Chemical-electrochemical luminescence co-detection apparatus and method

Abstract

The present application relates to the technical field of light emission detection, and discloses a chemical-electrochemical light emission co-detection device and method. The chemical-electrochemical light emission co-detection device comprises a reaction cup, a plurality of reagent containers, a detection needle, a sample injection control module, a measuring pool, an electrode assembly, a magnetic attraction accessory and at least one set of optical detection module. The chemical-electrochemical light emission co-detection device is configured to: the detection needle sucks the sample mixture into the flow channel; after the combined substance is adsorbed by the magnetic attraction accessory, the detection needle sucks the light emission reagent into the flow channel; when the sucked light emission reagent is a chemical light emission co-reaction reagent, chemical light emission is realized; when the sucked light emission reagent is an electrochemical light emission co-reaction reagent, the electrode assembly is powered on to realize electrochemical light emission. The present application also provides a chemical light emission detection method, which can be implemented by the above device. The method can realize chemical light emission detection in an electrochemical light emission detection device, and has high accuracy.

Description

TECHNICAL FIELD

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

[0002] Electrochemiluminescence (ECL) utilizes electrochemiluminescence, which requires the application of a voltage to excite chemiluminescence, and has the advantages of high sensitivity, low background, long recycling and regeneration of luminescent substances, and accurate control of luminescence time. Electrochemiluminescence containers are usually performed in a closed flow channel to ensure consistent electrochemiluminescence conditions. A fixed luminescence container is called a measuring cell.

[0003] A chemiluminescence immunoassay analyzer is usually composed of one or more of a sample adding 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 obtained by calculation by the data processing system. In cooperation with the reagent, it is used for qualitative or quantitative analysis of the analyte in the human sample. Currently, the chemiluminescence detection method cannot be performed in an electrochemiluminescence detection device, and each test is generally a single test item. In addition, when there is an all-in-one machine for two chemiluminescence methods, or any chemiluminescence system is compatible, there is also no suitable detection method to enable an electrochemical and chemiluminescence all-in-one machine. The luminescence container of the all-in-one machine is in the reaction cup.

[0004] In view of this, the present application is proposed. SUMMARY

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

[0006] Embodiments of the present application can be implemented as follows:

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

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

[0009] The reaction cup is used to hold a sample mixture, and the sample mixture comprises a binding substance, which is obtained by specifically binding and adsorbing at least two detected substances in the sample to be detected to magnetic beads with at least two binding molecules, and at least one of the at least two binding molecules is a chemiluminescence label molecule, and at least one of the at least two binding molecules is an electrochemiluminescence label molecule;

[0010] At least one of the plurality of reagent containers is configured to hold a luminescent reagent, and the plurality of luminescent reagents includes at least one chemiluminescent co-reagent and at least one electrochemiluminescent co-reagent;

[0011] The detection needle is configured to draw the sample mixture from the reaction cup into the flow channel of the measurement cell and to draw the corresponding luminescent reagent from the plurality of reagent containers into the flow channel of the measurement cell;

[0012] The sample injection control module is configured to control the detection needle to perform the drawing action into the reaction cup and the plurality of reagent containers;

[0013] The measurement cell is configured to provide a reaction space for chemiluminescence and electrochemiluminescence;

[0014] The electrode assembly is configured to be coupled to the measurement cell and to excite the electrochemiluminescent label to emit light when energized;

[0015] The magnetic attraction accessory is configured to be disposed adjacent to the measurement cell and to magnetically attract the magnetic beads in the flow channel of the measurement cell;

[0016] The at least one optical detection module is configured to detect and collect the luminescent intensity of the luminescent label;

[0017] The chemiluminescence-electrochemiluminescence co-detection device is configured to:

[0018] The sample injection control module controls the detection needle to draw the sample mixture into the flow channel of the measurement cell;

[0019] After the binding substance is attracted by the magnetic attraction accessory, the sample injection control module controls the detection needle to draw the luminescent reagent into the flow channel of the measurement cell; when the drawn luminescent reagent is the chemiluminescent co-reagent, the chemiluminescent co-reagent combines with the corresponding luminescent label to realize chemiluminescence, and the luminescent intensity is detected and collected by the at least one optical detection module; when the drawn luminescent reagent is the electrochemiluminescent co-reagent, the electrode assembly is energized, the electrochemiluminescent co-reagent combines with the corresponding luminescent label to realize electrochemiluminescence, and the luminescent intensity is detected and collected by the at least one optical detection module.

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

[0021] The control system is communicatively connected with the sample injection control module and the electrode assembly, and the control system is built-in with a time sequence control program configured to instruct the sample injection control module to control the detection needle to draw the corresponding reagent from different reagent containers at different time nodes;

[0022] When the current time node is the time node of taking the electrochemiluminescence co-reagent, the control system issues an instruction to the electrode assembly to make it powered on. After the electrochemiluminescence reaction ends, the control system controls the electrode assembly to be powered off.

[0023] In an optional embodiment, the timing control program is configured to control the sample injection control module to make the detection needle first sample the chemiluminescence co-reagent to realize chemiluminescence, and then sample the electrochemiluminescence co-reagent to realize electrochemiluminescence.

[0024] Alternatively, the timing control program is configured to control the sample injection control module to make the detection needle first sample the electrochemiluminescence co-reagent to realize electrochemiluminescence, and then sample the chemiluminescence co-reagent to realize chemiluminescence.

[0025] In an optional embodiment, part of the plurality of reagent holding containers are used to hold neutral cleaning liquid; the timing control program is configured to:

[0026] control the sample injection control module to make the detection needle sequentially sample the sample mixture, take the neutral cleaning liquid, sample the chemiluminescence co-reagent, and sample the electrochemiluminescence co-reagent.

[0027] Alternatively, control the sample injection control module to make the detection needle sequentially sample the sample mixture, take the neutral cleaning liquid, sample the electrochemiluminescence co-reagent, and sample the chemiluminescence co-reagent.

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

[0029] control the sample injection control module to make the detection needle sequentially sample the sample mixture, take the neutral cleaning liquid, take air, sample the chemiluminescence co-reagent, and sample the electrochemiluminescence co-reagent.

[0030] Alternatively, control the sample injection control module to make the detection needle sequentially sample the sample mixture, take the neutral cleaning liquid, take air, sample the electrochemiluminescence co-reagent, and sample the chemiluminescence co-reagent.

[0031] In an optional embodiment, the bottom of the outer wall of the measuring cell is provided with a receiving groove for receiving at least part of a magnetic attraction member; the magnetic attraction member comprises the following features (1) or (2):

[0032] (1) The magnetic attraction member is a permanent magnet, and the chemiluminescence-electrochemiluminescence co-detection device further comprises a movement control module, which controls the magnetic attraction member to move into the receiving groove to adsorb the magnetic beads at the beginning of luminescence detection, and controls the magnetic attraction member to move out of the receiving groove to release the adsorption of the magnetic beads by the magnetic attraction member after the luminescence detection ends.

[0033] (2) The magnetic attraction member is an electromagnet, and the electromagnet is configured to be powered on at the start of the luminescence detection and powered off after the end of the luminescence detection.

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

[0035] Alternatively, the number of optical detection modules is multiple sets, and the luminescent markers used for detection are multiple, and the luminescent markers of different luminescence wavelengths have different luminescent intensities, and the luminescent intensities of the luminescent markers of different luminescence wavelengths are detected and collected by the corresponding optical detection modules.

[0036] In an optional embodiment, the upper wall of the measuring pool is provided with a transparent window, and at least one set of optical detection modules is arranged above the transparent window, and a corresponding wavelength filter is arranged 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 application provides a chemical-electrochemical luminescence detection method, comprising the following steps:

[0039] The sample mixture is introduced into the flow channel of the measuring pool, and the sample mixture comprises a conjugate, which is mainly obtained by fully reacting the sample to be measured, 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 attraction member arranged outside the flow channel;

[0040] A plurality of luminescent reagents are introduced into the flow channel of the measuring pool, and each luminescent reagent is introduced at a different time node, and the plurality of luminescent reagents comprise at least one chemical luminescence co-reagent and at least one electrochemical luminescence co-reagent;

[0041] Each first binding molecule is a molecule labeled with a chemical luminescence marker, when the number of first binding molecules in the sample mixture is multiple, the multiple first binding molecules are molecules labeled with different chemical luminescence markers respectively, and each first binding molecule is a substance that can specifically bind to a corresponding measured substance in the sample to be measured and be adsorbed on the magnetic beads;

[0042] Each second binding molecule is a molecule labeled with an electrochemical luminescence marker, when the number of second binding molecules in the sample mixture is multiple, the multiple second binding molecules are molecules labeled with different electrochemical luminescence markers respectively, and each second binding molecule is a substance that can specifically bind to a corresponding measured substance in the sample to be measured and be adsorbed on the magnetic beads;

[0043] The at least one chemiluminescence co-reagent is combined with the luminescent label on the at least one first binding molecule one by one to realize chemiluminescence, and the corresponding luminescent intensity is detected and collected by the corresponding optical detection module;

[0044] When the electrochemiluminescence co-reagent is introduced into the flow channel, the electrode assembly in the measuring cell is powered, the at least one electrochemiluminescence co-reagent is combined with the luminescent label on the at least one second binding molecule one by one to realize electrochemiluminescence, and the corresponding luminescent intensity is detected and collected by the corresponding optical detection module.

