blood insulin level and application thereof

Info

Publication number: CN114942328A
Application number: CN202210688362.9A
Authority: CN
Inventors: 刘默文
Original assignee: Jiaxing Kangyuan Ketai Technology Development Co ltd
Current assignee: Jiaxing Kangyuan Ketai Technology Development Co ltd
Priority date: 2022-06-17
Filing date: 2022-06-17
Publication date: 2022-08-26

Abstract

The invention discloses a rapid detection structure for blood insulin level and application thereof, which is a combined set of a capillary siphon sampling structure, a detection liquid loading structure and a chromatography detection structure, wherein the capillary siphon sampling structure is a capillary siphon quantitative tubular structure with openings at two ends, one end of the capillary siphon quantitative tubular structure can be directly used for quantitative collection of a blood sample, a first insulin-resistant antibody marked by an indicator is coated on a marker binding pad in a detection reagent strip contained in the chromatography detection structure, and a second insulin-resistant antibody not marked is coated on a nitrocellulose membrane.

Description

Rapid detection structure for blood insulin level and application thereof

Technical Field

The invention relates to the technical field of medical instruments, in particular to a rapid detection structure for blood insulin level and application thereof.

Background

Insulin is a protein hormone secreted by the beta cells of the islets of langerhans in the pancreas, consisting of two peptide chains A and B, regulated by endogenous or exogenous substances such as glucose, lactose, ribose, arginine, glucagon, etc. Insulin is the most important protein hormone for promoting anabolism of human bodies, can promote glucose uptake and utilization of tissue cells for glycometabolism, promote glycogen synthesis, inhibit gluconeogenesis and reduce blood sugar; can promote fatty acid synthesis and fat storage for fat metabolism, and reduce fat decomposition; the protein can promote amino acid to enter cells, and promote each link of protein synthesis to increase protein synthesis.

The immunological detection technology is a blood insulin level detection method which is commonly used at present, and comprises an immunoturbidimetry method, a chemiluminescence method, an enzyme-linked immunosorbent assay, a chemiluminescence immunoassay method, a time-resolved immunofluorescence method, a quantum dot immunofluorescence chromatography method, a latex immunoturbidimetry method, a latex enhanced immunoturbidimetry method, a fluorescence immunochromatography method and the like. High sensitivity, rapidness, convenience, miniaturization, full quantification and automation are the development trend of the current clinical immunoassay technology. The point-of-care testing (POCT) is a branch of the current immunological testing technology which is the fastest developed, and the chromatographic immunoassay is the most commonly used testing method, wherein the products of colloidal gold chromatography, fluorescence chromatography and latex microsphere chromatography are used most widely, but the testing of blood insulin level is mainly based on immunoturbidimetry and chemiluminescence, and although the immunochromatography is also used, the integrated testing technology of sampling and testing result interpretation is not available, so that the testing is still limited to be operated and used by laboratories and professionals, and cannot be popularized and used to families and medical institutions lacking corresponding testing conditions. A large number of researches prove that insulin becomes protein hormone and important regulation indexes and markers which directly influence various metabolic related diseases including diabetes and bad health states, and is suitable for detection of various diseases and health states in and out of hospitals, so that the technology can be developed for detection of home self-detection and medical institutions lacking corresponding detection conditions, has a blood insulin level rapid detection technology which is simple and convenient to operate, rapid to use and low in cost, is beneficial to improvement of medical quality and efficiency, and has important clinical significance and application value.

Disclosure of Invention

Compared with the prior art, the invention has the advantages of convenient and quick detection operation, low cost and the like, and improves the detection quality.

In view of the above object, the present invention provides a structure for rapidly detecting blood insulin level, which is characterized in that:

1) the rapid detection structure is a combined set of a capillary siphon sampling structure, a detection liquid loading structure and a chromatography detection structure;

2) the capillary siphon sampling structure consists of a capillary siphon quantitative structure and a fixing structure for fixing the capillary siphon quantitative structure, wherein the capillary siphon quantitative structure and the fixing structure are fixedly connected in a non-dismounting manner, and one side of the fixing structure is of a non-closed structure;

3) the chromatography detection structure comprises a detection reagent strip, a detection shell positioned on the outer side of the detection reagent strip and a sample adding connection structure positioned at the starting end of the detection shell;

4) the detection reagent strip comprises a sample pad, a marker combination pad, a nitrocellulose membrane and a water absorption pad, wherein the sample pad is positioned at a corresponding position of the sample adding connection structure and forms liquid phase traffic connection with the sample adding connection structure, the marker combination pad is coated with a first insulin-resistant antibody marked by an indicator, and the nitrocellulose membrane is coated with a second insulin-resistant antibody which is not marked;

5) the capillary siphon quantitative structure is a tubular structure with two open ends, wherein one open end is connected with the sample pad, the quantitative volume is the volume of the collected blood sample, and 1-30 microliters is selected by taking the whole volume of the tubular structure as the quantitative volume of the sample;

6) the detection liquid loading structure is a liquid-phase-loading container structure provided with a liquid outlet for dripping liquid and is not directly and fixedly connected with the chromatography detection structure;

7) the detection liquid loading structure is loaded with a detection liquid phase, and the detection liquid phase is a water-soluble buffer salt solution.

The sample adding connection structure of the chromatography detection structure comprises a connection structure connected with the protective sleeve sample structure, and a sample adding window or a sample adding hole positioned on the chromatography detection structure.

