Chemical-electrochemical luminescence common detection equipment and method

Abstract

The invention relates to the technical field of luminescence detection, and discloses chemical-electrochemical luminescence common detection equipment and a chemical-electrochemical luminescence common detection method. The chemical-electrochemical luminescence common detection equipment comprises a reaction cup, a plurality of reagent containers, a detection needle, a sample injection control module, a measuring cell, an electrode assembly, a magnetic adsorption piece and at least one set of optical detection module. The chemical-electrochemical luminescence common detection equipment is configured as follows: the detection needle sucks a sample mixture into the flow channel; after the conjugate is adsorbed by the magnetic adsorption part, the detection needle sucks the luminous reagent into the flow channel, and when the sucked luminous reagent is a chemiluminescence co-reaction reagent, chemiluminescence is realized; and when the absorbed luminescent reagent is an electrochemical luminescence co-reaction reagent, the electrode assembly is electrified to realize electrochemical luminescence. The invention further provides a chemiluminescence detection method which can be implemented through the device. According to the method, chemiluminescence detection can be carried out in the electrochemical luminescence detection equipment, and the accuracy is high.

Description

Technical Field

[0001] The present invention relates to the technical field of luminescence detection, and in particular to a chemical-electrochemical luminescence co-detection device and method. Background Art

[0002] Electrochemiluminescence (ECL) uses electrochemiluminescence, which requires voltage to be applied to stimulate chemiluminescence. It has the advantages of high sensitivity, low background, long luminescence time of luminescent material regeneration, and precise control of luminescence time. Electrochemiluminescence containers are usually carried out in closed flow channels to ensure consistent electrochemiluminescence conditions. The fixed luminescence container is called a measurement cell.

[0003] The chemiluminescence immunoassay analyzer is usually composed of one or more of a sample addition module, a reaction module, an optical detection module (photomultiplier tube), a data processing module, an incubation temperature control module, and a cleaning and separation module. The principle is generally to convert the light signal emitted by the chemiluminescence reaction into a digital signal, and the concentration value is calculated by the data processing system. Used in conjunction with adapter reagents, it is used for qualitative or quantitative analysis of the analyte in human samples. At present, the chemiluminescence detection method cannot be realized in the electrochemiluminescence detection device, which is generally a single test item each time; in addition, when there is an integrated machine with two chemiluminescence methodologies, or compatible with any chemiluminescence system, there is no suitable detection method to realize the electrochemical and chemiluminescence integrated machine. The integrated machine generally emits light in the reaction cup.

[0004] In view of this, this application is hereby filed. Summary of the invention

[0005] The objects of the present invention include, for example, providing a chemical-electrochemical luminescence co-detection device and method, aiming to improve at least one of the problems mentioned in the background art.

[0006] The embodiments of the present invention can be implemented as follows:

[0007] In a first aspect, the present invention provides a chemical-electrochemical luminescence co-detection device, comprising:

[0008] A reaction cup, a plurality of reagent containers, a detection needle, an injection control module, a measuring cell, an electrode assembly, a magnetic adsorption component and at least one set of optical detection modules;

[0009] The reaction cup is used to hold a sample mixture, the sample mixture includes a conjugate, the conjugate is mainly obtained by specifically binding at least two analytes in the sample to be tested to at least two binding molecules in a one-to-one correspondence and adsorbing them onto magnetic beads, at least one of the at least two binding molecules is a molecule labeled with a chemiluminescent marker, and at least one of the at least two binding molecules is a molecule labeled with an electrochemiluminescent marker;

[0010] At least a portion of the multiple reagent holding containers are used to hold multiple luminescent reagents respectively, and the multiple luminescent reagents include at least one chemiluminescent co-reactant and at least one electrochemiluminescent co-reactant;

[0011] The detection needle is used to draw the sample mixture from the reaction cup into the flow channel of the measuring cell, and is used to draw the corresponding luminescent reagents from multiple reagent holding containers into the flow channel of the measuring cell;

[0012] The sample injection control module is used to control the detection needle to absorb into the reaction cup and multiple reagent holding containers;

[0013] The measuring cell is used to provide a reaction space for realizing chemiluminescence and electrochemiluminescence;

[0014] The electrode assembly is arranged in conjunction with the measuring cell, and the electrode assembly is used to excite the electrochemical luminescent marker to emit light after being energized;

[0015] The magnetic adsorption element is arranged close to the measuring cell and is used to realize magnetic adsorption of magnetic beads in the flow channel entering the measuring cell;

[0016] At least one set of optical detection modules is used to detect and collect the luminous intensity of the luminescent marker;

[0017] The chemiluminescence co-detection device is configured as follows:

[0018] The sampling control module controls the detection needle to absorb the sample mixture into the flow channel of the measuring cell;

[0019] After the object to be bound is adsorbed by the magnetic adsorbent, the injection control module controls the detection needle to absorb the luminescent reagent into the flow channel of the measuring cell; when the absorbed luminescent reagent is a chemiluminescent co-reaction reagent, the chemiluminescent co-reaction reagent combines with the corresponding luminescent marker to achieve chemiluminescence, and the luminescence intensity is detected and collected by at least one set of optical detection modules; when the absorbed luminescent reagent is an electrochemiluminescent co-reaction reagent, the electrode assembly is energized, the electrochemiluminescent co-reaction reagent combines with the corresponding luminescent marker to achieve electrochemiluminescence, and the luminescence intensity is detected and collected by at least one set of optical detection modules.

[0020] In an optional embodiment, the chemi-electrochemiluminescence co-detection device further includes a control system;

[0021] The control system is in communication with the injection control module and the electrode assembly. The control system has a built-in timing control program. The timing control program is configured to instruct the injection control module to control the detection needle to absorb the corresponding reagent from different reagent containers at different time points.

[0022] When the current time node is the time node for absorbing the electrochemiluminescent co-reaction reagent, the control system sends a command to the electrode assembly to power it on. After the electrochemiluminescent reaction is completed, the control system controls the electrode assembly to power off.

[0023] In an optional embodiment, the timing control program is configured to: control the sampling control module to make the detection needle first sample the chemiluminescent co-reaction reagent to achieve chemiluminescence, and then sample the electrochemiluminescent co-reaction reagent to achieve electrochemiluminescence;

[0024] Alternatively, the timing control program is configured to: manipulate the injection control module to cause the detection needle to first sample the electrochemiluminescent co-reaction reagent to achieve electrochemiluminescence, and then sample the chemiluminescent co-reaction reagent to achieve chemiluminescence.

[0025] In an optional embodiment, some of the multiple reagent containers are used to hold neutral cleaning liquid; the timing control program is configured as follows:

[0026] Controlling the sampling control module to make the detection needle sequentially perform sampling of the sample mixture, absorption of the neutral cleaning solution, sampling of the chemiluminescent co-reaction reagent, and sampling of the electrochemiluminescent co-reaction reagent;

[0027] Alternatively, the sampling control module is manipulated to enable the detection needle to sequentially perform sampling of the sample mixture, absorption of the neutral cleaning solution, sampling of the electrochemiluminescent co-reaction reagent, and sampling of the chemiluminescent co-reaction reagent.

[0028] In an optional embodiment, the detection needle is also used to draw air into the flow channel of the measuring cell to form bubbles in the flow channel; the timing control program is configured as follows:

[0029] Manipulate the sampling control module to make the detection needle sample the sample mixture, absorb the neutral cleaning solution, absorb air, sample the chemiluminescent co-reaction reagent, and sample the electrochemiluminescent co-reaction reagent in sequence;

[0030] Alternatively, the sampling control module is manipulated to make the detection needle sample the sample mixture, absorb the neutral cleaning solution, absorb air, sample the electrochemiluminescent co-reaction reagent, and sample the chemiluminescent co-reaction reagent in sequence.

[0031] In an optional embodiment, a receiving groove is provided at the bottom of the outer wall of the measuring cell, and the receiving groove is used to receive at least part of the magnetic adsorption member; the magnetic adsorption member includes the following feature (1) or feature (2):

[0032] (1) The magnetic adsorbent is a permanent magnet, and the chemical-electrochemical luminescence co-detection device further includes a movement control module, which controls the magnetic adsorbent to move into the receiving groove to adsorb the magnetic beads at the beginning of the luminescence detection, and controls the magnetic adsorbent to move out of the receiving groove after the luminescence detection is completed to release the magnetic adsorbent from adsorbing the magnetic beads;

[0033] (2) The magnetic adsorption member is an electromagnet, and the electromagnet is configured to be powered on when the luminescence detection starts and powered off after the luminescence detection ends.

[0034] In an optional embodiment, the number of optical detection modules is 1 set, and the luminescent wavelengths of the luminescent markers used for detection can all be detected and collected by the optical detection module;

[0035] Alternatively, there are multiple sets of optical detection modules, and multiple luminescent markers are used for detection. The multiple luminescent markers have different luminescent wavelengths, and the luminous intensity of light emitted by the luminescent markers with different luminescent wavelengths is detected and collected by the corresponding optical detection modules respectively.

[0036] In an optional embodiment, a transparent window is provided on the upper wall of the measuring cell, at least one set of optical detection modules is provided above the transparent window, and a filter of a corresponding wavelength is provided between each set of optical detection modules and the transparent window;

[0037] Optionally, the filter is a narrow-band filter lens.

[0038] In a second aspect, the present invention provides a chemi-electrochemiluminescence detection method, comprising the following steps:

[0039] A sample mixture is introduced into the flow channel of the measuring cell, wherein the sample mixture includes a conjugate, which is mainly obtained by fully reacting the sample to be tested, at least one first binding molecule, at least one second binding molecule and magnetic beads, and the conjugate is adsorbed on the bottom of the flow channel when passing near the magnetic adsorption member arranged outside the flow channel;

[0040] A plurality of luminescent reagents are introduced into the flow channel of the measuring cell, each luminescent reagent is introduced at a different time point, and the plurality of luminescent reagents include at least one chemiluminescent co-reaction reagent and at least one electrochemiluminescent co-reaction reagent;

[0041] Each first binding molecule is a molecule labeled with a chemiluminescent marker. When there are multiple first binding molecules in the sample mixture, the multiple first binding molecules are molecules labeled with different chemiluminescent markers, and each first binding molecule is a substance that can specifically bind to a corresponding analyte in the sample to be tested and adsorb to the magnetic beads;

[0042] Each second binding molecule is a molecule labeled with an electrochemiluminescent marker. When the number of second binding molecules in the sample mixture is multiple, the multiple second binding molecules are molecules labeled with different electrochemiluminescent markers, and each second binding molecule is a substance that can specifically bind to a corresponding analyte in the sample to be tested and adsorbed to the magnetic beads;

[0043] At least one chemiluminescent co-reaction reagent is combined with the luminescent marker on at least one first binding molecule in a one-to-one correspondence to realize chemiluminescence, and the corresponding luminescence intensity is detected and collected by the corresponding optical detection module;

[0044] When the electrochemiluminescent co-reaction reagent is passed into the flow channel, the electrode assembly in the measuring cell is energized, and at least one electrochemiluminescent co-reaction reagent is combined with the luminescent marker on at least one second binding molecule in a one-to-one correspondence to realize electrochemiluminescence, and the corresponding luminescence intensity is detected and collected by the corresponding optical detection module.