[0045] In an optional embodiment, before the different luminescent reagents are introduced into the flow channel of the measuring cell, a neutral cleaning liquid is introduced into the flow channel of the measuring cell.

[0046] In an optional embodiment, when the binding agent is adsorbed to the bottom of the flow channel by the magnetic adsorption member, the neutral cleaning liquid is introduced into the flow channel to wash away the free substance in the sample mixture that is not combined with the magnetic beads, and then the luminescent reaction reagent is introduced.

[0047] In an optional embodiment, after the free substance that is not combined with the magnetic beads is washed away, the luminescent reaction reagent is introduced before the gas bubbles are introduced into the flow channel to isolate the neutral cleaning liquid 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 chemiluminescence co-reagent in the flow channel is 100 μL / S to 500 μL / S, and the flow rate of the electrochemiluminescence co-reagent in the flow channel is 100 μL / S to 200 μL / S.

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

[0050] (1) After all luminescent detection is completed, the alkaline cleaning liquid and the electrochemiluminescence co-reagent are introduced into the flow channel in sequence, the alkaline cleaning liquid is a solution containing a detergent and a strong base, the mass percentage of the detergent is 0.05% to 1%, the detergent is selected from at least one of Tween, Triton, and polycarboxylic alcohol, the strong base is selected from at least one of KOH and NaOH, and the pH of the alkaline cleaning liquid is 12 to 14;

[0051] (2) The electrochemiluminescence label is terpyridine ruthenium or quantum dots;

[0052] (3) The chemiluminescence label is quantum dots;

[0053] (4) Before electrochemiluminescence detection is performed by using the device, an electrode activation operation is performed, and when the electrode activation operation is performed, the electrochemiluminescence co-reagent is introduced into the flow channel;

[0054] (5) The electrochemiluminescence co-reactant reagent contains a surfactant for cleaning the surface of the magnetic beads, the surfactant is selected from at least one of Tween, Triton and polyglycol, and the mass percentage of the surfactant is 0.05% to 1%;

[0055] (6) The neutral cleaning solution is a solution containing a 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%.

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

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

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

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

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

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

[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 the phosphate is 1 mM to 100 mM;

[0063] In an optional embodiment, the concentration of the quantum dot in the phosphate solution containing 0.5% Tween 20 by mass percentage 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 chemiluminescence co-reactant reagent is a solution containing potassium permanganate;

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

[0066] In an optional embodiment, both the chemiluminescence label and the electrochemiluminescence label are quantum dots;

[0067] The chemiluminescence co-reagent is a solution containing potassium permanganate; and the electrochemiluminescence co-reagent is a solution containing tripropylamine.

[0068] Optionally, the quantum dots are CdTe, CdSe, or CuInS2@ZnS.

[0069] In an optional embodiment, the first reagent comprises quantum dot-labeled first target antibody 0.05 mg / L-50.0 mg / L, buffer salt solution 1 mmol / L-100 mmol / L, surfactant 0.5 g / L-50 g / L, and preservative 0.05 g / L-5 g / L, and the pH value of the first reagent is 5.0-9.0.

[0070] The second reagent comprises terpyridine ruthenium-labeled second target antibody 0.05 mg / L-50.0 mg / L, buffer salt solution 1 mmol / L-100 mmol / L, surfactant 0.5 g / L-50 g / L, and preservative 0.05 g / L-5 g / L, and the pH value of the first reagent is 5.0-9.0.

[0071] Alternatively, the second reagent comprises quantum dot-labeled second target antibody 0.05 mg / L-50.0 mg / L, buffer salt solution 1 mmol / L-100 mmol / L, surfactant 0.5 g / L-50 g / L, and preservative 0.05 g / L-5 g / L, and the pH value of the first reagent is 5.0-9.0. The buffer salt solution comprises a combination of one or more of PBS (phosphate buffered saline), PB (phosphate), PIPES (piperazine-1, 4-bisethanesulfonic acid), MES (2-(N-morpholino) ethanesulfonic acid), HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid), TRIS (tris-hydroxymethyl aminomethane), HEPPS (4-(2-hydroxyethyl)-1-piperazine propyl sulfonic acid), and Bis-Tris (bis (2-hydroxyethyl) amino-tris (hydroxymethyl) methane).

[0072] In an optional embodiment, the surfactant comprises a combination of one or more of polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monooleate, polyethylene glycol tert-octyl phenyl ether, polyethylene glycol nonyl phenyl ether, and fatty alcohol polyoxyethylene ether.

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

[0074] In optional embodiments, the first and second analytes are selected from any two of 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 (cardiac troponin I), cTnT (cardiac 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 (thyroxine binding globulin), C-peptide (C-peptide), Insulin (insulin), H-FABP (heart-type fatty acid binding protein), Lp-PLA2 (lipoprotein-associated phospholipase A2), IL-6 (interleukin 6), HA (hyaluronic acid), ProGRP (pro-gastrin-releasing peptide), AMH (anti-mullerian hormone), D-dimer (D-Dimer), AFP (alpha-fetoprotein), CEA (carcinoembryonic antigen), SAA (serum amyloid A), CA125 (cancer antigen 125), CA19-9 (cancer 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 (cancer antigen 15-3), CA72-4 (cancer 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 (cancer antigen 50), CA242 (cancer antigen 242), S100 (S100 protein), MPO (myeloperoxidase), hGH (human growth hormone), SHBG (sex hormone binding globulin), TNF-a (tumor necrosis factor-a), IL-1b (interleukin-1b), 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] Advantages of embodiments of the application include, for example:

[0076] The chemical-electrochemical luminescence co-detection device provided by the application can realize chemical luminescence and electrochemical luminescence in the same device through the cooperation of a reaction cup, a plurality of reagent containing containers, a detection needle, a sample injection control module, a measuring pool, an electrode assembly, a magnetic attraction accessory and at least one set of optical detection modules.

[0077] The chemical-electrochemical luminescence detection method provided by the application can realize chemical luminescence and electrochemical luminescence detection at different time nodes in the measuring pool of the same electrochemical detection device, and has high convenience and high accuracy. BRIEF DESCRIPTION OF DRAWINGS

[0078] In order to more clearly illustrate the technical solutions of the embodiments of the application, the drawings needed in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some of the embodiments of the application, and therefore should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can also be obtained without creative labor on the basis of these drawings.

[0079] Figure 1 The device structure diagram of the method provided by the application is provided.

[0080] Figure 2 The flowchart of the reagent of the first kind provided by the application into the flow channel in one-step joint detection is provided.

[0081] Figure 3 The flowchart of the reagent of the first kind provided by the application into the flow channel in two-step joint detection is provided.

[0082] Figure 4 The timing control diagram of the embodiments of the application is provided.

[0083] Figure 5 The TSH chemical luminescence signal-concentration relationship curve and the Tg electrochemical luminescence signal-concentration relationship curve of each embodiment are provided.

[0084] Figure: 101-reagent containing container; 102-detection needle; 103-optical detection module; 104-magnetic attraction accessory; 105-measuring pool; 105a-flow channel; 106-reaction cup; 107-filter; 108-receiving groove; A-chemical luminescence co-reaction reagent; B-electrochemical luminescence co-reaction reagent; C-alkaline cleaning liquid; D-neutral cleaning liquid; S-sample mixture. DETAILED DESCRIPTION

[0085] In order to make the objects, technical solutions and advantages of the embodiments of the present application clearer, the following will clearly and completely describe the technical solutions in the embodiments of the present application with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only some but not all of the embodiments of the present application. The components of the embodiments of the present application described and shown in the drawings can be arranged and designed in various different configurations.

[0086] Therefore, the following detailed description of the embodiments of the present application provided in the drawings is not intended to limit the scope of the claimed present application, but only represents selected embodiments of the present application. All other embodiments obtained by those of ordinary skill in the art based on the embodiments in the present application without creative labor are within the scope of protection of the present application.

[0087] It should be noted that: similar reference numerals and letters represent similar items in the following drawings, therefore, once an item is defined in one drawing, it does not need to be further defined and explained in the subsequent drawings.

[0088] In the description of the present application, it should be noted that if the terms "upper", "lower", "inner", "outer" and the like indicate the orientation or positional relationship based on the orientation or positional relationship shown in the drawings, or the orientation or positional relationship when the product of the present application is usually placed, which is only for the convenience of describing the present application and simplifying the description, and does not indicate or imply that the indicated device or element must have a particular orientation, be constructed and operated in a particular orientation, therefore, it cannot be understood as a limitation on the present application.

[0089] In addition, if the terms "first", "second" and the like appear, they are only used to distinguish the description, and cannot be understood as indicating or implying relative importance.

[0090] It should be noted that the features in the embodiments of the present application can be combined with each other without conflict.