The liquid outlet of the detection liquid loading structure for dripping liquid is dripper with different liquid outlet calibers, and the quantitative amount of the dripped liquid can be adjusted and controlled through the calibers.

In the above fast detection structure, the capillary siphon sampling structure and the chromatography detection structure are combined structures directly connected with each other, the capillary siphon quantitative structure is located at the proximal end of the chromatography detection structure and is fixedly connected, and the distal end opening of the capillary siphon quantitative structure is connected with the sample pad to form liquid phase traffic, the proximal end is in a free state and can extend the proximal end opening into the blood sample during sampling, and the chromatography detection structure is used as the fixing structure of the capillary siphon quantitative structure at the same time; the near-end is provided with dismantled and assembled protective sheath appearance structure, protective sheath appearance structure is provided with the application of sample hole, with the near-end opening of capillary siphon ration structure is located in the protective sheath, directly with the application of sample hole forms liquid phase traffic, the liquid outlet that detects liquid and load the structure is located the outside of protective sheath, directly with the application of sample hole forms dismantled and assembled liquid phase traffic.

In the above rapid detection structure, the capillary siphon sampling structure and the chromatography detection structure are combined structures that can move relative to each other to form a detachable connection, the capillary siphon quantitative structure and the fixing structure are directly and fixedly connected to form the capillary siphon sampling structure that can be sampled by hand, at this time, the fixing structure is used as a sampling handle during sampling, and the other end of the fixing structure is the capillary siphon quantitative structure in a free state and can extend the free end opening of the capillary siphon quantitative structure into a blood sample for sampling; the chromatography detection structure is characterized in that the near end of the chromatography detection structure is provided with the sample adding connecting structure, a sample adding opening is arranged at a position corresponding to the sample pad, liquid phase traffic is directly formed between the free end opening of the capillary siphon quantitative structure and the sample adding opening, and a liquid outlet of the detection liquid loading structure is directly formed between the liquid outlet of the detection liquid loading structure and the sample adding opening.

In the above-mentioned rapid detection structure, be provided with blood cell filtration structure on the detection reagent strip, including at least one of blood cell filtration membrane and erythrocyte antibody processing membrane pad.

In the rapid detection structure, the capillary siphon quantitative structure is a capillary dropwise adding quantitative tubular structure and consists of a handle and a dripper connected with the handle, and the cross section area of the opening end of the dripper is selected to be 0.10-3.0 square millimeters.

In the above rapid detection structure, the detection reagent strip is at least one of a colloidal gold immunoassay reagent strip, a fluorescence immunoassay reagent strip and a latex microsphere immunoassay reagent strip, and the indicator is at least one of a color microsphere and a fluorescent substance.

In the above-mentioned short-term test structure, the short-term test structure dispose with the indicator colour unanimous semi-quantitative colour comparison card, the colour comparison card is printed with the color comparison strip of 3 and above different colour depths.

In the above-mentioned rapid detection structure, the rapid detection structure is provided with a portable quantitative detector including at least one of a colorimetric quantitative analyzer and a fluorescent quantitative analyzer.

In the above rapid test structure, the rapid test structure comprises a lancet and an alcohol disinfectant tablet.

In the above rapid detection structure, the detection reagent strip further comprises a biotin/avidin reaction system, the marker binding pad is coated with a first insulin-resistant antibody labeled by the indicator and a second insulin-resistant antibody labeled by biotin, and the nitrocellulose membrane is coated with non-labeled avidin and analogues thereof. And then the insulin in the sample to be detected is simultaneously combined with the first insulin-resistant antibody marked by the indicator and the second insulin-resistant antibody marked by the biotin to form a first insulin-resistant antibody-insulin-biotin-marked second insulin-resistant antibody complex which is marked by the indicator, then the first insulin-resistant antibody-insulin-biotin-marked second insulin-resistant antibody complex flows through an avidin coated nitrocellulose membrane, is captured to form the first insulin-resistant antibody-insulin-biotin-marked second insulin-resistant antibody-avidin complex which is marked by the indicator, is gathered on a coating strip and displays visible color or fluorescence, and is used as a basis for quantification or semiquantification. Avidin and its analogs include avidin, streptavidin, and other substances that have specific binding to biotin.

In the above fast detecting structure, the operation of the fast detecting structure comprises the following steps:

1) taking out the chromatography detection structure, the capillary siphon sampling structure, the detection liquid loading structure, the lancet and the alcohol disinfection sheet, and exposing the sampling head at the opening end of the capillary siphon quantitative structure;

2) disinfecting the blood sampling part of the fingertip by using an alcohol disinfection tablet, and puncturing by using a lancet;

3) inserting the sampling head at the opening end of the capillary siphon quantitative structure into the blood sample, and naturally siphoning the blood sample and fully sucking the blood sample into the tubular structure of the capillary siphon quantitative structure;

4) transferring the blood sample in the tubular structure to a test reagent strip sample pad;

5) quantitatively adding a detection liquid to the sample pad, mixing the detection liquid with the blood sample, and allowing the mixture to flow forwards through the marker combination pad, the blood cell filtering structure, the nitrocellulose membrane and the water absorption pad;

6) and reading the detection result from the observation window to finish detection.

The rapid detection structure is applied to development of products for rapidly detecting blood insulin level.