[0045] In an optional embodiment, before introducing different luminescent reagents into the flow channel of the measuring cell, the method further includes: introducing a neutral cleaning solution into the flow channel of the measuring cell.

[0046] In an optional embodiment, when the conjugate is adsorbed to the bottom of the flow channel by the magnetic adsorbent, a neutral cleaning solution is introduced into the flow channel to wash away free substances in the sample mixture that are not bound to the magnetic beads, and then a luminescent reaction reagent is introduced.

[0047] In an optional embodiment, after the free substances not bound to the magnetic beads are washed away, before the luminescent reaction reagent is introduced, the process further includes introducing bubbles into the flow channel to isolate the neutral cleaning solution and the luminescent reaction reagent.

[0048] In an optional embodiment, the flow rate of the sample mixture in the flow channel is 10 μL / S to 30 μL / S, the flow rate of the chemiluminescent co-reaction reagent in the flow channel is 100 μL / S to 500 μL / S, and the flow rate of the electrochemiluminescent co-reaction reagent in the flow channel is 100 μL / S to 200 μL / S.

[0049] In an optional embodiment, it also includes at least one of the following features (1) to (6);

[0050] (1) After all luminescence detections are completed, an alkaline cleaning solution and an electrochemical luminescence co-reaction reagent are sequentially introduced into the flow channel, wherein the alkaline cleaning solution is a solution containing a detergent and a strong base, wherein the mass percentage of the detergent is 0.05% to 1%, the detergent is selected from at least one of Tween, Triton and polycatalpol, the strong base is selected from at least one of KOH and NaOH, and the pH of the alkaline cleaning solution is 12 to 14;

[0051] (2) The electrochemiluminescent marker is terpyridine ruthenium or quantum dots;

[0052] (3) The chemiluminescent marker is a quantum dot;

[0053] (4) before using the device for electrochemiluminescence detection, including performing an electrode activation operation, during which an electrochemiluminescence co-reaction reagent is introduced into the flow channel;

[0054] (5) The electrochemiluminescent co-reaction reagent contains a surfactant for washing the magnetic beads, the surfactant is selected from at least one of Tween, Triton and polycatalpol, and the mass percentage of the surfactant is 0.05% to 1%;

[0055] (6) The neutral cleaning solution is a solution containing phosphate and a detergent, wherein the concentration of the phosphate is 10 mM to 500 mM, and the mass percentage of the detergent is 0.05% to 1%.

[0056] In an optional embodiment, the electrochemiluminescent marker is terpyridine ruthenium, and the electrochemiluminescent co-reactant corresponding to terpyridine ruthenium is a solution containing tripropylamine;

[0057] In an optional embodiment, the solution containing tripropylamine is a phosphate solution of tripropylamine;

[0058] In an optional embodiment, the concentration of tripropylamine in the phosphate solution of tripropylamine is 0.1M to 0.2M, and / or the concentration of phosphate in the phosphate solution of tripropylamine is 10mM to 500mM;

[0059] In an optional embodiment, the concentration of terpyridine ruthenium in the phosphate solution of tripropylamine is 0.01 nM to 5 μM.

[0060] In an optional embodiment, the chemiluminescent marker is a quantum dot, and the chemiluminescent co-reaction reagent corresponding to the quantum dot is a solution containing potassium permanganate.

[0061] In an optional embodiment, the solution containing potassium permanganate is a phosphate solution of potassium permanganate;

[0062] In an optional embodiment, the concentration of potassium permanganate in the potassium permanganate phosphate solution is 1 mM to 3 mM, and / or the concentration of phosphate is 1 mM to 100 mM;

[0063] In an optional embodiment, the concentration of the quantum dots in the phosphate solution containing 0.5% Tween 20 by mass is 0.5 μM to 1.5 μM.

[0064] In an optional embodiment, the first binding molecule is a quantum dot-labeled TSH antibody, and the chemiluminescent co-reaction reagent is a solution containing potassium permanganate;

[0065] The second binding molecule is a Tg antibody labeled with terpyridine ruthenium, and the electrochemiluminescence co-reaction reagent is a solution containing tripropylamine.

[0066] In an optional embodiment, both the chemiluminescent marker and the electrochemiluminescent marker are quantum dots;

[0067] The chemiluminescent co-reaction reagent is a solution containing potassium permanganate; the electrochemiluminescent co-reaction reagent is a solution containing tripropylamine;

[0068] Optionally, the quantum dots are CdTe, CdSe, or CuInS 2 @ZnS.

[0069] In an optional embodiment, the first reagent includes 0.05 mg / L to 50.0 mg / L of a first antibody to be detected labeled with quantum dots, 1 mmol / L to 100 mmol / L of a buffered saline solution, 0.5 g / L to 50 g / L of a surfactant, and 0.05 g / L to 5 g / L of a preservative, and the pH value of the first reagent is 5.0 to 9.0;

[0070] The second reagent comprises 0.05 mg / L to 50.0 mg / L of a second antibody to be detected labeled with terpyridine ruthenium, 1 mmol / L to 100 mmol / L of a buffered saline solution, 0.5 g / L to 50 g / L of a surfactant, and 0.05 g / L to 5 g / L of a preservative, and the pH value of the first reagent is 5.0 to 9.0;

[0071] Alternatively, the second reagent includes 0.05 mg / L to 50.0 mg / L of a second antibody to be detected labeled with quantum dots, 1 mmol / L to 100 mmol / L of a buffered saline solution, 0.5 g / L to 50 g / L of a surfactant, and 0.05 g / L to 5 g / L of a preservative, and the pH value of the first reagent is 5.0 to 9.0. The buffered saline solution includes a combination of one or more of PBS (phosphate buffered saline), PB (phosphate), PIPES (piperazine-1,4-diethanesulfonic acid), MES (2-(N-morpholinyl)ethanesulfonic acid), HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid), TRIS (trishydroxymethylaminomethane), HEPPS (4-(2-hydroxyethyl)-1-piperazinepropanesulfonic acid) and Bis-Tris (bis(2-hydroxyethyl)amino-tris(hydroxymethyl)methane).

[0072] In an optional embodiment, the surfactant includes a combination of one or more of polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monooleate, polyethylene glycol tert-octylphenyl ether, polyethylene glycol nonylphenyl ether and fatty alcohol polyoxyethylene ether;

[0073] The preservative includes one or more combinations of ProClin-300, ProClin-950, gentamicin and BND (5-bromo-5-nitro-1,3-dioxane).

[0074] In an optional embodiment, the first object to be detected and the second object to be detected are respectively selected from PCT (procalcitonin), TSH (thyroid stimulating hormone), HCG (human chorionic gonadotropin), CK-MB (creatine kinase isoenzyme), MYO (myoglobin), NT-proBNP (N-terminal pro-brain natriuretic peptide), Tg (thyroglobulin), CRP (C-reactive protein), PGI (pepsinogen I), PGII (pepsinogen II), cTnI (troponin I), cTnT (troponin T), CT (calcitonin), PTH (parathyroid hormone), Ferritin (ferritin), PIIINP (type III procollagen N-terminal peptide), CIV (type IV collagen), LN (laminin), PRL (prolactin), FSH (follicle-stimulating hormone), LH (luteinizing hormone), TBG (thyroid binding globulin), C-peptide, Insulin, H-FABP (heart-type fatty acid binding protein), Lp-PLA2 (lipoprotein-associated phospholipase A2), IL-6 (interleukin-6), HA (hyaluronic acid), ProGRP (progastrin-releasing peptide), AMH (anti-Muller hormone), D-dimer (DD dimer), AFP (alpha-fetoprotein), CEA (carcinoembryonic antigen), SAA (serum amyloid A), CA125 (carbohydrate antigen 125), CA19-9 (carbohydrate antigen 19-9), SCCA (squamous cell carcinoma-associated antigen), NSE (neuron-specific enolase), CYFRA 21-1 (non-small cell lung cancer associated antigen), CA15-3 (carbohydrate antigen 15-3), CA72-4 (carbohydrate antigen 72-4), HE4 (human epididymis protein), PSA (prostate specific antigen), HBsAg (hepatitis B virus surface antigen), HIVAg (human immunodeficiency virus antigen), IGFBP-3 (insulin-like growth factor binding protein-3), Renin (renin), ACTH (adrenocorticotropic hormone), BNP (B-type natriuretic peptide), CA50 (carbohydrate antigen 50), CA24 Any two of IL-2 (carbohydrate antigen 242), S100 (S100 protein), MPO (myeloperoxidase), hGH (human growth hormone), SHBG (sex hormone binding globulin), TNF-α (tumor necrosis factor-α), IL-1β (interleukin-1β), IL-2 (interleukin-2), IL-2R (interleukin-2 receptor), IL-4 (interleukin-4), IL-5 (interleukin-5), IL-8 (interleukin-8), IL-10 (interleukin-10) and IL-17 (interleukin-17).

[0075] The beneficial effects of the embodiments of the present invention include, for example:

[0076] The chemiluminescence-electrochemical luminescence co-detection device provided by the present invention can realize chemiluminescence and electrochemical luminescence in the same device through the coordinated design of a reaction cup, multiple reagent holding containers, a detection needle, an injection control module, a measuring cell, an electrode assembly, a magnetic adsorption component and at least one set of optical detection modules; the design of the chemiluminescence-electrochemical luminescence co-detection device of the present invention greatly improves the convenience of detection when there are multiple substances in the sample to be tested that need to be analyzed and detected, and both luminescence detection and electrochemical luminescence detection are required; and chemiluminescence detection emits light in the flow channel, which can reduce individual differences in testing caused by sample containers compared to conventional chemiluminescence detection, and its accuracy is also higher.

[0077] The chemiluminescence detection method provided by the present invention can realize chemiluminescence and electrochemiluminescence detection at different time points in the measuring cell of the same electrochemical detection device. The detection method is highly convenient and accurate. BRIEF DESCRIPTION OF THE DRAWINGS

[0078] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0079] Figure 1 A diagram of the structure of an apparatus for implementing the method provided by the present invention;

[0080] Figure 2 A flow chart of the first reagent in the one-step combined test provided by the present invention being introduced into the flow channel;

[0081] Figure 3 A flow chart of the first reagent in the two-step combined test provided by the present invention being introduced into the flow channel;

[0082] Figure 4 A timing control diagram of an embodiment of the present invention;

[0083] Figure 5 The graphs are TSH chemiluminescent signal-concentration relationship curves and Tg electrochemiluminescent signal-concentration relationship curves of various examples.