[0091] Please refer to Figure 1 , Figure 1 The electrochemical luminescence co-detection device provided by the embodiments of the present application comprises:

[0092] The reaction cup 106, the plurality of reagent containers 101, the detection needle 102, the sample injection control module, the measuring cell 105, the electrode assembly, the magnetic attraction accessory 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 combination; the combination is obtained by specific binding of at least two analytes in a sample to be tested to at least two binding molecules one by one and adsorption to magnetic beads; at least one of the at least two binding molecules is a chemiluminescent label labeled molecule, and at least one of the at least two binding molecules is an electrochemiluminescent label labeled molecule;

[0094] At least a part of the plurality of reagent holding containers 101 is used to hold a plurality of luminescent reagents respectively, and the plurality of luminescent reagents includes at least one chemiluminescent co-reagent and at least one electrochemiluminescent co-reagent;

[0095] The detection needle 102 is used to suck the sample mixture in the reaction cup 106 into the flow channel 105a of the measuring pool 105, and is used to suck the corresponding luminescent reagent in the plurality of reagent holding containers 101 into the flow channel 105a of the measuring pool 105;

[0096] The sample injection control module is used to control the detection needle 102 to perform the sucking action into the reaction cup 106 and the plurality of reagent holding containers 101;

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

[0098] The electrode assembly is matched with the measuring pool 105, and is used to excite the electrochemiluminescent label to emit light after being powered on;

[0099] The magnetic attraction accessory 104 is arranged close to the measuring pool 105, and is used to realize magnetic attraction to the magnetic beads in the flow channel 105a of the measuring pool 105;

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

[0101] The chemiluminescence-electrochemiluminescence co-detection device is configured to:

[0102] The sample injection control module controls the detection needle 102 to suck the sample mixture into the flow channel 105a of the measuring pool 105;

[0103] After the combination to be combined is adsorbed by the magnetic attraction accessory 104, the sample injection control module controls the detection needle 102 to suck the luminescent reagent into the flow channel 105a of the measuring pool 105, when the sucked luminescent reagent is a chemiluminescent co-reagent, the chemiluminescent co-reagent is combined with the corresponding luminescent label to realize chemiluminescence, and the luminescent intensity is detected and collected by at least one set of optical detection module 103;

[0104] When the extracted luminescent reagent is an electrochemiluminescence co-reagent, the electrode assembly is powered on, the electrochemiluminescence co-reagent is combined with the corresponding luminescent label to realize electrochemiluminescence, and the luminescent intensity is detected and collected by at least one set of optical detection module 103.

[0105] The chemical-electrochemical luminescence co-detection device provided by the application is suitable for chemical luminescence and electrochemiluminescence detection of a sample to be detected with multiple analytes. After the binding molecules labeled with luminescent labels are mixed and reacted with the sample to be detected and the magnetic beads, only one sample injection is needed, and subsequent introduction of corresponding chemical luminescence co-reagents and electrochemiluminescence co-reagents at different time nodes can realize chemical luminescence detection and electrochemiluminescence detection, which is simple to operate and has high luminescence accuracy in the flow channel 105a.

[0106] It should be noted that the co-detection device provided by the application can have one or multiple chemical luminescence detection times in the whole luminescence detection process. To realize multiple chemical luminescence detections, multiple chemical luminescence labels are needed to label different molecules combined with magnetic beads, and correspondingly, the types of chemical luminescence co-reagents introduced into the flow channel 105a are also multiple and matched with the chemical luminescence labels.

[0107] Similarly, the electrochemiluminescence detection can have one or multiple times in the whole luminescence detection process. To realize multiple electrochemiluminescence detections, multiple electrochemiluminescence labels are needed to label different molecules combined with magnetic beads, and correspondingly, the types of electrochemiluminescence co-reagents introduced into the flow channel 105a are also multiple and matched with the electrochemiluminescence labels.

[0108] The chemical luminescence detection and the electrochemiluminescence detection share the measuring pool 105 as a luminescence container. The measuring pool 105 can trigger electrochemiluminescence reaction or chemical luminescence reaction. Further, the structure of the measuring pool 105 is basically the same as that of the conventional electrochemiluminescence detection measuring pool 105. Specifically, the electrode assembly is arranged in cooperation with the measuring pool 105, and the electrode assembly has 2 or 3 electrodes. When the electrode assembly has 3 electrodes, they are working electrode (WE), auxiliary electrode (CE) and reference electrode (RE) respectively. When the electrode assembly has 2 electrodes, it does not include the reference electrode (RE). Among them, platinum electrode is used as working electrode (WE) and auxiliary electrode (CE), and silver chloride electrode is used as reference electrode (RE). Generally, the working electrode (WE) is located in the flow channel 105a of the measuring pool 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 the transparent window of the luminescence area.

[0109] The optical detection module 103 further comprises a magnetic attraction member 104. The magnetic attraction member 104 is located at the bottom of the measuring pool 105. The magnetic attraction member 104 is capable of moving towards the bottom of the measuring pool 105 and attracting the magnetic micro-particles in the measuring pool 105. The magnetic attraction member 104 is capable of moving away from the bottom of the measuring pool 105 to release the attraction to the magnetic micro-particles in the measuring pool 105. Further, the magnetic attraction member 104 is capable of moving by being displaced up and down relative to the measuring pool 105, or by being rotated relative to the measuring pool 105.

[0110] Specifically, the bottom of the outer wall of the measuring pool 105 is provided with a receiving groove 108 for receiving at least part of the magnetic attraction member 104. The magnetic attraction member 104 is capable of moving into the receiving groove 108 towards the bottom of the measuring pool 105 to attract the magnetic micro-particles in the measuring pool 105. The provision of the receiving groove 108 facilitates the loading and removal of the magnetic attraction member 104. Specifically, the receiving groove 108 is formed by the bottom of the measuring pool 105 being inwardly recessed. The receiving groove 108 is located substantially opposite to the electrode sheet of the light-emitting region, which helps the magnetic micro-particles in the measuring pool 105 to be attracted by the magnetic attraction member 104 and stay on the surface of the electrode sheet.

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

[0112] Optionally, when the magnetic attraction member 104 is a permanent magnet, the chemical-electrochemical luminescence co-detection device further comprises a movement control module. The movement control module controls the magnetic attraction member 104 to move into the receiving groove 108 at the beginning of the luminescence detection, and controls the magnetic attraction member 104 to move out of the receiving groove 108 after the luminescence detection is completed to release the attraction of the magnetic attraction member 104 to the magnetic beads.

[0113] Optionally, when the magnetic attraction member 104 is an electromagnet. The electromagnet loses magnetic force after being powered off, so there is no need to additionally provide a movement mechanism for controlling the movement of the electromagnet.

[0114] Further, the number of optical detection modules 103 is one or more, which are installed above the light-emitting region transparent window of the measuring pool 105. The sensor of each set of optical detection modules 103 is provided with a filter 107 of a specific wavelength on the window. The filter 107 is used to distinguish the luminescence intensity of different luminescent objects. Specifically, the band-pass filter wavelength can be selected by switching the filter 107. The filter 107 is a narrow-band filter lens. According to the wavelength of the luminescent object, the corresponding narrow-band filter lens is matched to improve the anti-interference performance of each co-detection item and avoid mutual influence between the co-detection items.

[0115] It should be noted that the number of optical detection modules 103 is not limited to multiple sets. If the use of multiple luminescent substances meets the measurement range of each item with the degree of distinction of the co-reaction reagent, the number of optical detection modules 103 can also be one set. Setting one set or multiple sets of optical detection modules 103 depends on whether the luminescent wavelength can be received by the photomultiplier tube with high sensitivity.

[0116] Optionally, each set of optical detection module 103 includes a photon detector and a collector; the photon detector is an avalanche photodiode (APD), a silicon photomultiplier (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 I-V conversion circuit, or a photon counter.

[0117] Optionally, the chemical-electrochemical luminescence co-detection device can further include a reaction module, which mainly provides a better reaction environment to promote the reaction or binding of the sample to be measured, the binding molecules and the magnetic beads. Specifically, the reaction module can be a heater.

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

[0119] The control system is in communication connection with the sample injection control module and the electrode assembly. The control system is built-in with a time sequence control program, which is configured to instruct the sample injection control module to control the detection needle to suck the corresponding reagent from the different reagent containers 101 at different time nodes.

[0120] When the current time node is the time node for sucking the electrochemical luminescence co-reaction reagent, the control system issues an instruction to the electrode assembly to make it powered on. After the electrochemical luminescence reaction is completed, the control system controls the electrode assembly to be powered off.

[0121] The setting of the control system and the time sequence control program can realize the automatic sampling of the chemical luminescence and the electrochemical luminescence at a specific time node, and control the electrode assembly to be powered on at a suitable time node, thereby realizing the automatic detection of the chemical luminescence and the electrochemical luminescence.

[0122] In addition, the control system is also in communication connection with the optical detection module 103. 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 measured substance.

[0123] Optionally, the time sequence control program is configured to control the sample injection control module to make the detection needle 102 first sample the chemical luminescence co-reaction reagent to realize the chemical luminescence, and then sample the electrochemical luminescence co-reaction reagent to realize the electrochemical luminescence.