In the above fast detecting structure, the operation of the fast detecting structure further includes the following steps:

3) inserting the sampling head at the open end of the capillary siphon quantitative structure into the blood sample, naturally siphoning the blood sample and fully sucking the tubular structure of the capillary siphon quantitative structure;

4) mixing the blood sample in the tubular structure with a detection solution, and quantitatively transferring the mixed solution to a detection reagent strip sample pad;

5) flowing through the label combining pad, the nitrocellulose membrane and the water absorption pad;

The detection reagent strip is a sample pad, a marker combination pad, a blood cell filtering structure, a nitrocellulose membrane and a water absorption pad which are sequentially adhered to a PVC negative sheet, the detection reagent strip is placed in a detection shell, and the detection shell is of a rigid structure or a flexible structure.

Due to the adoption of the technical scheme, the invention has the following advantages:

1. the invention relates to a rapid detection node for blood insulin level, which comprises a capillary siphon sampling structure, a combined set of a detection liquid loading structure and a chromatography detection structure, and a lancet and an alcohol disinfection sheet for sampling, and can complete the whole process from sample collection to detection at one time, thereby obviously improving the timeliness of clinical detection and the feasibility of popularization and promotion, and improving the clinical diagnosis and treatment effect.

2. The invention adopts a capillary siphon quantitative structure, controls the collection amount of the sample through the volume in the capillary siphon, collects trace amount, namely micro-upgrade amount, simultaneously adopts natural siphon in the collection process, does not need special quantitative equipment, ensures that the quantification is feasible and convenient to operate, and realizes convenient detection and quick operability of technical products.

3. The amount of the blood sample for detection is small, only 1-30 microliter, and is similar to that of the sample for detecting the peripheral blood sugar, so that the acceptance compliance of a detection technology product by a user is obviously improved.

4. The invention is provided with the colorimetric card for detecting the color intensity of the strip in semi-quantitative detection, can judge the semi-quantitative detection result by comparing the colorimetric cards, is suitable for clinical application scenes which do not need accurate quantification and only need to observe the change trend of the blood insulin level, and improves the clinical application value.

5. The invention is provided with the colorimetric card for detecting the color intensity of the strip in semi-quantitative detection, can judge the semi-quantitative detection result by comparing the colorimetric cards, is suitable for clinical application scenes which do not need accurate quantification and only need to observe the change trend of the blood insulin level, and improves the clinical application value.

6. The method has simple operation steps, is easy to realize household use or self-detection, is convenient to use, reduces the waste of raw materials, obviously improves the working efficiency, and can be applied to various fields of profession and amateur detection.

Drawings

FIG. 1 is a schematic view of the overall structure of the present invention;

FIG. 2 is a schematic longitudinal sectional view of a combined structure of a capillary siphon sampling structure and a chromatography detection structure according to the present invention;

FIG. 3 is a schematic view of the combination structure of the capillary siphon sampling structure and the chromatography detection structure in an integrated use state;

FIG. 4 is a schematic view of a capillary siphon quantitative sampling structure of the present invention;

FIG. 5 is a schematic view of a chromatographic assay according to the present invention;

FIG. 6 is a schematic structural view of a detachable combination of a capillary siphon sampling structure and a chromatography detection structure according to the present invention;

FIG. 7 is a schematic view of the loading structure of the detection solution of the present invention;

FIG. 8 is a schematic view of a detachable combination of a capillary siphon sampling structure and a detection liquid loading structure according to the present invention;

FIG. 9 is a schematic view of the test strip of the present invention;

FIG. 10 is a schematic view of a capillary drip quantitative sampling configuration of the present invention;

FIG. 11 is a schematic structural view of a detachable combination of a capillary dripping quantitative sampling structure and a chromatography detection structure in use according to the present invention;

FIG. 12 is a schematic structural view of a detachable combination of a detection solution loading structure and a chromatography detection structure according to the present invention;

FIG. 13 is a schematic diagram of a semi-quantitative colorimetric card according to the present invention.

The figures are labeled as follows:

a capillary siphon sampling structure 1; a detection liquid loading structure 2; a chromatographic detection structure 3; a capillary siphon quantitative tubular structure 4; a detection liquid loading container 5; detecting the liquid dropper 6; detecting liquid 7; a protective jacket structure 8; a sample addition window 9; detecting the connection structure 10; an observation window 11; a test reagent strip 12; a detection housing upper cover 13; a test housing base 14; a sampling handle 15; a capillary siphon-shaped fixed structure 16; a capillary siphon tubular structure traffic opening 17; a removable chromatographic detection structure base 18; a detachable chromatography detection structure upper cover 19; a detachable chromatographic detection structure sample adding hole 20; a detachable chromatographic detection structure observation window 21; a detection liquid loading container tube cap 22; a test strip sample pad 23; detection reagent strip label conjugate pad 24; a nitrocellulose membrane pad 25 of the detection reagent strip; a test reagent strip wicking pad 26; detecting a reagent strip PVC bottom sheet 27; a capillary drip quantitative tubular structure dripper 28; and a semi-quantitative color comparison card 29.

Detailed Description

To further illustrate the technical means and effects of the present invention adopted to achieve the predetermined purpose, the following embodiments are further described with reference to the accompanying drawings, but the present invention is not limited to the following description.