[0084] Icons: 101-reagent container; 102-detection needle; 103-optical detection module; 104-magnetic adsorbent; 105-measuring cell; 105a-flow channel; 106-reaction cup; 107-filter; 108-receiving tank; A-chemiluminescent co-reaction reagent; B-electrochemiluminescent co-reaction reagent; C-alkaline cleaning solution; D-neutral cleaning solution; S-sample mixture. DETAILED DESCRIPTION

[0085] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Generally, the components of the embodiments of the present invention described and shown in the drawings here can be arranged and designed in various different configurations.

[0086] Therefore, the following detailed description of the embodiments of the present invention provided in the accompanying drawings is not intended to limit the scope of the invention claimed for protection, but merely represents selected embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0087] It should be noted that similar reference numerals and letters denote similar items in the following drawings, and therefore, once an item is defined in one drawing, further definition and explanation thereof is not required in subsequent drawings.

[0088] In the description of the present invention, it should be noted that if the terms "upper", "lower", "inside", "outside", etc. appear to indicate an orientation or position relationship, they are based on the orientation or position relationship shown in the accompanying drawings, or are the orientation or position relationship in which the product of the invention is usually placed when used. They are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present invention.

[0089] In addition, the terms “first”, “second”, etc., if used, are merely used to distinguish between the descriptions and should not be understood as indicating or implying relative importance.

[0090] It should be noted that, in the absence of conflict, the features in the embodiments of the present invention may be combined with each other.

[0091] Please refer to Figure 1 , Figure 1 The chemiluminescence co-detection device provided by an embodiment of the present invention includes:

[0092] Reaction cup 106, multiple reagent containers 101, detection needle 102, injection control module, measurement pool 105, electrode assembly, magnetic adsorption element 104 and at least one set of optical detection module 103;

[0093] The reaction cup 106 is used to hold a sample mixture, which includes a conjugate; the conjugate is obtained by specifically binding at least two analytes in the sample to be tested to at least two binding molecules in a one-to-one correspondence and adsorbing them onto magnetic beads; at least one of the at least two binding molecules is a molecule labeled with a chemiluminescent marker, and at least one of the at least two binding molecules is a molecule labeled with an electrochemiluminescent marker;

[0094] At least a portion of the multiple reagent holding containers 101 are used to hold multiple luminescent reagents, respectively, and the multiple luminescent reagents include at least one chemiluminescent co-reactant and at least one electrochemiluminescent co-reactant;

[0095] The detection needle 102 is used to draw the sample mixture from the reaction cup 106 into the flow channel 105a of the measuring cell 105, and is used to draw the corresponding luminescent reagents from the multiple reagent holding containers 101 into the flow channel 105a of the measuring cell 105;

[0096] The sample injection control module is used to control the detection needle 102 to absorb into the reaction cup 106 and the multiple reagent holding containers 101;

[0097] The measuring cell 105 is used to provide a reaction space for realizing chemiluminescence and electrochemiluminescence;

[0098] The electrode assembly is arranged in conjunction with the measuring cell 105, and the electrode assembly is used to excite the electrochemical luminescent marker to emit light after being energized;

[0099] The magnetic adsorption member 104 is disposed close to the measuring cell 105 and is used to magnetically attract the magnetic beads in the flow channel 105a entering the measuring cell 105;

[0100] At least one set of optical detection modules 103 is used to detect and collect the luminous intensity of the luminescent marker;

[0101] The chemiluminescence co-detection device is configured as follows:

[0102] The sampling control module controls the detection needle 102 to absorb the sample mixture into the flow channel 105a of the measuring cell 105;

[0103] After the object to be bound is adsorbed by the magnetic adsorbent 104, the injection control module controls the detection needle 102 to absorb the luminescent reagent into the flow channel 105a of the measuring pool 105. When the absorbed luminescent reagent is a chemiluminescent co-reactant, the chemiluminescent co-reactant combines with the corresponding luminescent marker to realize chemiluminescence, and the luminescent intensity is detected and collected by at least one set of optical detection modules 103;

[0104] When the absorbed luminescent reagent is an electrochemiluminescent co-reactant, the electrode assembly is energized, the electrochemiluminescent co-reactant combines with the corresponding luminescent marker to realize electrochemiluminescence, and the luminescent intensity is detected and collected by at least one set of optical detection modules 103 .

[0105] The chemiluminescence-electrochemiluminescence co-detection device provided by the present invention is suitable for performing chemiluminescence and electrochemiluminescence detection on a sample to be tested having a plurality of analytes. After the binding molecule labeled with the luminescent marker is fully mixed and reacted with the sample to be tested and the magnetic beads, only one injection is required, and the corresponding chemiluminescence co-reaction reagent and electrochemiluminescence co-reaction reagent are subsequently introduced at different time points to realize chemiluminescence detection and electrochemiluminescence detection. The operation is simple and the luminescence accuracy in the flow channel 105a is high.

[0106] It should be noted that, in the co-detection device provided by the present invention, the number of chemiluminescence detections can be one or more times during the entire luminescence detection process. To achieve multiple chemiluminescence detections, it is necessary to use multiple chemiluminescence markers to label different molecules and bind to magnetic beads. Accordingly, the types of chemiluminescence co-reaction reagents introduced into the flow channel 105a are also multiple types that match the chemiluminescence markers.

[0107] Similarly, in the entire luminescence detection process, the number of electrochemiluminescence detections can be one time or multiple times. To achieve multiple electrochemiluminescence detections, a variety of electrochemiluminescence markers are required to label different molecules and combine with magnetic beads. Accordingly, the types of electrochemiluminescence co-reaction reagents introduced into the flow channel 105a are also a variety that match the electrochemiluminescence markers.

[0108] The chemiluminescence detection and electrochemiluminescence detection share the measuring cell 105 as a luminescent container. Both electrochemiluminescence reaction and chemiluminescence reaction can be triggered in the measuring cell 105. Further, the structure of the measuring cell 105 is substantially the same as that of the measuring cell 105 of conventional electrochemiluminescence detection. Specifically, an electrode assembly is provided in conjunction with the measuring cell 105, and the electrode assembly has 2 or 3 electrodes. When the electrode assembly has 3 electrodes, they are a working electrode (WE), an auxiliary electrode (CE) and a reference electrode (RE). When the electrode assembly has 2 electrodes, it does not include a reference electrode (RE). Among them, a platinum electrode is used as a working electrode (WE) and an auxiliary electrode (CE), and a silver chloride electrode is used as a reference electrode (RE). Generally, the working electrode (WE) is located in the flow channel 105a of the measuring cell 105, and is arranged at the bottom of the flow channel 105a, and the auxiliary electrode (CE) and the reference electrode (RE) are arranged on a transparent window of the luminescent region.

[0109] The optical detection module 103 also includes a magnetic adsorbent 104. The magnetic adsorbent 104 is located at the bottom of the measuring cell 105. The magnetic adsorbent 104 can move toward the bottom of the measuring cell 105 and attract the magnetic particles in the measuring cell 105. The magnetic adsorbent 104 can move away from the bottom of the measuring cell 105 to release the attraction of the magnetic particles in the measuring cell 105. Further, the magnetic adsorbent 104 moves by moving up and down relative to the measuring cell 105, and can also move by rotating and swinging relative to the measuring cell 105.

[0110] Specifically, a receiving groove 108 for receiving at least part of the magnetic adsorbent 104 is provided at the bottom of the outer wall of the measuring cell 105. The magnetic adsorbent 104 can move to the receiving groove 108 close to the bottom of the measuring cell 105 to attract the magnetic particles in the measuring cell 105. The provision of the receiving groove 108 facilitates the loading and evacuation of the magnetic adsorbent 104. Specifically, the receiving groove 108 is formed by an inward depression of the bottom of the measuring cell 105. The position of the receiving groove 108 is roughly opposite to the electrode sheet of the light-emitting area, which helps the magnetic particles in the measuring cell 105 to be attracted by the magnetic adsorbent 104 and stay on the surface of the electrode sheet.

[0111] The magnetic adsorption component 104 may be, for example, a permanent magnet or an electromagnet.

[0112] Optionally, when the magnetic adsorbent 104 is a permanent magnet, the chemical-electrochemical luminescence co-inspection equipment also includes a movement control module, which controls the magnetic adsorbent 104 to move into the receiving groove 108 when the luminescence detection starts, and controls the magnetic adsorbent 104 to move out of the receiving groove 108 after the luminescence detection ends to release the adsorption of the magnetic beads by the magnetic adsorbent 104.

[0113] Optionally, when the magnetic adsorption member 104 is an electromagnet, the electromagnet loses its magnetic force after power is turned off, so there is no need to additionally set a motion mechanism for controlling the motion of the electromagnet.

[0114] Furthermore, the number of optical detection modules 103 is one or more sets, which are installed above the light-emitting area window of the measuring cell 105. A filter 107 of a specific wavelength is provided on the sensor receiving window of each set of optical detection modules 103. The filter 107 is used to distinguish the luminous intensity of different luminous objects. Specifically, the bandpass filter wavelength can be selected by switching the filter 107. Among them, the filter 107 is a narrow-band filter lens. According to the wavelength of the luminous object, the corresponding narrow-band filter lens is matched to improve the anti-interference performance of each joint inspection item and avoid mutual influence of each joint inspection item.

[0115] It should be noted that the number of optical detection modules 103 is not limited to multiple sets. If the discrimination of multiple luminescent objects using co-reaction reagents meets the measurement range of each item, the number of optical detection modules 103 can also be one set. Whether one or more sets of optical detection modules 103 are set depends on whether the luminescent wavelength can be received by the photomultiplier tube with high sensitivity.

[0116] Optionally, each optical detection module 103 includes a photon detector and a collector; the photon detector is an avalanche diode (APD, a silicon photomultiplier tube (Si-PMT), a photomultiplier tube (PMT) or a photodiode (PD); and / or the collector is a linear amplifier, a logarithmic amplifier, an analog-to-digital conversion circuit, an IV conversion circuit or a photon counter.

[0117] Optionally, the chemi-electrochemical luminescence co-detection device may further include a reaction module, the main function of which is to provide a better reaction environment to promote the test sample, the binding molecule and the magnetic beads to fully react or fully bind. Specifically, the reaction module may be a heater.

[0118] Furthermore, the chemical-electrochemical luminescence co-detection device also includes a control system;

[0119] The control system is in communication with the injection control module and the electrode assembly. The control system has a built-in timing control program. The timing control program is configured to instruct the injection control module to control the detection needle to absorb the corresponding reagent from different reagent holding containers 101 at different time points.

[0120] When the current time node is the time node for absorbing the electrochemiluminescent co-reaction reagent, the control system sends a command to the electrode assembly to power it on. After the electrochemiluminescent reaction is completed, the control system controls the electrode assembly to power off.

[0121] The setting of the control system and the timing control program can realize the automatic sampling of chemiluminescence and electrochemiluminescence at specific time nodes, and control the power supply of the electrode assembly at the appropriate time nodes, thereby realizing the automatic detection of chemiluminescence and electrochemiluminescence.