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

[0125] It should be noted that when the chemiluminescence detection is multiple times and the electrochemiluminescence detection is multiple times in the whole detection process, for example, the timing control program can be configured to sequentially perform chemiluminescence co-reagent sampling, electrochemiluminescence co-reagent sampling, chemiluminescence co-reagent sampling, and electrochemiluminescence co-reagent sampling; or electrochemiluminescence co-reagent sampling, electrochemiluminescence co-reagent sampling, chemiluminescence co-reagent sampling, and chemiluminescence co-reagent sampling.

[0126] Optionally, part of the plurality of reagent containers 101 are used to contain neutral cleaning liquid;

[0127] The timing control program is configured to control the sample injection control module to make the detection needle 102 sequentially sample the sample mixture, suck the neutral cleaning liquid, sample the chemiluminescence co-reagent, and sample the electrochemiluminescence co-reagent.

[0128] Alternatively, the timing control program is configured to control the sample injection control module to make the detection needle 102 sequentially sample the sample mixture, suck the neutral cleaning liquid, sample the electrochemiluminescence co-reagent, and sample the chemiluminescence co-reagent.

[0129] That is, the timing control program is configured to make the detection needle 102 suck the neutral cleaning liquid after the sample mixture sampling is completed, and suck the neutral cleaning liquid between the sampling of the two luminescence reagents.

[0130] Optionally, the detection needle 102 is also used to suck air into the flow channel 105a of the measuring pool 105 to form a bubble in the flow channel 105a.

[0131] The timing control program is configured to control the sample injection control module to make the detection needle 102 sequentially sample the sample mixture, suck the neutral cleaning liquid, suck the air, sample the chemiluminescence co-reagent, and sample the electrochemiluminescence co-reagent.

[0132] The timing control program is configured to control the sample injection control module to make the detection needle 102 sequentially sample the sample mixture, suck the neutral cleaning liquid, suck the air, sample the electrochemiluminescence co-reagent, and sample the chemiluminescence co-reagent.

[0133] That is, the timing control program is configured to make the detection needle 102 suck the air after the neutral cleaning liquid sucking is completed, to isolate the neutral cleaning liquid and the subsequent luminescence reagent.

[0134] The chemical-electrochemical luminescence detection method provided by the embodiment of the present application comprises the following steps:

[0135] The sample mixture is introduced into the flow channel 105a of the measuring pool 105, and the sample mixture comprises a complex obtained by fully reacting a sample to be measured, at least one first binding molecule, at least one second binding molecule and magnetic beads, and the complex is adsorbed on the bottom of the flow channel 105a when passing near the magnetic adsorption member 104 arranged outside the flow channel 105a;

[0136] The plurality of luminescent reagents are introduced into the flow channel 105a of the measuring pool 105, and each luminescent reagent is introduced at a different time node, and the plurality of luminescent reagents comprise at least one chemical luminescence co-reagent and at least one electrochemical luminescence co-reagent;

[0137] Each first binding molecule is a molecule labeled by a chemical luminescence label, when the number of first binding molecules in the sample mixture is multiple, the multiple first binding molecules are molecules labeled by different chemical luminescence labels respectively, and each first binding molecule is a substance capable of specifically binding to a corresponding measured substance in the sample to be measured and adsorbed on the magnetic beads;

[0138] Each second binding molecule is a molecule labeled by an electrochemical luminescence label, when the number of second binding molecules in the sample mixture is multiple, the multiple second binding molecules are molecules labeled by different electrochemical luminescence labels respectively, and each second binding molecule is a substance capable of specifically binding to a corresponding measured substance in the sample to be measured and adsorbed on the magnetic beads;

[0139] At least one chemical luminescence co-reagent is combined with the luminescent label on at least one first binding molecule one by one to realize chemical luminescence, and the corresponding luminescent intensity is detected and collected by the optical detection module 103 of the device;

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

[0141] The detection method provided by the application can realize electrochemiluminescence detection and chemiluminescence detection in the same device, thereby greatly improving the convenience of detection when the sample needs to be detected by both chemiluminescence and electrochemiluminescence; compared with conventional chemiluminescence detection, the chemiluminescence detection in the flow channel 105a can reduce the individual differences caused by the sample container, and the accuracy is higher. The detection method provided by the application can be implemented by the device provided by the embodiment of the application, or can be implemented by an existing electrochemiluminescence detection device.

[0142] It should be noted that the detection method provided by the application can detect multiple substances in the sample to be detected, and can perform at least one chemiluminescence detection and at least one electrochemiluminescence detection. Several chemiluminescence detections correspond to several chemiluminescence markers, several first binding molecules, and several times of adding chemiluminescence co-reaction reagents (the multiple times of adding chemiluminescence co-reaction reagents can be the same or different, and the principle of adding is that the chemiluminescence co-reaction reagent can combine with the chemiluminescence marker to emit light); several electrochemiluminescence detections correspond to several electrochemiluminescence markers, several second binding molecules, and several times of adding electrochemiluminescence co-reaction reagents (the multiple times of adding electrochemiluminescence co-reaction reagents can be the same or different, and the principle of adding is that the electrochemiluminescence co-reaction reagent can combine with the electrochemiluminescence marker to emit light).

[0143] Specifically, taking one chemiluminescence detection and one electrochemiluminescence detection as an example, the specific way of chemiluminescence detection and electrochemiluminescence detection in the same electrochemiluminescence detection device is, for example:

[0144] I. The first kind (one-step joint detection: sample and detection reagent are added at one time):

[0145] First, the chemiluminescence detection is performed, and then the electrochemiluminescence detection is performed, and the detection method is specifically:

[0146] S1, sample mixing reaction

[0147] The sample to be detected, at least one first reagent, and at least one second reagent are placed in the reaction cup 106 and mixed uniformly, and then placed in the reaction module for sufficient reaction, and then streptavidin-coated magnetic beads are added, mixed, and then placed in the reaction module for sufficient reaction. The magnetic beads have a particle size of 1 μm to 3.0 μm, and the balance of the magnetic bead loss rate and the cleaning effect of the measuring pool 105 is achieved. After the reaction is completed, a sample mixture is obtained.

[0148] The first reagent is a first binding molecule labeled with a chemiluminescent label, and the first binding molecule is a substance capable of specifically binding to a first analyte in the sample to be measured; the second reagent is a second binding molecule labeled with an electrochemiluminescent label, and the second binding molecule is a substance capable of specifically binding to a second analyte in the sample to be measured and adsorbing to the magnetic beads.

[0149] In this step, the sample to be measured, the first reagent, the second reagent and the magnetic beads are fully reacted to make the first binding molecule labeled with the chemiluminescent label and the first analyte bind to the magnetic beads, and to make the second binding molecule labeled with the electrochemiluminescent label and the second analyte bind to the magnetic beads to generate a conjugate.

[0150] Optionally, the chemiluminescent label is a quantum dot, and the chemiluminescent co-reactant corresponding to the quantum dot (reagent A) is a solution containing potassium permanganate. Further, the solution containing potassium permanganate is a potassium permanganate phosphate solution. 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. The concentration of the quantum dot in the phosphate solution containing 0.5% (mass percentage) Tween 20 is 0.5 μM to 1.5 μM, which can ensure that the luminescence intensity of the quantum dot is comparable to the electrochemiluminescence.

[0151] Optionally, the electrochemiluminescent label is tris (2-phenylpyridine) ruthenium, and the electrochemiluminescent co-reactant corresponding to the tris (2-phenylpyridine) ruthenium (reagent B) is a solution containing tripropylamine. Further, the solution containing tripropylamine is a tripropylamine phosphate solution. The concentration of tripropylamine in the tripropylamine phosphate solution is 0.1 M to 0.2 M, and / or the concentration of phosphate in the tripropylamine phosphate solution is 10 mM to 500 mM. To ensure the luminescence intensity, the concentration of the tris (2-phenylpyridine) ruthenium in the tripropylamine phosphate solution is 0.01 nM to 5 μM.

[0152] Particularly preferably, both the chemiluminescent label and the electrochemiluminescent label are quantum dots; optionally, the quantum dots are CdTe, CdSe or CuInS2@ZnS. It should be noted that when both the chemiluminescent label and the electrochemiluminescent label are quantum dots, the same kind of quantum dots can be selected, or different kinds of quantum dots can be selected. When the same kind of quantum dots are selected, the same quantum dots can be used to detect two analytes in one measuring cell 105 through chemiluminescence and electrochemiluminescence; when different kinds of quantum dots are selected, multiple quantum dots can be used to achieve time and wavelength-resolved multiplex detection 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 comprises 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) 0.01-0.1 M (e.g. 0.01 M, 0.02 M, 0.05 M or 0.1 M), 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 (bovine serum albumin) 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 (e.g. 0.05% wt, 0.1% wt, 0.2% wt or 0.5% wt), and the pH of the first reagent is 7.3-7.5 (e.g. 7.3, 7.4 or 7.5).