As shown in fig. 1, the whole structure of the invention comprises a capillary siphon sampling structure 1, a detection liquid loading structure 2 and a chromatography detection structure 3, wherein the capillary siphon sampling structure 1 comprises a capillary siphon quantitative tubular structure 4, and the quantitative sampling process is completed by the siphon constant volume process of the quantitative tubular structure 4; the detection liquid loading structure 2 comprises a detection liquid loading container 5, a detection liquid dropper 6 and detection liquid 7; chromatography detection structure 3 includes application of sample window 9, observation window 11, detection reagent strip 12, detects shell upper cover 13 and detects shell base 14, and detection reagent strip 12 places in the inner chamber that detects shell base 14 and detect shell upper cover 13 and the 9 application of sample of accessible application of sample window, and 11 observation testing results of observation window.

As shown in fig. 2, the capillary siphon sampling structure and the chromatography detection structure of the present invention are integrated into a combined structure, the capillary siphon quantitative tubular structure 4 is fixed by the proximal end of the chromatography detection structure 3 to form a fixed section 4A inside the chromatography detection structure 3 and a free section 4B outside the chromatography detection structure 3, the open end of the fixed section 4A is connected with the sample pad of the chromatography detection structure 3 to form liquid phase traffic, the free section 4B is in a free state and can extend the opening of the free end into the blood sample during sampling, and the chromatography detection structure 3 is used as the fixed structure of the capillary siphon quantitative structure at the same time; the near end is provided with a detachable protective sleeve sample structure 8, a sample adding hole 9 is arranged on the protective sleeve sample structure 8, and an opening of the near end (free end 4B) of the capillary siphon quantitative structure 4 is positioned in the protective sleeve sample structure 8 and directly forms liquid phase traffic with the sample adding hole 9.

As shown in fig. 3, the combined structure of the capillary siphon sampling structure and the chromatography detection structure in an integrated use state of the invention comprises a protective sleeve sample structure 8 inserted into a detection connection structure 10 to form an integrated structure of the capillary siphon sampling structure 1 and the chromatography detection structure 3, wherein an opening at a proximal end (a free end 4B) of a capillary siphon quantification structure 4 is positioned in a sample adding hole 9 to form liquid phase communication connection with the sample adding hole 9, and a detection result is observed through an observation window 11.

As shown in fig. 4, the capillary siphon quantitative sampling structure of the present invention includes a sampling handle 15, a capillary siphon-shaped fixing structure 16, a capillary siphon-shaped structure traffic opening 17, and a capillary siphon quantitative structure 4, wherein the capillary siphon quantitative structure 4 is a capillary siphon-shaped structure with two open ends, is fixed by the capillary siphon-shaped fixing structure 16 and is connected with the sampling handle 15, and the capillary siphon-shaped structure traffic opening 17 is disposed at the upper end of the sampling handle 15 to ensure that the non-sampling open end of the capillary siphon quantitative structure 4 is a non-closed structure. The capillary siphon quantitative structure 4 is fixedly connected with the fixing structure 16 for fixing the capillary siphon quantitative structure 4 in a non-dismounting way. One end opening of the capillary siphon quantitative structure 4 is connected with the sample pad of the detection reagent strip, and the other end opening extends into the blood sample to be detected for sampling, wherein the quantitative volume is the volume of the collected blood sample, and 1-30 microliters is selected by taking the whole volume of the capillary tube structure as the quantitative volume of the sample.

As shown in fig. 5 and fig. 6, the chromatography detection structure of the present invention and the structure forming a detachable combination using state with the capillary siphon sampling structure, wherein the chromatography detection structure includes a detachable chromatography detection structure base 18 and a detachable chromatography detection structure upper cover 19, a detachable chromatography detection structure sample adding hole 20 and a detachable chromatography detection structure observation window 21 arranged on the detachable chromatography detection structure upper cover 19, and a detection reagent strip 12 placed on the detachable chromatography detection structure base 18 at a position corresponding to the sample adding hole 20 and the observation window 21; the capillary siphon sampling structure comprises a sampling handle 15, a capillary siphon-shaped fixing structure 16, a capillary siphon quantitative tubular structure 4 fixedly connected with the fixing structure 16 and a capillary siphon tubular structure traffic opening 17 positioned at the upper end of the sampling handle 15; the structure of the detachable combined use state is the detachable liquid phase traffic connection between the capillary siphon quantitative tubular structure 4 and the detachable chromatography detection structure sample adding hole 20. When the portable blood sampling device is used specifically, the chromatographic detection structure is placed on the plane, the sampling handle 15 is held by hands, the traffic open hole 17 is kept in an open state, the free end opening of the capillary siphon quantitative tubular structure 4 is inserted into a blood sample, the blood sample is naturally siphoned to fill the quantitative tubular structure 4, the quantitative tubular structure 4 is moved to the detachable chromatographic detection structure sample adding hole 20, and all blood samples are released to the detection reagent strips 12 in the sample adding hole.