[0122] In addition, the control system is also connected to the optical detection module 103 for communication. The optical detection module 103 feeds back the detected light intensity information to the control system, and the control system converts the light intensity information into the concentration information of the object being measured.

[0123] Optionally, the timing control program is configured to: control the sampling control module to make the detection needle 102 first sample the chemiluminescent co-reaction reagent to achieve chemiluminescence, and then sample the electrochemiluminescent co-reaction reagent to achieve electrochemiluminescence;

[0124] Alternatively, the timing control program is configured to: control the injection control module to make the detection needle 102 first sample the electrochemiluminescent co-reaction reagent to achieve electrochemiluminescence, and then sample the chemiluminescent co-reaction reagent to achieve chemiluminescence.

[0125] It should be noted that when chemiluminescence detection is performed multiple times and electrochemiluminescence detection is performed multiple times during the entire detection process, for example, the timing control program can be configured to: sequentially perform chemiluminescence co-reaction reagent sampling, electrochemiluminescence co-reaction reagent sampling, chemiluminescence co-reaction reagent sampling, electrochemiluminescence co-reaction reagent sampling, etc.; it can also be electrochemiluminescence co-reaction reagent sampling, electrochemiluminescence co-reaction reagent sampling, chemiluminescence co-reaction reagent sampling, chemiluminescence co-reaction reagent sampling, etc.

[0126] Optionally, some of the multiple reagent holding containers 101 are used to hold a neutral cleaning solution;

[0127] The timing control program is configured to: manipulate the sampling control module to make the detection needle 102 perform sampling of the sample mixture, absorption of the neutral cleaning solution, sampling of the chemiluminescent co-reaction reagent, and sampling of the electrochemiluminescent co-reaction reagent in sequence;

[0128] Alternatively, the timing control program is configured to: manipulate the injection control module to cause the detection needle 102 to sequentially perform sample mixture sampling, neutral cleaning solution aspiration, electrochemiluminescence co-reaction reagent sampling, and chemiluminescence co-reaction reagent sampling.

[0129] That is, the timing control program is configured to: enable the detection needle 102 to absorb the neutral cleaning solution after the sampling of the sample mixture is completed, and to absorb the neutral cleaning solution between the sampling of the two luminescent reagents.

[0130] Optionally, the detection needle 102 is also used to draw air into the flow channel 105a of the measuring cell 105 to form bubbles in the flow channel 105a;

[0131] The timing control program is configured to: manipulate the sampling control module to make the detection needle 102 sequentially perform sample mixture sampling, neutral cleaning solution aspiration, air aspiration, chemiluminescent co-reaction reagent sampling, and electrochemiluminescent co-reaction reagent sampling;

[0132] The timing control program is configured to: manipulate the injection control module to make the detection needle 102 sequentially perform sample mixture sampling, neutral cleaning solution aspiration, air aspiration, electrochemiluminescence co-reaction reagent sampling, and chemiluminescence co-reaction reagent sampling.

[0133] That is, the timing control program is configured to: after the neutral cleaning solution is sucked out, make the detection needle 102 suck out air to isolate the neutral cleaning solution and the luminescent reagent introduced subsequently.

[0134] The present invention provides a chemiluminescence detection method, comprising the following steps:

[0135] A sample mixture is introduced into the flow channel 105a of the measuring cell 105. The sample mixture includes a conjugate, which is mainly obtained by fully reacting the sample to be tested, at least one first binding molecule, at least one second binding molecule and magnetic beads. The conjugate is adsorbed on the bottom of the flow channel 105a when passing near the magnetic adsorbent 104 disposed outside the flow channel 105a.

[0136] A plurality of luminescent reagents are introduced into the flow channel 105a of the measuring cell 105, each luminescent reagent is introduced at a different time point, and the plurality of luminescent reagents include at least one chemiluminescent co-reaction reagent and at least one electrochemiluminescent co-reaction reagent;

[0137] Each first binding molecule is a molecule labeled with a chemiluminescent marker. When there are multiple first binding molecules in the sample mixture, the multiple first binding molecules are molecules labeled with different chemiluminescent markers, and each first binding molecule is a substance that can specifically bind to a corresponding analyte in the sample to be tested and adsorb to the magnetic beads;

[0138] Each second binding molecule is a molecule labeled with an electrochemiluminescent marker. When the number of second binding molecules in the sample mixture is multiple, the multiple second binding molecules are molecules labeled with different electrochemiluminescent markers, and each second binding molecule is a substance that can specifically bind to a corresponding analyte in the sample to be tested and adsorbed to the magnetic beads;

[0139] After the at least one chemiluminescent co-reaction reagent is bound to the luminescent marker on the at least one first binding molecule in a one-to-one correspondence, chemiluminescence is realized, and the corresponding luminescence intensity is detected and collected by the optical detection module 103 of the device;

[0140] When the electrochemiluminescent co-reaction reagent is introduced into the flow channel 105a, the electrode assembly in the measuring cell 105 is energized, and at least one electrochemiluminescent co-reaction reagent is combined with the luminescent marker on at least one second binding molecule in a one-to-one correspondence to realize electrochemiluminescence, and the corresponding luminescence intensity is detected and collected by the optical detection module 103 of the device.

[0141] In the detection method provided by the present invention, after the sample is injected into the flow channel 105a, the magnetic beads are adsorbed by the magnetic adsorbent 104 at the bottom of the flow channel 105a in the measuring cell 105, and then the luminescent reagent (chemiluminescent co-reaction reagent, electrochemiluminescent co-reaction reagent) can be introduced into the flow channel 105a for luminescent detection. The method provided by the present invention can realize electrochemiluminescent detection and chemiluminescent detection in the same device. When the sample requires both luminescent detection and electrochemiluminescent detection, the convenience of detection is greatly improved; and the chemiluminescent detection in the flow channel 105a can reduce the individual differences in the test caused by the sample container compared to the conventional chemiluminescent detection, and its accuracy is also higher. The detection method provided by the present invention can be implemented by the device provided in the embodiment of the present invention, and can also be implemented by the existing electrochemiluminescent detection equipment.

[0142] It should be noted that the detection method provided by the present invention can detect multiple substances in the sample to be tested, and can perform at least one chemiluminescence detection and at least one electrochemiluminescence detection. For performing several chemiluminescence detections, there are corresponding several chemiluminescence markers, corresponding several first binding molecules, and corresponding several additions of chemiluminescence co-reaction reagents (the chemiluminescence co-reaction reagents added multiple times can be the same or different, and the principle of addition is that they can be combined with the chemiluminescence marker to emit light); for performing several electrochemiluminescence detections, there are corresponding several electrochemiluminescence markers, corresponding several second binding molecules, and corresponding several additions of electrochemiluminescence co-reaction reagents (the electrochemiluminescence co-reaction reagents added multiple times can be the same or different, and the principle of addition is that they can be combined with the electrochemiluminescence marker to emit light).

[0143] Specifically, taking a chemiluminescence detection and an electrochemiluminescence detection as an example, the specific method of performing the chemiluminescence detection and the electrochemiluminescence detection in the same electrochemiluminescence detection device is as follows:

[0144] 1. The first type (one-step combined test: sample and test reagent are introduced at one time):

[0145] First, chemiluminescence detection is performed, and then electrochemiluminescence detection is performed. The specific detection method is as follows:

[0146] S1. Sample mixing reaction

[0147] The sample to be tested, at least one first reagent and at least one second reagent are placed in a reaction cup 106 and mixed evenly, and then placed in a reaction module for full reaction, and then streptavidin-labeled magnetic beads are added, mixed evenly, and then placed in the reaction module for full reaction. Electrochemical luminescence usually uses magnetic beads with a particle size of 1 μm to 3.0 μm, balancing the loss rate of magnetic beads and the cleaning effect of the measuring pool 105. After the reaction is completed, a sample mixture is obtained.

[0148] The first reagent is a first binding molecule labeled with a chemiluminescent marker, and the first binding molecule is a substance that can specifically bind to the first analyte in the sample to be tested; the second reagent is a second binding molecule labeled with an electrochemiluminescent marker, and the second binding molecule is a substance that can specifically bind to the second analyte in the sample to be tested and adsorbed to the magnetic beads.

[0149] In this step, the test sample, the first reagent, the second reagent and the magnetic beads react fully, so that the first binding molecule labeled with the chemiluminescent marker and the first analyte bind to the magnetic beads, and the second binding molecule labeled with the electrochemiluminescent marker and the second analyte bind to the magnetic beads to form a complex.

[0150] Optionally, the chemiluminescent marker is a quantum dot, and the chemiluminescent co-reaction reagent (reagent A) corresponding to the quantum dot is a solution containing potassium permanganate. Further, the solution containing potassium permanganate is a phosphate solution of potassium permanganate. The concentration of potassium permanganate in the phosphate solution of potassium permanganate is 1mM to 3mM, and / or the concentration of phosphate is 1mM to 100mM. The concentration of quantum dots in a phosphate solution containing 0.5% Tween 20 by mass is 0.5μM to 1.5μM, at which the concentration of quantum dots can be guaranteed to be equivalent to electrochemiluminescence.

[0151] Optionally, the electrochemiluminescent marker is terpyridine ruthenium, and the electrochemiluminescent co-reaction reagent (reagent B) corresponding to terpyridine ruthenium is a solution containing tripropylamine. Further, the solution containing tripropylamine is a phosphate solution of tripropylamine. The concentration of tripropylamine in the phosphate solution of tripropylamine is 0.1M to 0.2M, and / or the concentration of phosphate in the phosphate solution of tripropylamine is 10mM to 500mM. To ensure the luminescence intensity, the concentration of terpyridine ruthenium in the phosphate solution of tripropylamine is 0.01nM to 5μM.

[0152] Particularly preferably, the chemiluminescent marker and the electrochemiluminescent marker are both quantum dots; optionally, the quantum dots are CdTe, CdSe or CuInS 2 @ZnS. It should be noted that when both the chemiluminescent marker and the electrochemiluminescent marker are quantum dots, the same quantum dots or different quantum dots can be selected. When the same quantum dots are selected, the same quantum dots can detect two analytes in one measuring cell 105 through chemiluminescence and electrochemiluminescence; when different quantum dots are selected, multiple quantum dots can achieve time- and wavelength-resolved multiple detections through chemiluminescence and electrochemiluminescence.

[0153] Optionally, the chemiluminescent co-reactant is a solution containing potassium permanganate; and the electrochemiluminescent co-reactant is a solution containing tripropylamine.