[0155] The PBS in the component provides a buffer environment, and the above concentration range ensures pH stability. If the concentration is too high, the protein will be denatured. The Tween-20 and Triton X-100 in the component protect the high-order structure of the protein, and the above concentration range ensures the activity and stability of the antibody. If the concentration is too high, the protein will be inactivated. The BSA in the component blocks the non-specific binding sites in the reaction, and the above concentration range reduces the interference of non-specific adsorption. If the concentration is too high, the luminescence signal will be reduced. The ProClin-300 in the component acts as a bacteriostatic preservative, and the above concentration range ensures the long-term stability of the solution. If the concentration is too high, the luminescence signal will be reduced. The pH range in the first reagent is the optimal buffer range of PBS.

[0156] Optionally, the second reagent comprises 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) 0.02-0.2 M (e.g. 0.02 M, 0.05 M, 0.1 M or 0.2 M), 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 (bovine serum albumin) 0.5-5% wt (e.g. 0.5% wt, 0.8% wt, 1% wt, 3% wt or 5% wt), BND (5-bromo-5-nitro-l,3-dioxane) 0.05-0.5% wt (e.g. 0.05% wt, 0.1% wt, 0.2% wt or 0.5% wt), and the pH of the second reagent is 5.9-6.1 (e.g. 5.9, 6 or 6.1).

[0157] The MES in the component provides a buffer environment, and the pH is stable in the above concentration range. If the concentration is too high, the protein will be denatured. The Tween-20 and Triton X-100 in the component protect the high-order structure of the protein, and the antibody remains active and stable in the above concentration range. If the concentration is too high, the protein will be inactivated. The BSA in the component blocks the non-specific binding sites in the reaction, and the non-specific adsorption interference is reduced in the above concentration range. If the concentration is too high, the luminescence signal will be reduced. The BND in the component acts as a bacteriostatic preservative, and the solution is stable for a long time in the above concentration range. If the concentration is too high, the luminescence signal will be reduced. 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 0.01-0.1 M (e.g., 0.01 M, 0.02 M, 0.05 M, or 0.1 M), 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 (e.g., 0.05% wt, 0.1% wt, 0.2% wt, or 0.5% wt), and the pH of the second reagent is 7.3-7.5 (e.g., 7.3, 7.4, or 7.5).

[0159] S2, the reagent is sequentially introduced into the flow channel 105a

[0160] The specific flow is shown in Figure 2 After starting the electrochemiluminescence detection device, the detection needle 102 is used to suck the electrochemiluminescence co-reaction reagent (reagent B) into the flow channel 105a under time sequence control, and the electrode is powered on to activate the electrode.

[0161] Then the detection needle 102 sucks the sample mixture (S) and the sample mixture is left in the measuring pool 105. The magnet below the measuring pool 105 moves and tightly fits into the containing groove 108 below the light-emitting area at the bottom of the measuring pool 105. The complex slowly flows through the reaction flow channel 105a of the measuring pool 105 under time sequence control, and the magnetic beads in the reaction complex are adsorbed to the top of the magnetic adsorption member 104. The flow rate is usually 10-30 μL / S, and the magnetic separation position of the measuring pool 105 is flat to ensure the effective magnetic bead adsorption area and the cleaning effect.

[0162] Then the detection needle 102 sucks the neutral cleaning liquid (reagent D) into the flow channel 105a for repeated cleaning of the magnetic beads, and the unbound magnetic beads are carried away by the reagent D under the action of the fluid.

[0163] Then the detection needle 102 sucks enough bubbles to cover the flow channel 105a into the flow channel 105a, which is used to isolate the reagent D and the reagent A, for example, 10-50 μL of bubbles are used for isolation.

[0164] Then the reagent A is sucked into the flow channel 105a by the detection needle 102, and flows through the measuring cell 105 at a flow rate of 100 μL / S to 500 μL / S, so as to ensure that the reagent A is rapidly combined with the magnetic beads combined with the quantum dots, is not diluted by the residual reagent D, and the magnetic beads are not washed away. When the reagent A reaches the magnetic bead area of the measuring cell 105, the quantum dots emit light; the quantum dot adsorption process, the magnetic bead cleaning process, and the light emission process are not powered on the electrochemical measuring cell 105. The light emission of the quantum dots is detected by the photon detection and collection device 103; generally, there are condenser lenses and spectrometer lenses between the photon detection and collection device 103 and the measuring cell 105.

[0165] After the quantum dots complete light emission, the electrochemiluminescence co-reagent (reagent B) with a magnetic bead cleaning effect is sucked into the flow channel 105a by the detection needle 102, and the reagent B does not react with the quantum dots and the trispyridine ruthenium without being powered on. During the process of the reagent B flowing into the flow channel 105a, the magnetic adsorption member 104 is attached to the bottom of the measuring cell 105, and the adsorption of the magnetic beads is maintained. The reagent filling is completed at a speed of 100 μL / S to 200 μL / S, and after the magnetic bead cleaning is completed, the measuring cell 105 is powered on. If the electrochemiluminescence label is trispyridine ruthenium, 1.4 V excitation electrochemiluminescence (the excitation voltage of trispyridine ruthenium and tripropylamine is generally 1.2 V to 1.5 V) can be used. After the trispyridine ruthenium emits light, it is detected by the photon detection and collection device 103.

[0166] Optionally, in this step, a neutral reagent (reagent D) can also be flowed in before the reagent B is flowed in.

[0167] After the light emission is completed, the reagent C is flowed into the flow channel 105a, so as to clean the measuring cell 105, the detection needle 102, and the pipeline by the reagent C. A step change voltage is applied during the cleaning process to ensure the cleaning effect. The applied voltage is different according to the electrode material of the electrochemical measuring cell 105. After the cleaning is completed, the electrode surface is reduced by being powered on, and the measuring cell 105 is re-pre-filled.

[0168] Optionally, the electrochemiluminescence co-reagent (reagent B) contains a surfactant for cleaning the magnetic beads. The surfactant is selected from at least one of Tween, Triton, and polycarboxylic alcohol, 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. The mass percentage of the detergent is 0.05% to 1%, the detergent is selected from at least one of Tween, Triton, and polycarboxylic alcohol, 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 a phosphate and a detergent, the concentration of the phosphate is 10 mM-500 mM, and the mass percentage of the detergent is 0.05%-1%.

[0171] II. The second method (one-step combined detection: the sample and the detection reagent are put into once):

[0172] First, electrochemiluminescence detection is performed, and then chemiluminescence detection is performed, and the detection mode is specifically as follows:

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

[0174] S1, sample mixing reaction

[0175] The sample to be detected, the first reagent and the second reagent are mixed uniformly in the reaction cup 106, and then the streptavidin magnetic beads are added, mixed, and then put into the reaction module for full reaction. After the reaction is completed, a sample mixture is obtained.

[0176] S2, reagent is sequentially put into the flow channel 105a

[0177] In this detection method, the reagent is sequentially put into the flow channel 105a as follows:

[0178] The reagent B is put in to activate the electrode, the sample mixture is put in, the reagent D is put in to clean the flow channel 105a and the magnetic beads, the electrochemiluminescence detection is performed through the reagent B, the reagent D is put in to clean the flow channel 105a and the magnetic beads, the gas bubble is put in to isolate the reagent D and the reagent A, and the chemiluminescence detection is performed through the reagent A.

[0179] In addition to the above two methods, the device provided by the application can also perform the following two detections, which are two-step sample addition. Compared with the one-step combined detection and one-step sample addition provided by the application, the convenience is slightly worse.

[0180] I. The first method (two-step combined detection: the sample and the detection reagent are put in at different times):

[0181] First, chemiluminescence detection is performed, and then electrochemiluminescence detection is performed, and the detection mode is specifically as follows:

[0182] The reagents and part of the cleaning operations involved in this detection method refer to the specific contents described in the first method, and will not be repeated here.

[0183] S1, sample mixing reaction

[0184] The sample to be tested and the first reagent are mixed uniformly in the reaction cup 106, and then are put into the reaction module for sufficient reaction. Then, the streptavidin-coated magnetic beads are added, mixed, and then are put into the reaction module for sufficient reaction. After the reaction, the chemiluminescence mixed sample is obtained, in which the first binding molecule and the first measured substance are combined to the magnetic beads.

[0185] The sample to be tested and the second reagent are mixed uniformly in the reaction cup 106, and then are put into the reaction module for sufficient reaction. Then, the streptavidin-coated magnetic beads are added, mixed, and then are put into the reaction module for sufficient reaction. After the reaction, the electrochemiluminescence mixed sample is obtained, in which the second binding molecule and the second measured substance are combined to the magnetic beads.

[0186] S2, the reagents are sequentially introduced into the flow channel 105a

[0187] The specific process is shown in the following table: Figure 2 In the detection method, the reagent introduction sequence is as follows:

[0188] Reagent B is introduced to activate the electrode, chemiluminescence mixed sample is introduced, reagent D is introduced to clean the flow channel 105a and the magnetic beads, a bubble isolation reagent D and reagent A are introduced, reagent A is introduced for chemiluminescence detection, reagent D is introduced to clean the flow channel 105a, electrochemiluminescence mixed sample is introduced, reagent B is introduced to clean the magnetic beads and perform electrochemiluminescence detection.