As shown in fig. 7 and 12, the structure of the present invention for loading detection solution and the structure of the detachable combination using state with the chromatography detection structure, wherein the detection solution loading structure 2 comprises a detection solution loading container 5, a detection solution dropping head 6 at the top end, detection solution 7 loaded in the detection solution loading container 5 and a detection solution loading container tube cover 22; the detachable combination using state is structured by the detachable liquid phase traffic connection between the detection liquid dropper 6 and the detachable chromatography detection structure sample adding hole 20. When the detection device is used specifically, the chromatography detection structure is placed on a plane, the detection liquid loading container 5 is held by hands, the direction of the detection liquid dropping head 6 is kept downward to the sample adding hole 20 of the detachable chromatography detection structure, and the detection liquid 7 is dropped to the detection reagent strip 12 in the sample adding hole.

As shown in fig. 7 and 8, the detachable combination structure of the capillary siphon sampling structure and the detection liquid loading structure of the present invention is a detachable liquid phase communication connection between the capillary siphon quantitative tubular structure 4 and the detection liquid loading container 5. When the blood sampling device is used specifically, the detection liquid loading container 5 is placed on a plane, the sampling handle 15 is held by a hand, the traffic open hole 17 is kept in an open state, the free end opening of the capillary siphon quantitative tubular structure 4 is inserted into a blood sample, the blood sample is naturally siphoned to fully suck the quantitative tubular structure 4, the quantitative tubular structure 4 is moved to the detection liquid 7 of the detection liquid loading container 5, and all blood samples are released into the detection liquid 7.

As shown in fig. 1, fig. 2, fig. 5 and fig. 9, the detection reagent strip structure of the present invention comprises a detection reagent strip sample pad 23, a detection reagent strip marker binding pad 24, a detection reagent strip nitrocellulose membrane pad 25 and a detection reagent strip water absorption pad 26, which are sequentially attached to a detection reagent strip PVC negative 27 from near to far, the position sequentially corresponding to the chromatographic detection structure is the corresponding position of the detection reagent strip sample pad 23 on the sample application window 9 or the sample application hole 20 of the detachable chromatographic detection structure, and the detection reagent strip nitrocellulose membrane pad 25 is the corresponding position of the observation window 11 or the observation window 21 of the detachable chromatographic detection structure; the detection reagent strip marker conjugate pad 24 is coated with a colloidal gold particle, a colored latex particle or a fluorescent microsphere and an indicator labeled conjugate of fluorescein, which are labeled by a first anti-insulin antibody; the nitrocellulose membrane pad 25 of the detection reagent strip is coated with a non-labeled second anti-insulin antibody.

As shown in fig. 10 and 11, the capillary dripping quantitative sampling structure of the present invention and the structure of the chromatography detection structure in a detachable combination use state comprise a sampling handle 15, a fixing structure 16 and a capillary dripping quantitative tubular structure dropper 28, wherein the capillary dripping quantitative tubular structure dropper 28 is a capillary tubular structure with an open end, is fixed by the fixing structure 16 and is connected with the sampling handle 15, and the cross-sectional area of the end of the dropper 28 is selected from 0.10 to 3.0 square millimeters. When the chromatography detection structure is used specifically, the chromatography detection structure plane is placed, the sampling handle 15 is held by hand, the capillary dropwise adding quantitative tubular structure dripper 28 is inserted into a blood sample, a certain amount of blood sample to be detected is absorbed, the capillary dropwise adding quantitative tubular structure dripper 28 is connected to the detachable chromatography detection structure sample adding hole 20, and a certain drop number of blood sample is dropwise added to the detection reagent strip 12 in the sample adding hole.

As shown in fig. 13, the semi-quantitative color chart 29 of the present invention is a color chart printed with 10 color lines of the same type as the indicator with different gradient color depths.

Thus, in practical operation, when the rapid detection structure is a colloidal gold immunoassay structure, the detection reagent strip 11 is prepared by a colloidal gold method, and a sample pad, a colloidal gold binding pad coated with a colloidal gold-labeled first anti-insulin antibody, a blood cell filtering structure, a nitrocellulose membrane pad coated with a non-labeled second anti-insulin antibody and a water absorption pad are sequentially adhered on the PVC bottom sheet; when the rapid detection structure is a latex microsphere immunoassay structure, the detection reagent strip 11 is prepared by a latex microsphere immunoassay method, and a sample pad, a latex microsphere combined pad coated with a latex microsphere marker, a blood cell filtering structure, a nitrocellulose membrane pad coated with a non-labeled capture reagent and a water absorption pad are sequentially adhered on a PVC (polyvinyl chloride) negative film; when the rapid detection structure is a fluorescence immunoassay structure, the detection reagent strip 11 is prepared by taking fluorescent microspheres or fluorescein as an indicator through a fluorescence immunoassay method, and a sample pad, a latex microsphere combination pad coated with a latex microsphere marker, a blood cell filtering structure, a nitrocellulose membrane pad coated with a non-labeled capture reagent and a water absorption pad are sequentially adhered on a PVC (polyvinyl chloride) bottom sheet.

The semi-quantitative detection specific operation comprises the following steps: 1) taking out the chromatography detection structure 3, the capillary siphon sampling structure 1, the detection liquid loading structure 2, the semi-quantitative colorimetric card 29, the lancet and the alcohol disinfection sheet, and exposing the sampling head at the opening end of the capillary siphon quantitative structure; 2) disinfecting the blood sampling part of the fingertip by taking an alcohol disinfection tablet, and puncturing by using a lancet; 3) inserting the sampling head 4B at the opening end of the capillary siphon quantitative structure into the blood sample, naturally siphoning the blood sample and fully sucking the capillary siphon quantitative structure; 4) transferring the blood sample in the tubular structure to a test reagent strip sample pad; 5) quantitatively adding a detection liquid phase to the sample pad, mixing with the blood sample, and allowing the mixture to flow forwards through the marker combination pad, the blood cell filtering structure, the nitrocellulose membrane and the water absorption pad; 6) and reading the detection result from the observation window, comparing the detection result with the semi-quantitative colorimetric card 29, reading the semi-quantitative range value, and finishing the detection.