[0154] Optionally, the first reagent includes quantum dot-labeled TSH antibody 0.5-5.0 μg / mL (e.g., 0.5 μg / mL, 1 μg / mL, 3 μg / mL, or 5 μg / mL), PBS (phosphate buffered saline) 0.01M-0.1M (e.g., 0.01M, 0.02M, 0.05M, or 0.1M), Tween-20 0.05-1%wt (e.g., 0.05%wt, 0.1%wt, 0.2%wt, 0.5%wt, or 1%wt), Triton X-100 0.05~1%wt (for example 0.05%wt, 0.1%wt, 0.2%wt, 0.5%wt or 1%wt), BSA (bovine serum albumin) 0.5~5%wt (for example 0.5%wt, 0.8%wt, 1%wt, 3%wt or 5%wt), ProClin-3000.05~0.5%wt (for example 0.05%wt, 0.1%wt, 0.2%wt or 0.5%wt), the pH value of the first reagent is 7.3~7.5 (for example 7.3, 7.4 or 7.5).

[0155] The role of PBS in the component is to provide a buffer environment, which can ensure pH stability within the above concentration range. If the concentration is too high, it will cause protein denaturation; the role of Tween-20 and Triton X-100 in the component is to protect the higher-level structure of the protein, which can ensure the antibody remains active and stable within the above concentration range. If the concentration is too high, it will cause protein inactivation; the role of BSA in the component is to block the nonspecific binding sites in the reaction, which can reduce nonspecific adsorption interference within the above concentration range. If the concentration is too high, it will cause a decrease in the luminescent signal; the role of ProClin-300 in the component is antibacterial and preservative, which can ensure the long-term stability of the solution within the above concentration range. If the concentration is too high, it will cause a decrease in the luminescent signal. The pH range in the first reagent is the optimal buffer range of PBS.

[0156] Optionally, the second reagent includes terpyridine ruthenium labeled Tg antibody 0.1-5.0 μg / mL (e.g., 0.1 μg / mL, 1 μg / mL, 3 μg / mL or 5 μg / mL), MES (2-(N-morpholino)ethanesulfonic acid) 0.02M-0.2M (e.g., 0.02M, 0.05M, 0.1M or 0.2M), Tween-20 0.05-1%wt, e.g., 0.05%wt, 0.1%wt, 0.2%wt, 0.5%wt or 1%wt), TritonX-100 0.05~1%wt (for example 0.05%wt, 0.1%wt, 0.2%wt, 0.5%wt or 1%wt), BSA (bovine serum albumin) 0.5~5%wt (for example 0.5%wt, 0.8%wt, 1%wt, 3%wt or 5%wt), BND (5-bromo-5-nitro-1,3-dioxane) 0.05~0.5%wt (for example 0.05%wt, 0.1%wt, 0.2%wt or 0.5%wt), the pH value of the second reagent is 5.9~6.1 (for example 5.9, 6 or 6.1).

[0157] The role of MES in the component is to provide a buffer environment, which can ensure pH stability within the above concentration range. If the concentration is too high, it will cause protein denaturation; the role of Tween-20 and Triton X-100 in the component is to protect the higher-level structure of the protein, which can ensure the antibody remains active and stable within the above concentration range. If the concentration is too high, it will cause protein inactivation; the role of BSA in the component is to block the nonspecific binding sites in the reaction, which can reduce nonspecific adsorption interference within the above concentration range. If the concentration is too high, it will cause the luminescence signal to decrease; the role of BND in the component is antibacterial and preservative, which can ensure the long-term stability of the solution within the above concentration range. If the concentration is too high, it will cause the luminescence signal to decrease. The pH range in the first reagent is the optimal buffer range of MES.

[0158] Alternatively, the second reagent includes quantum dot-labeled Tg antibody 0.1-5.0 μg / mL (e.g., 0.1 μg / mL, 0.5 μg / mL, 1 μg / mL, 3 μg / mL, or 5 μg / mL), PBS 0.01M-0.1M (e.g., 0.01M, 0.02M, 0.05M, or 0.1M), Tween-20 0.05-1%wt (e.g., 0.05%wt, 0.1%wt, 0.2%wt, 0.5%wt, or 1%wt), Triton X-100 0.05-1%wt (e.g., 0.05%wt, 0.1%wt, 0.2%wt, 0.5%wt, or 1%wt), BSA 0.5-5%wt (e.g., 0.5%wt, 0.8%wt, 1%wt, 3%wt, or 5%wt), ProClin-300 0.05-0.5% wt (eg, 0.05% wt, 0.1% wt, 0.2% wt or 0.5% wt), and the pH value of the second reagent is 7.3-7.5 (eg, 7.3, 7.4 or 7.5).

[0159] S2, reagents are introduced into flow channel 105a in sequence

[0160] Please refer to the specific process Figure 2 As shown, after the electrochemiluminescence detection device is started, according to the timing control, the electrochemiluminescence co-reaction reagent (reagent B) is first sucked into the flow channel 105a by the detection needle 102, and the electrode is energized to achieve electrode activation;

[0161] Then, the sample mixture (S) is sucked by the detection needle 102. When the sample mixture is left in the measuring pool 105, the magnet under the measuring pool 105 moves and is closely attached to the receiving groove 108 below the luminous area at the bottom of the measuring pool 105. The conjugate slowly flows through the reaction flow channel 105a of the measuring pool 105 under the timing control, and the magnetic beads in the reaction conjugate are adsorbed to the bottom of the flow channel 105a above the magnetic adsorbent 104. The flow rate is usually 10μL / S to 30μL / S, and the flow channel 105a at the magnetic separation position of the measuring pool 105 is flat to ensure the effective magnetic bead adsorption area and cleaning effect.

[0162] Then, the neutral cleaning fluid (reagent D) is sucked through the detection needle 102 and passed into the flow channel 105a to repeatedly clean the magnetic beads, and the substances not bound to the magnetic beads are taken away by the reagent D under the action of the fluid.

[0163] Then, bubbles sufficient to cover the flow channel 105 a are sucked into the flow channel 105 a through the detection needle 102 to separate the reagent D and the reagent A. For example, 10 μL to 50 μL of bubbles are used for isolation.

[0164] Then, reagent A is sucked into the flow channel 105a using the detection needle 102, for example, at a flow rate of 100 μL / S to 500 μL / speed through the measuring cell 105, to ensure that reagent A is quickly combined with the magnetic beads combined with quantum dots, is not diluted by the residual reagent D, and does not wash away the magnetic beads. When reagent A reaches the magnetic bead area of ​​the measuring cell 105, the quantum dots emit light; the electrochemical measuring cell 105 is not powered during the process of quantum dots adsorbing to the magnetic beads, the process of magnetic bead cleaning, and the process of luminescence. The light emission of quantum dots is detected by the photon detection and collection device 103; usually, there is a focusing lens and a spectroscopic lens between the photon detection and collection device 103 and the measuring cell 105.

[0165] After the quantum dots have finished emitting light, the detection needle 102 is used to absorb the electrochemiluminescent co-reaction reagent (reagent B) with a magnetic bead cleaning effect into the flow channel 105a. Reagent B does not react with quantum dots and terpyridine ruthenium when it is not powered on. During the process of reagent B passing into the flow channel 105a, the magnetic adsorption member 104 is close to the bottom of the measuring cell 105 to maintain the adsorption of the magnetic beads. The reagent filling is completed at a speed of 100μL / S to 200μL / S. After the magnetic beads are cleaned, the measuring cell 105 is powered on. If the electrochemiluminescent marker is terpyridine ruthenium, 1.4V can be used to excite electrochemiluminescence (terpyridine ruthenium and tripropylamine usually have an excitation voltage of 1.2V to 1.5V). After the terpyridine ruthenium emits light, it is detected by the photon detection and collection device 103.

[0166] Optionally, in this step, before introducing reagent B, a neutral reagent (reagent D) may be introduced for cleaning.

[0167] After the luminescence is completed, similar to the common electrochemical luminescence measuring cell 105, an alkaline cleaning solution (reagent C) is introduced into the flow channel 105a, so that the measuring cell 105, the detection needle 102, and the pipeline are cleaned by the reagent C. During the cleaning process, a step-change voltage is applied to ensure the cleaning effect. The applied voltage is different according to the different electrode materials of the electrochemical measuring cell 105. After the cleaning is completed, the power is turned on to restore the electrode surface and re-fill the measuring cell 105.

[0168] Optionally, the electrochemiluminescent co-reaction reagent (reagent B) contains a surfactant for washing the magnetic beads, the surfactant is selected from at least one of Tween, Triton and polycatalpol, and the mass percentage of the surfactant is 0.05% to 1%.

[0169] Optionally, the alkaline cleaning solution (reagent C) is a solution containing a detergent and a strong base, wherein the mass percentage of the detergent is 0.05% to 1%, the detergent is selected from at least one of Tween, Triton and polycatalpol, the strong base is selected from at least one of KOH and NaOH, and the pH of the alkaline cleaning solution is 12 to 14.

[0170] Optionally, the neutral cleaning solution (reagent D) is a solution containing phosphate and a detergent, the concentration of the phosphate is 10 mM to 500 mM, and the mass percentage of the detergent is 0.05% to 1%.

[0171] 2. The second type (one-step combined test: sample and test reagent are introduced at one time):

[0172] Electrochemiluminescence detection is performed first and then chemiluminescence detection. The specific detection method is as follows:

[0173] The reagents and some cleaning operations involved in this detection method refer to the specific contents described in the first method, and will not be described in detail here.

[0174] S1. Sample mixing reaction

[0175] The sample to be tested, the first reagent and the second reagent are placed in the reaction cup 106 and mixed evenly, and then the streptavidin-labeled magnetic beads are added, mixed evenly and then placed in the reaction module for full reaction. After the reaction is completed, a sample mixture is obtained.

[0176] S2, reagents are introduced into flow channel 105a in sequence

[0177] In this detection method, the order of reagent introduction is:

[0178] Reagent B is introduced to activate the electrode by electrifying the electrode, the sample mixture is introduced, reagent D is introduced to clean the flow channel 105a and the magnetic beads, electrochemiluminescence detection is performed using reagent B, reagent D is introduced to clean the flow channel 105a and the magnetic beads, bubbles are introduced to isolate reagent D and reagent A, and reagent A is introduced to perform chemiluminescence detection.

[0179] In addition to the above two methods, the method provided by the present invention can also perform the following two tests using the device provided by the present invention. The following two tests are two-step sample addition, which is slightly less convenient than the one-step combined test and one-step sample addition provided by the present invention.

[0180] 1. The first type (two-step joint test: samples and test reagents are introduced in batches):

[0181] First, chemiluminescence detection is performed, and then electrochemiluminescence detection is performed. The specific detection method is as follows:

[0182] The reagents and some cleaning operations involved in this detection method refer to the specific contents described in the first one-step combined test, and will not be described in detail here.

[0183] S1. Sample mixing reaction

[0184] The sample to be tested and the first reagent are placed in the reaction cup 106 and mixed evenly, and then placed in the reaction module for full reaction, and then streptavidin-labeled magnetic beads are added, mixed evenly, and then placed in the reaction module for full reaction. After the reaction is completed, a chemiluminescent mixed sample is obtained, in which the first binding molecule and the first analyte are bound to the magnetic beads.