[0189] Second (two-step combined detection: sample and detection reagent are introduced in two steps):

[0190] First, electrochemiluminescence detection is performed, and then chemiluminescence detection is performed. The detection method is as follows:

[0191] The reagents and part of the cleaning operation involved in the detection method are described in detail in the first method of one-step combined detection, which will not be described in detail here.

[0192] S1, sample mixing reaction

[0193] The sample to be tested and the first reagent are mixed uniformly in the reaction cup 106, and then are put into the reaction module for sufficient reaction. Then, the streptavidin-coated magnetic beads are added, mixed, and then are put into the reaction module for sufficient reaction. After the reaction, the chemiluminescence mixed sample is obtained, in which the first binding molecule and the first measured substance are combined to the magnetic beads.

[0194] The sample to be tested and the second reagent are mixed uniformly in the reaction cup 106, and then are put into the reaction module for sufficient reaction. Then, the streptavidin-coated magnetic beads are added, mixed, and then are put into the reaction module for sufficient reaction. After the reaction, the electrochemiluminescence mixed sample is obtained, in which the second binding molecule and the second measured substance are combined to the magnetic beads.

[0195] S2, the reagents are sequentially introduced into the flow channel 105a

[0196] In the detection method, the reagent input sequence is as follows:

[0197] Input reagent B, activate the electrode by electrifying the electrode, input electrochemiluminescence mixed sample, input reagent D to clean the flow channel 105a and magnetic beads, input reagent B for electrochemiluminescence detection, input chemiluminescence mixed sample, input reagent D to clean the flow channel 105a and magnetic beads, input bubble isolation reagent D and reagent A, input reagent A for chemiluminescence detection.

[0198] The following will be described in conjunction with specific examples.

[0199] The embodiments of the present application will be described in detail in conjunction with examples. It should be understood that the examples are only used to illustrate the present application and not intended to limit the scope of the present application. The experimental methods in the following examples without specific conditions are preferred to refer to the guidance given in the present application, and can also be according to the experimental manual or conventional conditions in the art, or according to the conditions suggested by the manufacturer, or according to the known experimental methods in the art.

[0200] In the following specific examples, the amount of raw material components may have slight deviations within the weighing accuracy range, if not otherwise specified. The temperature and time parameters allow for acceptable deviations caused by instrument testing accuracy or operation accuracy.

[0201] Example 1

[0202] The embodiment uses the thyroid stimulating hormone TSH project and the thyroid globulin Tg project combined reagent kit of the thyroid function package as an example. First, the TSH project and the Tg project combined reagent kit components include quantum dot labeled TSH antibody (concentration of 2.0 μg / mL) and trispyridine ruthenium labeled Tg antibody. The specific components of the first reagent are as follows: 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, and pH value is 7.4. The specific components of the second reagent are as follows: trispyridine 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, and 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 method reaction, the mixture is added to the reaction container (reaction cup 106) together with biotin labeled TSH antibody and biotin labeled Tg antibody by the reagent loading device. After the sample loading device adds the sample to be tested to the reaction cup 106 and mixes, the reaction cup 106 is placed in the reaction module for reaction for 9 minutes. After the reaction is completed, the reagent loading device adds streptavidinized magnetic beads, mixes, and then places in the reaction module for reaction for another 9 minutes. The average particle size of the magnetic beads used is 1 μm-3.0 μm. The quantum dot labeled TSH antibody is from Puhuitongke Technology, and the quantum dot is a CdTe quantum dot. The trispyridine ruthenium labeled Tg antibody is from Puhuitongke Technology.

[0203] After the reaction is completed, the sample to be tested and the reagent form a combination, 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 is attached to the pit below the light-emitting area at the bottom of the measuring pool 105. As shown in FIG. 6, the combination slowly flows through the reaction flow channel 105a of the measuring pool 105 under the timing control, and the magnetic beads in the reaction combination are adsorbed to the bottom of the flow channel 105a above the magnetic adsorption member 104, and the flow rate is 20 μL / S. Figure 4

[0204] The detection needle 102 sucks the reagent D, repeatedly rinses the magnetic beads, and the substances not combined with the magnetic beads are taken away by the reagent D under the action of the fluid.

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

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

[0207] After the quantum dots complete light emission, reagent B with a magnetic bead washing effect is introduced into the flow channel 105a, and the reagent B does not react with the quantum dots and the tris (2-phenylpyridine) ruthenium under the condition of no power supply. During the process of introducing the reagent B into the flow channel 105a, the magnetic adsorption member 104 is attached to the bottom of the measuring cell 105. The reagent B is filled at a speed of 200 μL / s, and after the magnetic bead washing is completed, the measuring cell 105 is powered, and in this case, 1.4 V is used to excite electrochemiluminescence (the tris (2-phenylpyridine) ruthenium and tripropylamine are usually excited at a voltage of 1.2 V to 1.5 V). A narrow band filter 107 is installed at the front end of the photon detection and collection device 103. The light emission is detected by the corresponding optical detection module 103, and the Tg signal intensity can be obtained.

[0208] After the light emission is completed, the electrochemical luminescence measuring cell 105, the detection needle 102 and the pipeline are washed by the reagent C, and a step change voltage is applied during the washing process to ensure the washing effect.

[0209] Example 2

[0210] The embodiment is basically the same as example 1, and the difference is that the TSH project and Tg project combined reagent kit components 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); 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 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-20 0.5%wt, Triton X-100 0.5%wt, BSA 3%wt, ProClin-300 0.2%wt, pH value is 7.4. The CdTe quantum dot labeled TSH antibody and the CuSe quantum dot labeled Tg antibody are both from Puhuitongke Technology Co., Ltd.

[0211] Example 3

[0212] The reagent composition of this embodiment is the same as that of Example 1, except that a two-step combined detection method is used, i.e. the sample to be detected is mixed with the first reagent and the second reagent respectively in the reaction cup 106, and then is placed in the reaction module for sufficient reaction, after which streptavidin-coated magnetic beads are added, and after mixing, the mixture is placed in the reaction module for sufficient reaction. After the reaction, chemiluminescence mixed sample and electrochemiluminescence mixed sample are obtained respectively. The two mixed samples are sequentially introduced into the flow channel 105a, and the specific flow process is shown in Figure 3

[0213] The luminescence wavelength of the CdTe quantum dots in this example is 785 nm, and the luminescence wavelength of the CdSe quantum dots is 550 nm. A wavelength narrowband filter 107 is used for filtering before the optical detection module 103; according to the luminescence intensity, the TSH and Tg signal intensities can be obtained respectively.

[0214] Example 4

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

[0216] Example 5

[0217] This embodiment is basically the same as Example 1, except that electrochemiluminescence is performed first, followed by chemiluminescence, and the reagent introduction sequence is: reagent B electrode is introduced to activate the electrode, sample mixture is introduced, reagent D is introduced to clean the flow channel 105a and the magnetic beads, electrochemiluminescence detection is performed through 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 chemiluminescence detection is performed through reagent A.

[0218] Example 6

[0219] This embodiment is basically the same as Example 5, except that the specific components of the second reagent are: Tg antibody labeled with CdSe quantum dots 0.5 μg / mL, PBS 0.05 M, 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 example is basically the same as Example 5, except that a two-step combined detection method is used, i.e. the sample to be tested is mixed with the first reagent and the second reagent respectively in the reaction cup 106, and then put into the reaction module for sufficient reaction, after which the streptavidin-coated magnetic beads are added, mixed, and then put into the reaction module for sufficient reaction. After the reaction is completed, the chemiluminescence mixed sample and the electrochemiluminescence mixed sample are obtained respectively, and the two mixed samples are sequentially introduced into the flow channel 105a, and the electrochemiluminescence is performed first and then the chemiluminescence.

[0222] Example 8

[0223] This example is basically the same as Example 7, except that the specific components of the second reagent are: Tg antibody 0.5 μg / mL labeled with CdSe quantum dots, 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. The Tg is detected by electrochemiluminescence using the same equipment as in Example 1, and the specific content is basically similar to that of Example 1, except that the quantum dot-labeled TSH antibody is not added to the first reagent, but an acridinium ester-labeled TSH antibody is added, and the reagent A is not introduced into the flow channel 105a during testing, but a chemiluminescence co-reaction reagent H2O2 / NaOH solution is introduced.

[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, except that the second reagent is a quantum dot-labeled Tg antibody.

[0228] Comparative Example 3

[0229] This comparative example is a comparative example of Examples 3 and 7. This comparative example is basically the same as Comparative Example 1, except that a two-step combined 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 basically the same as Comparative Example 2, except that a two-step combined detection method is used.

[0232] Test:

[0233] Five different concentrations of TSH / Tg serum samples were prepared. The different concentrations of standard samples were detected by the detection method of examples 1-8. Each sample was determined three times, and the test values 1-3 were obtained, and the average was obtained. The detection results are shown in Table 1. Figure 1 and Table 1.

[0234] The master curve was established by using the calibration samples with traceable values in examples 1-8 and comparative examples 1-4, and two standard samples were tested, and the results are shown in Table 2.