The specific operation of quantitative detection comprises the following steps: 1) taking out the chromatography detection structure 3, the capillary siphon sampling structure 1, the detection liquid loading structure 2, the quantitative detector, the lancet and the alcohol disinfection sheet, and exposing the sampling head at the opening end of the capillary siphon quantitative structure; 2) disinfecting the blood sampling part of the fingertip by using an alcohol disinfection tablet, and puncturing by using a lancet; 3) inserting the sampling head 4B at the opening end of the capillary siphon quantitative structure into the blood sample, naturally siphoning the blood sample and fully sucking the capillary siphon quantitative structure; 4) transferring the blood sample in the tubular structure to a test reagent strip sample pad; 5) quantitatively adding a detection liquid phase to the sample pad, mixing with the blood sample, and allowing the mixture to flow forwards through the marker combination pad, the blood cell filtering structure, the nitrocellulose membrane and the water absorption pad; 6) and (3) placing the chromatographic detection structure 3 in a quantitative detector, reading the detection result, and quantitatively calculating the measured value to finish the detection.

Experimental study of the invention: the following experiment is illustrative of the detection method of the present invention and its effects, but is not intended to limit the present invention. The experimental methods used in the following experiments are all conventional methods unless otherwise specified. The materials, reagents and the like used are commercially available unless otherwise specified.

Experiment one: immune colloidal gold method blood insulin rapid detection experiment:

firstly, preparing a detection reagent strip:

the detection reagent strip is prepared by adopting a conventional immune colloidal gold detection technology and a double-antibody sandwich method, the detection reagent kit is prepared by adopting the rapid detection structure to carry out an insulin detection experiment, wherein a colloidal gold mark indication antibody of a detection line T of the detection reagent strip is an insulin-resistant monoclonal antibody of 10ug/ml, and colloidal gold particles with the particle size of 50nm are coated on a glass cellulose membrane colloidal gold combination pad; the capture antibody of the detection line T of the detection reagent strip is 1.0mg/ml of paired anti-insulin monoclonal antibodies, and the capture antibody is coated on the nitrocellulose membrane pad; the capture antibody of the quality control line C of the detection reagent strip is a goat anti-mouse IgG polyclonal antibody of 1.0mg/ml, and the capture antibody is coated on a nitrocellulose membrane pad and used for capturing the colloidal gold labeled anti-insulin monoclonal antibody which is not captured specifically. And respectively sticking a water absorption pad and a colloidal gold mark combination pad at two ends of the nitrocellulose membrane pad printing membrane, and sticking a sample pad at one side of the colloidal gold mark combination pad. And placing the adhered detection sheet on a strip cutting machine, and cutting into detection reagent strips with the thickness of 3.5 mm.

Secondly, preparing a rapid detection structure:

the fast detection structure provided by the invention is designed by Solidworks, the sample structures of the upper cover, the base and the protective sleeve of the shell are detected, a sample is printed in 3D, a commercially available capillary tube is cut into 10mm and about 10ul, a storage tube used in a small experiment is used as a diluent loading container, and the prepared fast detection structure sample is used for experimental detection. Printing red color comparison cards with different color depths.

Third, experimental method and result:

during the experiment, the prepared detection reagent strip and the rapid detection structure are assembled into an integrated detection structure, the assembled detection structure is placed into an aluminum foil sealing bag with a drying agent, the aluminum foil sealing bag is sealed on a sealing machine, and a label is added. And printing 10 purple color comparison cards with different color depths. 1, 5, 10, 15, 20, 25 and 30uIU/ml of ChangxuiLin recombinant insulin glargine injection (100 IU/ml) is prepared by 10mM phosphate buffer salt solution. Taking 100ul of insulin detemir solution, directly dripping insulin detemir solution with different concentrations to a sample adding window of a chromatography detection structure, standing for 20 minutes, checking the color depth of a detection line through an observation window, carrying out colorimetry with a semi-quantitative colorimetric card, judging the concentration range corresponding to the colorimetric result, wherein the purple red line No. 1 of the colorimetric card corresponds to 5uIU/ml, No. 2 is 10uIU/ml, No. 3 is 15uIU/ml, No. 4 is 20uIU/ml, No. 5 is 25uIU/ml, and No. 6 is 30 uIU/ml.

10ul of fasting and 2-hour postprandial peripheral blood of 10 healthy donors are respectively collected by using siphon sampling tubes of prepared rapid detection structure samples, the fasting and 2-hour postprandial peripheral blood are respectively and directly loaded to a sample loading window of a chromatography detection structure, the mixture is stood for 20 minutes, the color depth of a detection line is checked through an observation window, the mixture is compared with a semi-quantitative colorimetric card, the concentration range of a colorimetric result is judged, the fasting result is about 5-15uU/ml, the 2-hour postprandial color is about 15-25uU/ml, and the detection range of normal people is met.