[0185] The sample to be tested and the second reagent are placed in the reaction cup 106 and mixed evenly, and then placed in the reaction module for full reaction, and then streptavidin-labeled magnetic beads are added, mixed evenly, and then placed in the reaction module for full reaction. After the reaction is completed, an electrochemical luminescent mixed sample is obtained, and the second binding molecule and the second analyte are bound to the magnetic beads.

[0186] S2, reagents are introduced into flow channel 105a in sequence

[0187] Please refer to the specific process Figure 2 As shown, in this detection method, the order of reagent introduction is:

[0188] Pass reagent B to activate the electrode by electrifying the electrode, pass the chemiluminescent sample mixture, pass reagent D to clean the flow channel 105a and the magnetic beads, pass the bubble isolation reagent D and reagent A, pass reagent A to perform chemiluminescent detection, pass reagent D to clean the flow channel 105a, pass the electrochemiluminescent sample mixture, pass reagent B to clean the magnetic beads and perform electrochemiluminescent detection.

[0189] 2. The second type (two-step joint test: samples and test reagents are introduced in batches):

[0190] Electrochemiluminescence detection is performed first and then chemiluminescence detection. The specific detection method is as follows:

[0191] The reagents and some cleaning operations involved in this detection method refer to the specific contents described in the first one-step combined test, and will not be described in detail here.

[0192] S1. Sample mixing reaction

[0193] The sample to be tested and the first reagent are placed in the reaction cup 106 and mixed evenly, and then placed in the reaction module for full reaction, and then streptavidin-labeled magnetic beads are added, mixed evenly, and then placed in the reaction module for full reaction. After the reaction is completed, a chemiluminescent mixed sample is obtained, in which the first binding molecule and the first analyte are bound to the magnetic beads.

[0194] The sample to be tested and the second reagent are placed in the reaction cup 106 and mixed evenly, and then placed in the reaction module for full reaction, and then streptavidin-labeled magnetic beads are added, mixed evenly, and then placed in the reaction module for full reaction. After the reaction is completed, an electrochemical luminescent mixed sample is obtained, and the second binding molecule and the second analyte are bound to the magnetic beads.

[0195] S2, reagents are introduced into flow channel 105a in sequence

[0196] In this detection method, the order of reagent introduction is:

[0197] Pass reagent B, energize the electrodes to activate them, pass an electrochemiluminescent mixed sample, pass reagent D to clean the flow channel 105a and the magnetic beads, pass reagent B for electrochemiluminescence detection, pass a chemiluminescent mixed sample, pass reagent D to clean the flow channel 105a and the magnetic beads, pass bubbles to isolate reagent D and reagent A, and pass reagent A for chemiluminescence detection.

[0198] The following describes the invention in conjunction with specific embodiments.

[0199] The embodiments of the present application will be described in detail below in conjunction with examples. It should be understood that these examples are only used to illustrate the present application and are not intended to limit the scope of the present application. The experimental methods for which specific conditions are not specified in the following examples are preferably referred to the guidance provided in the present application, and can also be based on the experimental manual or normal conditions in this area, can also be based on the conditions recommended by the manufacturer, or refer to experimental methods known in the art.

[0200] In the following specific embodiments, the measured parameters of raw material components may have slight deviations within the range of weighing accuracy unless otherwise specified. For temperature and time parameters, acceptable deviations caused by instrument test accuracy or operation accuracy are allowed.

[0201] Example 1

[0202] In this embodiment, the thyroid stimulating hormone (TSH) and thyroglobulin (Tg) joint test kits of the thyroid function test package are used as an example. First, the components of the TSH and Tg joint test kits include quantum dot-labeled TSH antibody (concentration of 2.0 μg / mL) and terpyridine ruthenium-labeled Tg antibody. The specific components of the first reagent are: quantum dot-labeled TSH antibody 2.0 μg / mL, PBS 0.05M, Tween-20 0.5%wt, Triton X-100 0.5%wt, BSA 3%wt, ProClin-300 0.2%wt, pH value 7.4, and the specific components of the second reagent are: terpyridine ruthenium-labeled Tg antibody 0.5 μg / mL, MES 0.1M, Tween-20 0.3%wt, Triton X-100 0.5%wt, BSA 2%wt, BND 0.3%wt, pH value is 6; the mixture needs to be continuously mixed during the test process due to the presence of quantum dots; like the general sandwich reaction, the mixture, together with biotin-labeled TSH antibody and biotin-labeled Tg antibody, are added to the reaction container (reaction cup 106) by the reagent loading device; the sample loading device adds the sample to be tested to the reaction cup 106 and mixes it, then puts it into the reaction module for reaction for 9 minutes. After the reaction is completed, the reagent loading device adds streptavidin-labeled magnetic beads, mixes them, and then puts them into the reaction module for another 9 minutes of reaction. Among them, the average particle size of the magnetic beads used is 1μm to 3.0μm. The quantum dot-labeled TSH antibody comes from Pumen Technology, the quantum dots are CdTe quantum dots, and the tripyridine ruthenium-labeled Tg antibody comes from Pumen Technology.

[0203] After the reaction is completed, the sample to be tested and the reagent form a conjugate, which is sucked into the measuring pool 105 of the optical detection module 103 by the detection needle 102 through the flow channel 105a. At this time, the magnetic adsorption member 104 at the bottom of the measuring pool 105 moves and sticks to the pit below the luminous area at the bottom of the measuring pool 105. Figure 4 As shown, the conjugate slowly flows through the reaction channel 105a of the measuring cell 105 under timing control, and the magnetic beads in the reaction conjugate are adsorbed to the bottom of the channel 105a above the magnetic adsorbent 104, with a flow rate of 20 μL / S.

[0204] The detection needle 102 absorbs the reagent D and repeatedly washes the magnetic beads, and the substances not bound to the magnetic beads are taken away by the reagent D under the action of the fluid.

[0205] The detection needle 102 draws 30 μL of air bubbles to isolate the reagent D and the reagent A.

[0206] Reagent A is introduced and passed through the measuring cell 105 at a flow rate of 300 μL / S. When reagent A reaches the magnetic bead area of ​​the measuring cell 105, the quantum dots emit light; the electrochemical measuring cell 105 is not powered during the magnetic bead adsorption, washing, and luminescence process of the quantum dots. The luminescence signal and luminescence intensity of the quantum dots are detected by the multi-channel optical detection module 103; the luminescence wavelength of the quantum dots in this example is 785 nm, and a wavelength narrow band filter 107 is used in front of the optical detection module 103 for filtering; based on the luminescence intensity, the TSH signal intensity can be obtained.

[0207] After the quantum dots finish luminescence, reagent B with a magnetic bead cleaning effect is introduced into the flow channel 105a. Reagent B does not react with quantum dots and terpyridine ruthenium when no power is supplied. In the process of introducing reagent B into the flow channel 105a, the magnetic adsorption component 104 is close to the bottom of the measuring cell 105. Reagent B is filled at a speed of 200 μL / S. After the magnetic bead cleaning is completed, the measuring cell 105 is energized. In this case, 1.4V is used to excite electrochemical luminescence (terpyridine ruthenium and tripropylamine usually have an excitation voltage of 1.2V to 1.5V). A narrow-band filter 107 is installed at the front end of the photon detection and collection device 103. The luminescence is detected by the corresponding optical detection module 103, and the Tg signal intensity can be obtained.

[0208] After the luminescence is completed, similar to the common electrochemiluminescence measuring cell 105 , the electrochemiluminescence measuring cell 105 , the detection needle 102 , and the pipeline are cleaned by the reagent C, and a step-change voltage is applied during the cleaning process to ensure the cleaning effect.

[0209] Example 2

[0210] This embodiment is basically the same as Example 1, except that: the components of the TSH project and Tg project joint test kit include CdTe quantum dot-labeled TSH antibody (concentration of 2.0 μg / mL) and CuSe quantum dot-labeled Tg antibody (concentration of 0.5 μg / mL); wherein the specific components of the first reagent are: quantum dot-labeled TSH antibody 2.0 μg / mL, PBS 0.05M, Tween-200.5%wt, Triton X-100 0.5%wt, BSA3%wt, ProClin-300 0.2%wt, pH value is 7.4, and the specific components of the second reagent are: quantum dot-labeled Tg antibody Tg antibody 0.5 μg / mL, PBS 0.05M, Tween-200.5%wt, Triton X-100 0.5%wt, BSA3%wt, ProClin-300 0.2%wt, pH value is 7.4. CdTe quantum dot-labeled TSH antibody and CuSe quantum dot-labeled Tg antibody are both from Pumen Technology.

[0211] Example 3

[0212] The reagent composition of this embodiment is the same as that of embodiment 1, except that a two-step joint detection method is used, that is, the sample to be tested is placed in the reaction cup 106 with the first reagent and the second reagent respectively, mixed evenly, and then placed in the reaction module for full reaction, and then streptavidin-labeled magnetic beads are added, mixed evenly, and then placed in the reaction module for full reaction. After the reaction is completed, a chemiluminescent mixed sample and an electrochemiluminescent mixed sample are obtained respectively. The two mixed samples are sequentially passed into the flow channel 105a. For the specific process, please refer to Figure 3 As shown, the order of introduction is: introduce reagent B to activate the electrode by electrifying the electrode, introduce chemiluminescent mixed sample, introduce reagent D to clean the flow channel 105a and magnetic beads, introduce bubble isolation reagent D and reagent A, introduce reagent A for chemiluminescent detection, introduce reagent D to clean the flow channel 105a, introduce electrochemiluminescent mixed sample, introduce reagent B to clean the magnetic beads and perform electrochemiluminescent detection.

[0213] In this example, the emission wavelength of CdTe quantum dots is 785 nm, and the emission wavelength of CdSe quantum dots is 550 nm. A wavelength narrow-band filter 107 is used for filtering in front of the optical detection module 103; based on the emission intensity, the TSH and Tg signal intensities can be obtained respectively.

[0214] Example 4

[0215] This embodiment is basically the same as embodiment 3, except that the specific components of the second reagent are: 0.5 μg / mL of terpyridine ruthenium labeled Tg antibody, 0.1 M MES, 0.3% wt Tween-20, 0.5% wt Triton X-100, 2% wt BSA, 0.3% wt BND, and the pH value is 6.

[0216] Example 5

[0217] This embodiment is basically the same as embodiment 1, except that electrochemiluminescence is performed first and then chemiluminescence, and the order of reagent introduction is: introducing reagent B to energize the electrode to activate the electrode, introducing the sample mixture, introducing reagent D to clean the flow channel 105a and the magnetic beads, performing electrochemiluminescence detection with reagent B, introducing reagent D to clean the flow channel 105a and the magnetic beads, introducing bubble isolation reagent D and reagent A, and introducing reagent A for chemiluminescence detection.