[0235] Table 1 Test sample results of different examples

[0236]

[0237]

[0238] Table 2 Test standard sample results of different examples and comparative examples

[0239]

[0240]

[0241]

[0242] From Table 1, it can be seen that different examples all have very high signal-to-noise ratio, which shows that whether quantum dot chemiluminescence, quantum dot electrochemiluminescence, or trispyridine ruthenium electrochemiluminescence has very high detection capability.

[0243] From Table 2, it can be seen that when using one-step joint detection method, quantum dot chemiluminescence-quantum dot electrochemiluminescence or trispyridine ruthenium electrochemiluminescence has higher accuracy than ordinary chemiluminescence. The reason may be that the ordinary chemiluminescence has a light-emitting wavelength of 420-470 nm, which is the same as the absorption wavelength of quantum dots or trispyridine ruthenium, resulting in energy transfer during light emission and affecting the accuracy of detection.

[0244] The TSH chemiluminescence signal-concentration relationship curve and the Tg electrochemiluminescence signal-concentration relationship curve of each example are shown in Figure 5 From the figure, it can be seen that the higher the concentration of TSH and Tg, the higher the light intensity.

[0245] In summary, the chemiluminescence-electrochemiluminescence co-detection device provided by the present application can realize chemiluminescence and electrochemiluminescence in the same device. When multiple substances in the sample to be detected need to be analyzed and detected, both chemiluminescence detection and electrochemiluminescence detection are required, which greatly improves the convenience of detection. The chemiluminescence detection in the flow channel can reduce the individual differences caused by the sample container compared with the conventional chemiluminescence detection, and the accuracy is higher.

[0246] The co-detection device provided in the preferred embodiment of the present application comprises a control system, and the control system is built-in with a time sequence control program, which realizes automatic sampling by preset time nodes, so as to realize automatic detection of chemiluminescence and electrochemiluminescence.

[0247] The chemiluminescence-electrochemiluminescence detection method provided by the present application can realize electrochemiluminescence detection and chemiluminescence detection in the same device, which greatly improves the convenience of detection when the sample needs both luminescence detection and electrochemiluminescence detection; and compared with the conventional chemiluminescence detection, the chemiluminescence detection in the flow channel can reduce the individual difference caused by the sample container, and is more accurate.

[0248] The detection method provided in the preferred embodiment of the present application combines chemiluminescence markers (quantum dots) and electrochemiluminescence markers (trispyridine ruthenium) and the like, and realizes time and wavelength resolution multiplex detection through chemiluminescence and electrochemiluminescence.

[0249] The detection method provided in the preferred embodiment of the present application uses quantum dots as chemiluminescence and electrochemiluminescence markers, which are combined with chemiluminescence co-reagents and electrochemiluminescence co-reagents respectively, to realize chemiluminescence and electrochemiluminescence at different time nodes.

[0250] When the chemiluminescence markers and the electrochemiluminescence markers are both quantum dots, the same kind of quantum dots can be selected, or different kinds of quantum dots can be selected. When the same kind of quantum dots are selected, the same quantum dots can realize detection of two analytes in one measuring pool through chemiluminescence and electrochemiluminescence; when different kinds of quantum dots are selected, multiple quantum dots can realize time and wavelength resolution multiplex detection through chemiluminescence and electrochemiluminescence.

[0251] The above is only a specific embodiment of the present application, but the protection scope of the present application is not limited thereto, and any person skilled in the art can easily think of changes or replacements within the technical range disclosed by the present application, which should be covered in the protection scope of the present application. Therefore, the protection scope of the present application should be subject to the protection scope of the claims.

Claims

1. A chemical-electrochemiluminescence co-detection apparatus, characterized in that, The device comprises a reaction cup, a plurality of reagent containers, a detection needle, a sample injection control module, a measuring pool, an electrode assembly, a magnetic attraction accessory, and at least one set of optical detection module. The reaction cup is used to hold a sample mixture, which comprises a combination of at least two target substances in the sample and at least two binding molecules, which are specifically bound to the at least two target substances and adsorbed onto magnetic beads, at least one of the at least two binding molecules is a chemiluminescent label, and at least one of the at least two binding molecules is an electrochemiluminescent label. At least part of the plurality of reagent containers are used to hold a plurality of luminescent reagents, including at least one chemiluminescent co-reagent and at least one electrochemiluminescent co-reagent. The detection needle is used to suck the sample mixture from the reaction cup into the flow channel of the measuring pool and to suck the corresponding luminescent reagent from the plurality of reagent containers into the flow channel of the measuring pool. The sample injection control module controls the detection needle to suck the sample mixture into the reaction cup and the plurality of reagent containers. The measuring pool provides a reaction space for chemiluminescence and electrochemiluminescence. The electrode assembly is arranged with the measuring pool and is used to excite the electrochemiluminescent label to emit light after being powered on. The magnetic attraction accessory is arranged near the measuring pool and is used to magnetically attract the magnetic beads in the flow channel of the measuring pool. The at least one set of optical detection module is used to detect and collect the luminescent intensity of the luminescent label. The chemiluminescence-electrochemiluminescence co-detection device is configured as follows: The sample injection control module controls the detection needle to suck the sample mixture into the flow channel of the measuring pool. After the combination is adsorbed by the magnetic attraction accessory, the sample injection control module controls the detection needle to suck the luminescent reagent into the flow channel of the measuring pool. When the sucked luminescent reagent is the chemiluminescent co-reagent, the chemiluminescent co-reagent combines with the corresponding luminescent label to realize chemiluminescence, and the luminescent intensity is detected and collected by the at least one set of optical detection module; when the sucked luminescent reagent is the electrochemiluminescent co-reagent, the electrode assembly is powered on, the electrochemiluminescent co-reagent combines with the corresponding luminescent label to realize electrochemiluminescence, and the luminescent intensity is detected and collected by the at least one set of optical detection module. The chemiluminescence-electrochemiluminescence co-detection device further comprises a control system. The control system is communicatively connected with the sample injection control module and the electrode assembly, and a time sequence control program is built in the control system, which is configured to instruct the sample injection control module to control the detection needle to suck the corresponding reagent from different reagent containers at different time nodes. When the current time node is the time node for sucking the electrochemiluminescent co-reagent, the control system issues an instruction to the electrode assembly to power on, and after the electrochemiluminescent reaction is completed, the control system controls the electrode assembly to power off. ​ 2. The chemiluminescent-electrochemiluminescent co-detection apparatus according to claim 1, wherein The timing control program is configured to control the sample injection control module to make the detection needle first sample the chemiluminescence co-reagent to realize chemiluminescence and then sample the electrochemiluminescence co-reagent to realize electrochemiluminescence. Alternatively, the timing control program is configured to control the sample injection control module to make the detection needle first sample the electrochemiluminescence co-reagent to realize electrochemiluminescence and then sample the chemiluminescence co-reagent to realize chemiluminescence.

3. The chemiluminescent-electrochemiluminescent co-detection apparatus according to claim 1, wherein Part of the plurality of reagent containers are used to contain neutral cleaning liquid; the timing control program is configured to: control the sample injection control module to make the detection needle sample the sample mixture, suck the neutral cleaning liquid, sample the chemiluminescence co-reagent and sample the electrochemiluminescence co-reagent in sequence; or control the sample injection control module to make the detection needle sample the sample mixture, suck the neutral cleaning liquid, sample the electrochemiluminescence co-reagent and sample the chemiluminescence co-reagent in sequence.

4. The chemiluminescent-electrochemiluminescent co-detection apparatus according to claim 3, wherein The detection needle is also used to suck air into the flow channel of the measuring pool to form bubbles in the flow channel; the timing control program is configured to: control the sample injection control module to make the detection needle sample the sample mixture, suck the neutral cleaning liquid, suck air, sample the chemiluminescence co-reagent and sample the electrochemiluminescence co-reagent in sequence; or control the sample injection control module to make the detection needle sample the sample mixture, suck the neutral cleaning liquid, suck air, sample the electrochemiluminescence co-reagent and sample the chemiluminescence co-reagent in sequence.

5. The chemiluminescent-electrochemiluminescent co-detection apparatus according to claim 1, wherein The bottom of the outer wall of the measuring pool is provided with a receiving groove for receiving at least part of the magnetic adsorption member; the magnetic adsorption member comprises the following features (1) or (2): (1) the magnetic adsorption member is a permanent magnet, and the chemiluminescence-electrochemiluminescence co-detection device further comprises a movement control module, which controls the magnetic adsorption member to move into the receiving groove to adsorb the magnetic beads at the beginning of luminescence detection and controls the magnetic adsorption member to move out of the receiving groove to release the adsorption of the magnetic beads by the magnetic adsorption member after the end of luminescence detection; (2) the magnetic adsorption member is an electromagnet, which is configured to be powered on at the beginning of luminescence detection and powered off after the end of luminescence detection.