Experiment two: the fast detection experiment of insulin by latex microsphere immunochromatography comprises the following steps:

firstly, preparing a detection reagent strip:

the detection reagent strip is prepared by a double-antibody sandwich method of a conventional latex microsphere immunochromatography technology, the detachable chromatography detection structure detection kit is prepared by the rapid detection structure to carry out an insulin detection experiment, latex microspheres all adopt biological red microspheres with the particle size of 300nm, and an anti-insulin monoclonal antibody with the antibody of 50ug/ml is marked by the latex microspheres of a detection line T of the detection reagent strip and is coated on a glass cellulose membrane bonding pad; the capture antibody of the detection line T of the detection reagent strip is a paired anti-insulin monoclonal antibody of 1.0mg/ml, and is coated on the nitrocellulose membrane pad; the capture antibody of the quality control line C of the detection reagent strip is a goat anti-mouse IgG polyclonal antibody of 1.0mg/ml, and the capture antibody is coated on the nitrocellulose membrane pad and used for capturing latex microsphere labeled anti-insulin monoclonal antibody which is not captured specifically. And respectively sticking a water absorption pad and a latex microsphere mark combination pad at two ends of the nitrocellulose membrane pad printing membrane, and sticking a sample pad at one side of the latex microsphere mark combination pad. And placing the adhered detection sheet on a strip cutting machine, and cutting into detection reagent strips with the thickness of 3.5 mm.

Secondly, preparing a rapid detection structure:

using commercially available capillaries and droppers, the commercially available capillaries were cut to 10mm, about 10ul, and about half of the length was inserted into the dropper from the dropper head, leaving the other half outside the dropper as the free end. The detachable chromatography detection structure adopts a commercially available chromatography detection buckle. The detection liquid loading structure adopts a storage tube used in a small-scale experiment. Printing red color comparison cards with different color depths.

Third, experimental method and result:

during the experiment, the prepared detection reagent strip and the rapid detection structure are assembled into an integrated detection structure, the assembled detection structure is placed into an aluminum foil sealing bag with a drying agent, the aluminum foil sealing bag is sealed on a sealing machine, and a label is added. And printing 10 purple color comparison cards with different color depths. 1, 5, 10, 15, 20, 25, 30uIU/ml of ChanxiuLin recombinant insulin glargine injection (100 IU/ml) is prepared by 10mM phosphate buffered saline solution. Taking 100ul of insulin detemir solution, directly dripping insulin detemir solution with different concentrations to a sample adding window of a chromatography detection structure, standing for 20 minutes, checking the color depth of a detection line through an observation window, carrying out colorimetry with a semi-quantitative colorimetric card, judging the concentration range corresponding to the colorimetric result, wherein the purple red line No. 1 of the colorimetric card corresponds to 5uIU/ml, No. 2 is 10uIU/ml, No. 3 is 15uIU/ml, No. 4 is 20uIU/ml, No. 5 is 25uIU/ml, and No. 6 is 30 uIU/ml.

10ul of fasting and 2-hour postprandial peripheral blood of 10 healthy donors are respectively collected by using siphon sampling tubes of prepared samples with rapid detection structures, the samples are directly and respectively loaded to a sample loading window of a chromatography detection structure, the samples are stood for 20 minutes, the color depth of a detection line is checked through an observation window, the samples are compared with a semi-quantitative colorimetric card, the concentration range of a colorimetric result is judged, the fasting result is 5-15uIU/ml, the 2-hour postprandial result is about 15-25uIU/ml, and the detection range of normal people is met.

Experiment three: an immunofluorescence method insulin rapid detection experiment:

firstly, preparing a detection reagent strip:

the detection reagent strip is prepared by adopting a conventional immunofluorescence detection technology double-antibody sandwich method and a biotin/avidin reaction system, and the detachable chromatography detection structure detection kit prepared by adopting the rapid detection structure is used for carrying out an insulin detection experiment, wherein a fluorescent microsphere marking indication antibody of the detection reagent strip adopts a first insulin-resistant monoclonal antibody of 20ug/ml, a biotinylation marking antibody adopts a second insulin-resistant monoclonal antibody of 20ug/ml, the fluorescent microsphere adopts a particle size of 300nm, and the two marking antibodies are both coated on a glass cellulose membrane marker combination pad; the capture antibody of the detection reagent strip is 1.0mg/ml of recombinant streptavidin, and is coated on a nitrocellulose membrane; the capture antibody of the quality control line C of the detection reagent strip is a goat anti-mouse IgG polyclonal antibody of 1.0mg/ml, and the capture antibody is coated on a nitrocellulose membrane and used for capturing the fluorescent microsphere labeled anti-insulin monoclonal antibody which is not captured specifically. And respectively sticking a water absorption paper membrane pad and a fluorescent microsphere mark combined membrane pad at two ends of the nitrocellulose membrane printing membrane, and sticking a sample pad at one side of the combined membrane pad. And placing the adhered detection sheet on a strip cutting machine, and cutting into 3.5mm test strips.

Secondly, preparing a rapid detection structure:

the same as experiment two.

Thirdly, a fluorescence detector: a commercially available blue Bo AFS-100 fluorescence detector was used.