[0218] Example 6

[0219] This embodiment is basically the same as embodiment 5, except that: the specific components of the second reagent are: CdSe quantum dot-labeled Tg antibody 0.5 μg / mL, PBS 0.05M, Tween-20 0.5%wt, Triton X-100 0.5%wt, BSA 3%wt, ProClin-300 0.2%wt, and the pH value is 7.4.

[0220] Example 7

[0221] This embodiment is basically the same as embodiment 5, except that a two-step joint detection method is used, that is, the sample to be tested is placed in a reaction cup 106 with the first reagent and the second reagent respectively, mixed evenly, and then placed in a reaction module for full reaction, and then streptavidin-labeled magnetic beads are added, mixed evenly, and then placed in the reaction module for full reaction. After the reaction is completed, a chemiluminescent mixed sample and an electrochemiluminescent mixed sample are obtained respectively, and the two mixed samples are sequentially passed into the flow channel 105a, first for electrochemiluminescence and then for chemiluminescence.

[0222] Example 8

[0223] This embodiment is basically the same as embodiment 7, except that: the specific components of the second reagent are: CdSe quantum dot-labeled Tg antibody 0.5 μg / mL, PBS 0.05M, Tween-20 0.5%wt, Triton X-100 0.5%wt, BSA 3%wt, ProClin-300 0.2%wt, and the pH value is 7.4.

[0224] Comparative Example 1

[0225] This comparative example is a comparative example of Examples 1 and 5. Tg is detected by electrochemiluminescence using the same equipment as in Example 1. The specific contents are basically similar to those in Example 1, except that the first reagent does not contain TSH antibody labeled with quantum dots, but contains TSH antibody labeled with acridinium ester. During the test, reagent A is not introduced into the flow channel 105a, but chemiluminescent co-reaction reagent H is introduced. 2 O 2 / NaOH solution.

[0226] Comparative Example 2

[0227] This comparative example is a comparative example of Examples 2 and 6. This comparative example is basically the same as comparative example 1, and the only difference is that the second reagent contains a Tg antibody labeled with quantum dots.

[0228] Comparative Example 3

[0229] This comparative example is a comparative example of Examples 3 and 7. This comparative example is substantially the same as comparative example 1, except that a two-step joint detection method is used.

[0230] Comparative Example 4

[0231] This comparative example is a comparative example of Examples 4 and 8. This comparative example is substantially the same as Comparative Example 2, except that the two-step joint detection method is used.

[0232] test:

[0233] Prepare 5 TSH / Tg serum samples of different concentrations. Use the detection methods of Examples 1 to 8 to detect the standard samples of different concentrations. Repeat the test three times for each sample to obtain test values ​​1-3 and calculate the average. The test results are as follows: Figure 1 And as shown in Table 1.

[0234] The master curve was established using the calibration samples with traceable values ​​in Examples 1 to 8 and Comparative Examples 1 to 4, and the two standard samples were tested. The results are shown in Table 2.

[0235] Table 1 Test sample results of different embodiments

[0236]

[0237]

[0238] Table 2 Test results of standard products in different embodiments and comparative examples

[0239]

[0240]

[0241]

[0242] It can be seen from Table 1 that different embodiments all have very high signal-to-noise ratios, indicating that both quantum dot chemiluminescence, quantum dot electrochemiluminescence, and terpyridine ruthenium electrochemiluminescence have very high detection capabilities.

[0243] From the comparison of the embodiments and comparative examples in Table 2, it can be seen that when using the one-step combined detection method, the use of quantum dot chemiluminescence-quantum dot electrochemiluminescence or terpyridine ruthenium electrochemiluminescence has higher accuracy than ordinary chemiluminescence. The reason may be that the emission wavelength of ordinary chemiluminescent substances is 420-470nm, which coincides with the absorption wavelength of quantum dots or terpyridine ruthenium, resulting in energy transfer during luminescence and affecting the accuracy of detection.

[0244] The TSH chemiluminescent signal-concentration relationship curve and the Tg electrochemiluminescent signal-concentration relationship curve of each embodiment are as follows: Figure 5 As shown in the figure, it can be seen that the higher the TSH and Tg concentrations, the higher the luminescence intensity.

[0245] In summary, the chemiluminescence-electrochemiluminescence co-detection equipment provided by the present invention can realize chemiluminescence and electrochemiluminescence in the same equipment. When there are multiple substances in the sample to be tested that need to be analyzed and detected, and both luminescence detection and electrochemiluminescence detection are required, the convenience of detection is greatly improved; and chemiluminescence detection in the flow channel can reduce individual differences in testing caused by sample containers compared to conventional chemiluminescence detection, and its accuracy is also higher.

[0246] The co-inspection device provided in a preferred embodiment of the present invention includes a control system, and the control system has a built-in timing control program. The timing control program realizes automatic sampling according to preset time nodes, thereby realizing automatic detection of chemiluminescence and electrochemiluminescence.

[0247] The chemiluminescence detection method provided by the present invention can realize electrochemiluminescence detection and chemiluminescence detection in the same device. When the sample requires both luminescence detection and electrochemiluminescence detection, the convenience of detection is greatly improved. Compared with conventional chemiluminescence detection, chemiluminescence detection can reduce individual differences in testing caused by sample containers and has higher accuracy.

[0248] The detection method provided in the preferred embodiment of the present invention is a combination of luminescent markers such as chemiluminescent markers (quantum dots) and electrochemiluminescent markers (terpyridine ruthenium), which can achieve time- and wavelength-resolved multiple detection through chemiluminescence and electrochemiluminescence;

[0249] The detection method provided in the preferred embodiment of the present invention uses quantum dots as chemiluminescent and electrochemiluminescent markers, which realize chemiluminescence and electrochemiluminescence at different time points by combining with chemiluminescent co-reactants and electrochemiluminescent co-reactants respectively.

[0250] When both the chemiluminescent marker and the electrochemiluminescent marker are quantum dots, the same quantum dots or different quantum dots can be selected. When the same quantum dots are selected, the same quantum dots can detect two analytes in one measuring cell through chemiluminescence and electrochemiluminescence; when different quantum dots are selected, multiple quantum dots can achieve multiple detections with time and wavelength resolution through chemiluminescence and electrochemiluminescence.

[0251] The above are only specific embodiments of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that can be easily thought of by a person skilled in the art within the technical scope disclosed by the present invention should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.

Claims

1. A chemical-electrochemical luminescence co-detection device, characterized in that: include: A reaction cup, a plurality of reagent containers, a detection needle, an injection control module, a measuring cell, an electrode assembly, a magnetic adsorption component and at least one set of optical detection modules; The reaction cup is used to hold a sample mixture, the sample mixture includes a conjugate, the conjugate is mainly obtained by specifically binding at least two analytes in the sample to be tested to at least two binding molecules in a one-to-one correspondence and adsorbing them onto magnetic beads, at least one of the at least two binding molecules is a molecule labeled with a chemiluminescent marker, and at least one of the at least two binding molecules is a molecule labeled with an electrochemiluminescent marker; At least a portion of the plurality of reagent holding containers are used to respectively hold a plurality of luminescent reagents, wherein the plurality of luminescent reagents include at least one chemiluminescent co-reactant and at least one electrochemiluminescent co-reactant; The detection needle is used to absorb the sample mixture from the reaction cup into the flow channel of the measuring cell, and is used to absorb the corresponding luminescent reagents from the multiple reagent containing containers into the flow channel of the measuring cell; The sample injection control module is used to control the detection needle to perform an aspiration action into the reaction cup and the multiple reagent holding containers; The measuring cell is used to provide a reaction space for realizing chemiluminescence and electrochemiluminescence; The electrode assembly is arranged in conjunction with the measuring cell, and the electrode assembly is used to excite the electrochemical luminescent marker to emit light after being energized; The magnetic adsorption element is arranged close to the measuring cell and is used to magnetically attract the magnetic beads in the flow channel of the measuring cell; The at least one set of optical detection modules is used to detect and collect the luminous intensity of the luminescent marker; The chemical-electrochemical luminescence co-detection device is configured as follows: The injection control module controls the detection needle to absorb the sample mixture into the flow channel of the measurement cell; After the conjugate is adsorbed by the magnetic adsorbent, the injection control module controls the detection needle to absorb the luminescent reagent into the flow channel of the measurement cell; When the absorbed luminescent reagent is the chemiluminescent co-reaction reagent, the chemiluminescent co-reaction reagent combines with the corresponding luminescent marker to achieve chemiluminescence, and the luminescence intensity is detected and collected by the at least one set of optical detection modules; when the absorbed luminescent reagent is the electrochemiluminescent co-reaction reagent, the electrode assembly is energized, the electrochemiluminescent co-reaction reagent combines with the corresponding luminescent marker to achieve electrochemiluminescence, and the luminescence intensity is detected and collected by the at least one set of optical detection modules.

2. The chemical-electrochemical luminescence co-detection device according to claim 1, characterized in that: The chemical-electrochemical luminescence co-detection device also includes a control system; The control system is in communication with the injection control module and the electrode assembly, and the control system has a built-in timing control program, which is configured to instruct the injection control module to control the detection needle to absorb the corresponding reagent from different reagent holding containers at different time nodes; When the current time node is the time node for absorbing the electrochemiluminescent co-reaction reagent, the control system sends an instruction to the electrode assembly to power it on. After the electrochemiluminescent reaction is completed, the control system controls the electrode assembly to power off.

3. The chemical-electrochemical luminescence co-detection device according to claim 2, characterized in that: The timing control program is configured to: control the sampling control module so that the detection needle first samples the chemiluminescent co-reaction reagent to achieve chemiluminescence, and then samples the electrochemiluminescent co-reaction reagent to achieve electrochemiluminescence; Alternatively, the timing control program is configured to: control the injection control module to make the detection needle first sample the electrochemiluminescent co-reaction reagent to achieve electrochemiluminescence, and then sample the chemiluminescent co-reaction reagent to achieve chemiluminescence.

4. The chemical-electrochemical luminescence co-detection device according to claim 2, characterized in that: A portion of the plurality of reagent holding containers is used to hold a neutral cleaning solution; and the timing control program is configured as follows: Controlling the sampling control module to make the detection needle sequentially perform sampling of the sample mixture, absorption of the neutral cleaning solution, sampling of the chemiluminescent co-reaction reagent, and sampling of the electrochemiluminescent co-reaction reagent; Alternatively, the sampling control module is controlled to enable the detection needle to sequentially perform sampling of the sample mixture, absorption of the neutral cleaning solution, sampling of the electrochemiluminescent co-reaction reagent, and sampling of the chemiluminescent co-reaction reagent.

5. The chemical-electrochemical luminescence co-detection device according to claim 4, characterized in that: The detection needle is also used to draw air into the flow channel of the measuring cell to form bubbles in the flow channel; the timing control program is configured as follows: Controlling the sampling control module to make the detection needle sequentially perform sampling of the sample mixture, aspiration of the neutral cleaning solution, aspiration of air, sampling of the chemiluminescent co-reaction reagent, and sampling of the electrochemiluminescent co-reaction reagent; Alternatively, the sampling control module is manipulated to cause the detection needle to sequentially perform sampling of the sample mixture, aspiration of the neutral cleaning solution, aspiration of air, sampling of the electrochemiluminescent co-reaction reagent, and sampling of the chemiluminescent co-reaction reagent.