6. The chemiluminescent-electrochemiluminescent co-detection apparatus according to claim 1, wherein The number of optical detection modules is one set, and the luminescence wavelengths of all luminescent markers used for detection can be detected and collected by the optical detection module; Alternatively, the number of optical detection modules is multiple sets, and the luminescent markers used for detection are multiple, the luminescent markers of different luminescent wavelengths are different, and the luminescent intensities of the lights emitted by the luminescent markers of different luminescent wavelengths are respectively detected and collected by the corresponding optical detection modules.

7. The chemiluminescence-electrochemiluminescence co-detection apparatus according to claim 1, wherein, The upper wall of the measuring pool is provided with a transparent window, and each set of optical detection modules is arranged above the transparent window, and a corresponding wavelength filter is arranged between each set of optical detection modules and the transparent window.

8. The chemiluminescent-electrochemiluminescent co-detection apparatus according to claim 7, wherein The filter is a narrow-band filter lens.

9. A chemiluminescence-electrochemiluminescence detection method, characterized in that, The chemical-electrochemical luminescence co-detection device is implemented by adopting the method according to any one of claims 1-8, and comprises the following steps. A sample mixture is introduced into a flow channel of a measuring pool, the sample mixture comprising a complex obtained by sufficient reaction of a sample to be measured, at least one first binding molecule, at least one second binding molecule, and magnetic beads, and the complex being adsorbed on the bottom of the flow channel when passing near a magnetic adsorption member arranged outside the flow channel; A plurality of luminescent reagents are introduced into the flow channel of the measuring pool, each luminescent reagent being introduced at a different time node, and the plurality of luminescent reagents comprising at least one chemical luminescence co-reagent and at least one electrochemical luminescence co-reagent; Each of the first binding molecules is a molecule labeled by a chemical luminescence label, when the number of the first binding molecules in the sample mixture is a plurality, the plurality of the first binding molecules are molecules labeled by different chemical luminescence labels respectively, and each of the first binding molecules is a substance capable of specifically binding to a corresponding type of measured substance in the sample to be measured and being adsorbed on the magnetic beads; Each of the second binding molecules is a molecule labeled by an electrochemical luminescence label, when the number of the second binding molecules in the sample mixture is a plurality, the plurality of the second binding molecules are molecules labeled by different electrochemical luminescence labels respectively, and each of the second binding molecules is a substance capable of specifically binding to a corresponding type of measured substance in the sample to be measured and being adsorbed on the magnetic beads; The at least one chemical luminescence co-reagent is combined with the luminescent labels on the at least one first binding molecule one by one to realize chemical luminescence, and the corresponding luminescent intensity is detected and collected by a corresponding optical detection module; When the electrochemical luminescence co-reagent is introduced into the flow channel, an electrode assembly in the measuring pool is powered on, the at least one electrochemical luminescence co-reagent is combined with the luminescent labels on the at least one second binding molecule one by one to realize electrochemical luminescence, and the corresponding luminescent intensity is detected and collected by a corresponding optical detection module.

10. The luminescence detection method according to claim 9, characterized in that Before the different luminescent reagents are introduced into the flow channel of the measuring pool, a neutral cleaning liquid is also introduced into the flow channel of the measuring pool.

11. The luminescence detection method of claim 9, wherein, When the complex is adsorbed on the bottom of the flow channel by the magnetic adsorption member, a neutral cleaning liquid is introduced into the flow channel to wash away free substances in the sample mixture that are not combined with the magnetic beads, and then the luminescent reagents are introduced.

12. The luminescence detection method of claim 11, wherein, After the free substances not combined with the magnetic beads are washed away, before the luminescent reagents are introduced, a gas bubble is also introduced into the flow channel to isolate the neutral cleaning liquid and the luminescent reagents.

13. The luminescence detection method of claim 11, wherein, The flow rate of the sample mixture in the flow channel is 10 μL / S-30 μL / S, the flow rate of the chemical luminescence co-reagent in the flow channel is 100 μL / S-500 μL / S, and the flow rate of the electrochemical luminescence co-reagent in the flow channel is 100 μL / S-200 μL / S.

14. The luminescence detection method according to any one of claims 10-13, wherein the neutral cleaning solution is a solution containing a phosphate salt and a detergent, the concentration of the phosphate salt is 10 mM-500 mM, and the mass percentage of the detergent is 0.05%-1%.

15. The luminescence detection method according to any one of claims 9 to 13, wherein, Further comprising at least one of the following features (1)-(5); (1) after all luminescence detection is completed, further comprising sequentially introducing a basic cleaning solution and the electrochemiluminescence co-reagent into the flow channel, the basic cleaning solution is a solution containing a detergent and a strong base, the mass percentage of the detergent is 0.05%-1%, the detergent is at least one selected from Tween, Triton and polycarboxylic alcohol, the strong base is at least one selected from KOH and NaOH, and the pH of the basic cleaning solution is 12-14; (2) the electrochemiluminescence label is terpyridine ruthenium or quantum dots; (3) the chemiluminescence label is quantum dots; (4) before electrochemiluminescence detection is performed by using the device, an electrode activation operation is performed, and when the electrode activation operation is performed, the electrochemiluminescence co-reagent is introduced into the flow channel; (5) the electrochemiluminescence co-reagent contains a surfactant for cleaning the surface of magnetic beads, the surfactant is at least one selected from Tween, Triton and polycarboxylic alcohol, and the mass percentage of the surfactant is 0.05%-1%.

16. The luminescence detection method according to any one of claims 9 to 13, wherein, The electrochemiluminescence label is terpyridine ruthenium, and the electrochemiluminescence co-reagent corresponding to the terpyridine ruthenium is a solution containing tripropylamine.

17. The luminescence detection method according to claim 16, wherein the solution containing tripropylamine is a tripropylamine phosphate solution.

18. The luminescence detection method according to claim 17, wherein the concentration of tripropylamine in the tripropylamine phosphate solution is 0.1 M-0.2 M, and / or the concentration of the phosphate salt in the tripropylamine phosphate solution is 10 mM-500 mM.

19. The luminescence detection method according to claim 17, wherein the concentration of terpyridine ruthenium in the tripropylamine phosphate solution is 0.01 nM-5 μM.

20. The luminescence detection method according to any one of claims 9 to 13, wherein, The chemiluminescence label is quantum dots, and the chemiluminescence co-reagent corresponding to the quantum dots is a solution containing potassium permanganate.

21. The luminescence detection method of claim 20, wherein, The solution containing potassium permanganate is a potassium permanganate phosphate solution.

22. The luminescence detection method of claim 21, wherein, The concentration of potassium permanganate in the potassium permanganate phosphate solution is 1 mM-3 mM, and / or the concentration of the phosphate salt is 1 mM-100 mM.

23. The luminescence detection method of claim 21, wherein, The concentration of the quantum dots in the potassium permanganate phosphate solution is 0.5 μM-1.5 μM.

24. The luminescence detection method according to any one of claims 9 to 13, wherein, The first binding molecule is a quantum dot-labeled first antibody to be detected, and the chemiluminescence co-reagent is a solution containing potassium permanganate; The second binding molecule is a terpyridine ruthenium-labeled second antibody to be detected, and the electrochemiluminescence co-reagent is a solution containing tripropylamine.

25. The luminescence detection method according to any one of claims 9 to 13, wherein, The chemiluminescence label and the electrochemiluminescence label are both quantum dots; The chemiluminescence co-reagent is a solution containing potassium permanganate, and the electrochemiluminescence co-reagent is a solution containing tripropylamine.

26. The luminescence detection method of claim 25, wherein, The quantum dots are CdTe, CdSe or CuInS2@ZnS.

27. The luminescence detection method according to any one of claims 9 to 13, wherein, The sample mixture is obtained by mixing a sample to be tested, a first reagent, a second reagent and magnetic beads; The first reagent comprises quantum dot-labeled first detection antibody 0.05 mg / L-50.0 mg / L, buffer salt solution 1 mmol / L-100 mmol / L, surfactant 0.5 g / L-50 g / L, preservative 0.05 g / L-5 g / L, and the pH value of the first reagent is 5.0-9.

0. The second reagent comprises terpyridine ruthenium-labeled second detection antibody 0.05 mg / L-50.0 mg / L, buffer salt solution 1 mmol / L-100 mmol / L, surfactant 0.5 g / L-50 g / L, preservative 0.05 g / L-5 g / L, and the pH value of the first reagent is 5.0-9.

0. Alternatively, the second reagent comprises quantum dot-labeled second detection antibody 0.05 mg / L-50.0 mg / L, buffer salt solution 1 mmol / L-100 mmol / L, surfactant 0.5 g / L-50 g / L, preservative 0.05 g / L-5 g / L, and the pH value of the first reagent is 5.0-9.

0.

28. The luminescence detection method of claim 27, wherein, The buffer salt solution comprises a combination of one or more of PBS, PB, PIPES, MES, HEPES, TRIS, HEPPS and Bis-Tris; The surfactant comprises a combination of one or more of polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monooleate, polyethylene glycol tert-octyl phenyl ether, polyethylene glycol nonyl phenyl ether and fatty alcohol polyoxyethylene ether; The preservative comprises a combination of one or more of ProClin-300, ProClin-950, gentamicin and 5-bromo-5-nitro-1,3-dioxane.

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