Fourthly, experimental method and result:

during the experiment, the prepared detection reagent strip and the rapid detection structure are assembled into a detection kit, the assembled detection kit is put into an aluminum amber sealing bag with a drying agent, the sealing is carried out on a sealing machine, and a label is added. Preparing 1, 5, 10, 15, 20, 25 and 30uIU/ml ChanxiuLin recombinant insulin glargine injection (100 IU/ml) by using 10mM phosphate buffer solution, and establishing a standard curve of insulin detemir concentration and fluorescence reaction by using the prepared detection kit and a fluorescence quantitative analyzer. 10ul of terminal blood of 10 healthy donors after fasting and 2 hours after meal are respectively collected by the prepared capillary siphon sampling tube, the terminal blood is directly and respectively loaded to a sample loading window of the prepared detection kit, the terminal blood is kept still for 20 minutes, the terminal blood is placed in a fluorescence quantitative analyzer to detect the fluorescence value, the insulin concentration of the sample blood is calculated by a standard curve, the result is 8.42 +/-2.70 uIU/ml for fasting and 17.27 +/-4.31 uIU/ml for 2 hours after meal, and the result accords with the detection range of normal people.

Claims

1. A quick detection structure of blood insulin level which characterized in that:

4) the detection reagent strip comprises a sample pad, a marker combination pad, a nitrocellulose membrane and a water absorption pad, wherein the sample pad is positioned at a corresponding position of the sample adding connection structure and forms liquid phase traffic connection with the sample adding connection structure, a first insulin-resistant antibody marked by an indicator is coated on the marker combination pad, and a second insulin-resistant antibody which is not marked is coated on the nitrocellulose membrane;

2. The rapid detection structure according to claim 1, wherein the capillary siphon sampling structure and the chromatographic detection structure are combined structures directly connected with each other, the capillary siphon quantitative structure is located at the proximal end of the chromatographic detection structure and fixedly connected with the chromatographic detection structure, the distal end opening of the capillary siphon quantitative structure is connected with the sample pad and forms liquid phase communication, the proximal end is in a free state and can extend the proximal end opening into the blood sample during sampling, and the chromatographic detection structure is used as the fixed structure of the capillary siphon quantitative structure; the near-end is provided with dismantled and assembled protective sheath appearance structure, protective sheath appearance structure is provided with the application of sample hole, with the near-end opening of capillary siphon ration structure is located in the protective sheath, directly with the application of sample hole forms liquid phase traffic, the liquid outlet that detects liquid and load the structure is located the outside of protective sheath, directly with the application of sample hole forms dismantled and assembled liquid phase traffic.

3. The rapid detection structure according to claim 1, wherein the capillary siphon sampling structure and the chromatographic detection structure are combined structures that can move relative to each other to form a detachable connection, the capillary siphon quantitative structure and the fixing structure are directly and fixedly connected to form the capillary siphon sampling structure that can be sampled by hand, at this time, the fixing structure is used as a sampling handle during sampling, and the other end of the fixing structure is in a free state and can extend a free end opening of the capillary siphon quantitative structure into a blood sample for sampling; the chromatography detection structure is characterized in that the near end of the chromatography detection structure is provided with the sample adding connecting structure, a sample adding opening is arranged at a position corresponding to the sample pad, liquid phase traffic is directly formed between the free end opening of the capillary siphon quantitative structure and the sample adding opening, and a liquid outlet of the detection liquid loading structure is directly formed between the liquid outlet of the detection liquid loading structure and the sample adding opening.

4. The rapid test structure according to claim 1, wherein the test reagent strip is provided with a hemocyte filtration structure comprising at least one of a hemocyte filtration membrane and a erythrocyte antibody treatment membrane pad.

5. The rapid detection structure of claim 1, wherein the capillary siphon quantitative structure is a capillary drip quantitative tubular structure, and comprises a handle and a dripper connected with the handle, and the cross-sectional area of the opening end of the dripper is selected to be 0.10-3.0 square millimeters.

6. The rapid detection structure of claim 1, wherein the detection reagent strip is at least one of a colloidal gold immunoassay reagent strip, a fluorescence immunoassay reagent strip, and a latex microsphere immunoassay reagent strip, and the indicator is at least one of a colored microsphere and a fluorescent substance.

7. The rapid detection structure of claim 1, wherein the rapid detection structure is configured with a semi-quantitative color comparison card having a color consistent with the indicator, and the color comparison card has 3 or more color comparison strips with different color depths printed thereon.

8. The rapid detection architecture of claim 1, wherein the rapid detection architecture is configured with a portable quantitative detector comprising at least one of a colorimetric quantitative analyzer and a fluorometric quantitative analyzer.

9. The rapid test structure of claim 1, wherein the rapid test structure comprises a lancet and an alcohol disinfectant tablet.

10. The rapid detection structure of claim 1, wherein the detection reagent strip is prepared by further comprising a biotin/avidin reaction system, the label binding pad is coated with a first anti-insulin antibody labeled with the indicator and a second anti-insulin antibody labeled with biotin, and the nitrocellulose membrane is coated with non-labeled avidin and its analogs.

11. The rapid detection architecture of claim 1, wherein the operational use of the rapid detection architecture comprises the steps of:

5) quantitatively adding a detection liquid phase to the sample pad, mixing with the blood sample, and allowing the mixture to flow forwards through the marker combination pad, the blood cell filtering structure, the nitrocellulose membrane and the water absorption pad;

12. The use of the rapid test structure of claim 1 in the development of a rapid test product for blood insulin levels.