6. The chemical-electrochemical luminescence co-detection device according to claim 1, characterized in that: A receiving groove is provided at the bottom of the outer wall of the measuring cell, and the receiving groove is used to receive at least part of the magnetic adsorption component; the magnetic adsorption component includes the following feature (1) or feature (2): (1) The magnetic adsorbent is a permanent magnet, and the chemiluminescence co-detection device further comprises a movement control module, which controls the magnetic adsorbent to move into the receiving groove to adsorb the magnetic beads at the beginning of the luminescence detection, and controls the magnetic adsorbent to move out of the receiving groove to release the magnetic adsorbent from adsorbing the magnetic beads after the luminescence detection. (2) The magnetic adsorption component is an electromagnet, and the electromagnet is configured to be powered on when the luminescence detection starts and powered off after the luminescence detection ends.

7. The chemical-electrochemical luminescence co-detection device according to claim 1, characterized in that: The number of the optical detection modules is 1 set, and the luminescent wavelengths of the luminescent markers used for detection can be detected and collected by the optical detection modules; Alternatively, there are multiple sets of optical detection modules, and multiple luminescent markers are used for detection. The multiple luminescent markers have different luminescent wavelengths, and the luminous intensity of the light emitted by the luminescent markers with different luminescent wavelengths is detected and collected by the corresponding optical detection modules respectively.

8. The chemical-electrochemical luminescence co-detection device according to claim 1, characterized in that: A transparent window is provided on the upper wall of the measuring cell, the at least one set of optical detection modules is provided above the transparent window, and a filter of a corresponding wavelength is provided between each set of the optical detection modules and the transparent window; Optionally, the filter is a narrow-band filter lens.

9. A chemi-electrochemiluminescence detection method, characterized in that: The steps include: A sample mixture is introduced into the flow channel of the measuring cell, wherein the sample mixture includes a conjugate, which is mainly obtained by fully reacting the sample to be tested, at least one first binding molecule, at least one second binding molecule and magnetic beads, and the conjugate is adsorbed on the bottom of the flow channel when passing near the magnetic adsorption member arranged outside the flow channel; A plurality of luminescent reagents are introduced into the flow channel of the measuring cell, each luminescent reagent is introduced at a different time point, and the plurality of luminescent reagents include at least one chemiluminescent co-reaction reagent and at least one electrochemiluminescent co-reaction reagent; Each of the first binding molecules is a molecule labeled with a chemiluminescent marker. When the number of the first binding molecules in the sample mixture is plural, the plural first binding molecules are molecules respectively labeled with different chemiluminescent markers, and each of the first binding molecules is a substance that can specifically bind to a corresponding analyte in the sample to be tested and adsorb to the magnetic beads; Each of the second binding molecules is a molecule labeled with an electrochemiluminescent marker. When the number of the second binding molecules in the sample mixture is multiple, the multiple second binding molecules are molecules labeled with different electrochemiluminescent markers, and each of the second binding molecules is a substance that can specifically bind to the corresponding analyte in the sample to be tested and adsorbed to the magnetic beads; The at least one chemiluminescent co-reaction reagent is bound to the luminescent marker on the at least one first binding molecule in a one-to-one correspondence to achieve chemiluminescence, and the corresponding luminescence intensity is detected and collected by the corresponding optical detection module; When the electrochemiluminescent co-reaction reagent is passed into the flow channel, the electrode assembly in the measuring cell is energized, and the at least one electrochemiluminescent co-reaction reagent is combined with the luminescent marker on the at least one second binding molecule in a one-to-one correspondence to realize electrochemiluminescence, and the corresponding luminescence intensity is detected and collected by the corresponding optical detection module.

10. The luminescence detection method according to claim 9, characterized in that: Before introducing different luminescent reagents into the flow channel of the measuring cell, the method further includes: introducing a neutral cleaning solution into the flow channel of the measuring cell.

11. The luminescence detection method according to claim 9, characterized in that: When the conjugate is adsorbed to the bottom of the flow channel by the magnetic adsorbent, a neutral cleaning solution is introduced into the flow channel to wash away free substances in the sample mixture that are not bound to the magnetic beads, and then the luminescent reaction reagent is introduced; Optionally, after the free substances not bound to the magnetic beads are washed away, before the luminescent reaction reagent is introduced, the process further includes introducing bubbles into the flow channel to isolate the neutral cleaning solution and the luminescent reaction reagent.

12. The luminescence detection method according to claim 11, characterized in that: The flow rate of the sample mixture in the flow channel is 10 μL / S to 30 μL / S, the flow rate of the chemiluminescent co-reaction reagent in the flow channel is 100 μL / S to 500 μL / S, and the flow rate of the electrochemiluminescent co-reaction reagent in the flow channel is 100 μL / S to 200 μL / S.

13. The luminescence detection method according to any one of claims 9 to 12, characterized in that: Also includes at least one of the following features (1) to (6); (1) After all luminescence detections are completed, an alkaline cleaning solution and the electrochemical luminescence co-reaction reagent are sequentially introduced into the flow channel, wherein the alkaline cleaning solution is a solution containing a detergent and a strong base, wherein the mass percentage of the detergent is 0.05% to 1%, the detergent is selected from at least one of Tween, Triton and polycatalpol, the strong base is selected from at least one of KOH and NaOH, and the pH of the alkaline cleaning solution is 12 to 14; (2) The electrochemiluminescent marker is terpyridine ruthenium or quantum dots; (3) The chemiluminescent marker is a quantum dot; (4) before using the device for electrochemiluminescence detection, including performing an electrode activation operation, during which an electrochemiluminescence co-reaction reagent is introduced into the flow channel; (5) The electrochemiluminescent co-reaction reagent contains a surfactant for washing the magnetic beads, wherein the surfactant is selected from at least one of Tween, Triton and polycatalpol, and the mass percentage of the surfactant is 0.05% to 1%; (6) The neutral cleaning solution is a solution containing phosphate and a detergent, the concentration of the phosphate is 10 mM to 500 mM, and the mass percentage of the detergent is 0.05% to 1%.

14. The luminescence detection method according to any one of claims 9 to 12, characterized in that: The electrochemiluminescent marker is terpyridine ruthenium, and the electrochemiluminescent co-reaction reagent corresponding to terpyridine ruthenium is a solution containing tripropylamine; Optionally, the solution containing tripropylamine is a phosphate solution of tripropylamine; Optionally, the concentration of tripropylamine in the tripropylamine phosphate solution is 0.1M to 0.2M, and / or the concentration of phosphate in the tripropylamine phosphate solution is 10mM to 500mM; Optionally, the concentration of terpyridine ruthenium in the tripropylamine phosphate solution is 0.01 nM to 5 μM.

15. The luminescence detection method according to any one of claims 9 to 12, characterized in that: The chemiluminescent marker is a quantum dot, and the chemiluminescent co-reaction reagent corresponding to the quantum dot is a solution containing potassium permanganate; Optionally, the solution containing potassium permanganate is a phosphate solution of potassium permanganate; Optionally, the concentration of potassium permanganate in the potassium permanganate phosphate solution is 1 mM to 3 mM, and / or the concentration of the phosphate is 1 mM to 100 mM; Optionally, the concentration of the quantum dots in the phosphate solution containing 0.5% Tween 20 by mass is 0.5 μM to 1.5 μM.

16. The luminescence detection method according to any one of claims 9 to 12, characterized in that: The first binding molecule is a quantum dot-labeled first detection object antibody, and the chemiluminescent co-reaction reagent is a solution containing potassium permanganate; The second binding molecule is a second antibody to be detected labeled with terpyridine ruthenium, and the electrochemiluminescent co-reaction reagent is a solution containing tripropylamine.

17. The luminescence detection method according to any one of claims 9 to 12, characterized in that: The chemiluminescent marker and the electrochemiluminescent marker are both quantum dots; The chemiluminescent co-reaction reagent is a solution containing potassium permanganate; the electrochemiluminescent co-reaction reagent is a solution containing tripropylamine; Optionally, the quantum dots are CdTe, CdSe, or CuInS2@ZnS.

18. The luminescence detection method according to any one of claims 9 to 12, characterized in that: The sample mixture is obtained by mixing the sample to be tested, the first reagent, the second reagent and the magnetic beads; The first reagent comprises 0.05 mg / L to 50.0 mg / L of a first antibody to be detected labeled with quantum dots, 1 mmol / L to 100 mmol / L of a buffered saline solution, 0.5 g / L to 50 g / L of a surfactant, and 0.05 g / L to 5 g / L of a preservative, and the pH value of the first reagent is 5.0 to 9.0; The second reagent comprises 0.05 mg / L to 50.0 mg / L of a second antibody to be detected labeled with terpyridine ruthenium, 1 mmol / L to 100 mmol / L of a buffered saline solution, 0.5 g / L to 50 g / L of a surfactant, and 0.05 g / L to 5 g / L of a preservative, and the pH value of the first reagent is 5.0 to 9.0; Alternatively, the second reagent includes 0.05 mg / L to 50.0 mg / L of quantum dot-labeled second antibody to be detected, 1 mmol / L to 100 mmol / L of buffered saline solution, 0.5 g / L to 50 g / L of surfactant, and 0.05 g / L to 5 g / L of preservative, and the pH value of the first reagent is 5.0 to 9.

0.

19. The luminescence detection method according to claim 18, characterized in that: The buffered saline solution includes one or more combinations of PBS, PB, PIPES, MES, HEPES, TRIS, HEPPS and Bis-Tris; The surfactant includes one or more of polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monooleate, polyethylene glycol tert-octylphenyl ether, polyethylene glycol nonylphenyl ether and fatty alcohol polyoxyethylene ether; The preservative includes one or more combinations of ProClin-300, ProClin-950, gentamicin and BND.

20. The luminescence detection method according to claim 16, characterized in that: The first object to be detected and the second object to be detected are respectively selected from PCT, TSH, HCG, CK-MB, MYO, NT-proBNP, Tg, CRP, PGI, PGII, cTnI, cTnT, CT, PTH, Ferritin, PIIINP, CIV, LN, PRL, FSH, LH, TBG, C-peptide, Insulin, H-FABP, Lp-PLA2, IL-6, HA, ProGRP, AMH, D-dimer, AFP, CEA, SAA, CA125, CA19-9, SCCA, NSE, CYFRA Any two of 21-1, CA15-3, CA72-4, HE4, PSA, HBsAg, HIVAg, IGFBP-3, Renin, ACTH, BNP, CA50, CA242, S100, MPO, hGH, SHBG, TNF-α, IL-1β, IL-2, IL-2R, IL-4, IL-5, IL-8, IL-10 and IL-17.

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