Fully automated single-molecule immunoassay analyzer

Abstract

A fully automated single‑molecule immunoassay analyzer, comprising a sample adding module (1), a reagent module (4), an incubation temperature control module (7), and an optical detection module. The incubation temperature control module (7) comprises: a turntable (85) provided with multiple circumferentially arranged bearing positions (86) for reaction vessels (57); a motor (81) and a first rotating shaft (82), the turntable (85) being fixed on the first rotating shaft (82), and the motor (81) driving the first rotating shaft (82) to rotate; a heating component and a temperature sensor disposed on the turntable (85); an enclosure (84) and an upper cover (87), the enclosure (84) surrounding the turntable (85), and the upper cover (87) being disposed at an upper opening of the enclosure (84); and a mixing mechanism (5) disposed below the turntable (85), wherein when the turntable (85) rotates and a reaction vessel (57) at any of the bearing positions (86) is above the mixing mechanism (5), the mixing mechanism (5) lifts the reaction vessel (57) and rotates.

Description

Fully automatic single molecule immunoassay analyzer Technical Field

[0001] The present invention relates to immunoassay, in particular to a full-automatic single molecule immunoassay analyzer. Background Art

[0002] The fully automatic single-molecule immunoassay analyzer mainly detects neural markers, myocardial markers, inflammatory markers, tumor markers, infection markers, etc. It detects trace concentrations of samples by detecting single molecules. The single-molecule detection system uses the classic double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) to achieve extremely low-level protein quantitative detection. The traditional ELISA reaction system is relatively large (100μL), and requires millions of molecules to generate tens of millions of fluorophore signals before it can be detected by the enzyme reader. Therefore, the traditional ELISA method can only detect signals above the picogram range and uses an analog algorithm. The biggest feature of single-molecule detection is that it can detect single molecules with extremely high detection sensitivity, usually at the femtogram level (fg / mL), which is 1000 times more sensitive than the traditional ELISA method.

[0003] To meet the demand for ultrasensitive detection, domestic and international manufacturers have developed various single-molecule immunoassay methods. The main single-molecule immunoassay instruments currently on the market include the Simoa-HD-1 and HD-X from Quanterix (USA). However, due to their design limitations, the Simoa-HD-1 and HD-X are relatively large. The HD-X measures 135 × 60 × 160 cm, making it quite bulky.

[0004] When the fully automatic single-molecule fluorescence immunoassay analyzer is working, the instrument performs operations such as sample addition, mixing, reagent addition, and magnetic separation around the reaction cup. The grabbing of the reaction cup is the link connecting various processes, and the quality of its function directly affects whether the entire machine can operate normally.

[0005] The gripping device of the reaction cup needs to meet the following requirements:

[0006] 1. Due to the limitation of the overall layout space, the machine needs to have a compact structure and small size.

[0007] 2. The wall of the reaction cup is thin and easy to deform, so the clamping force of the clamping device needs to be flexible and controllable to prevent the reaction cup from being deformed by force.

[0008] 3. Reliable placement and removal, and has a certain degree of compatibility with defects in the reaction cup, such as slight deformation of the cup edge.

[0009] For similar cuvette grabbing problems, the solutions generally adopted by domestic and foreign manufacturers are:

[0010] An optional electromagnet drives a cam as the driving mechanism. Springs maintain the clamping tendency on the left and right swing arms, each equipped with a left and right clamping jaw. The jaws engage in a three-jaw chuck-like structure. A motor drives the cam to open the slider, and springs maintain the clamping tendency. The slider requires two guide rails / guide shafts for support. This solution presents the following problems:

[0011] 1. The clamping structure similar to the three-jaw chuck is not stable enough, and the cup is easy to tilt during high-speed movement.

[0012] 2. The support layout of the guide rail / guide shaft is not compact enough and takes up a lot of space.

[0013] 3. It is not compatible with defects of the reaction cup, such as slight deformation of the cup edge, which is prone to cup sticking failure. Summary of the Invention

[0014] In order to solve the deficiencies in the above-mentioned prior art solutions, the present invention provides a fully automatic single molecule immunoassay analyzer.

[0015] The purpose of the present invention is achieved through the following technical solutions:

[0016] The fully automatic single molecule immunoassay analyzer includes a sample loading module, a reagent module, and an incubation temperature control module; the incubation temperature control module includes:

[0017] A tray having a plurality of cuvette carrying positions distributed along a circumference;

[0018] A motor and a rotating shaft, the tray is fixed on the rotating shaft, and the motor drives the rotating shaft to rotate;

[0019] a heating component and a temperature sensor, wherein the heating component and the temperature sensor are arranged on the tray;

[0020] An enclosure and an upper cover, wherein the enclosure is arranged around the tray, and the upper cover is arranged at the upper end opening of the enclosure;

[0021] The mixing mechanism is arranged on the lower side of the tray. When the tray rotates and a reaction cup at any carrying position is on the upper side of the mixing mechanism, the mixing mechanism lifts the reaction cup and rotates.

[0022] Compared with the prior art, the present invention has the following beneficial effects:

[0023] 1. High integration and small size;

[0024] The mixing mechanism and incubation temperature control module are combined together to significantly reduce the instrument volume;

[0025] 2. Mix well;

[0026] During the forward and reverse rotation of the rotating arm, the moving part is driven to move up and down and rotate at the same time, which better mixes the liquid in the container and has a good mixing effect.

[0027] In order to make the moving parts move up and down stably, a counterweight is set on the moving parts to increase the inertia;

[0028] 3. Simple and stable structure;

[0029] Only a combination of a driving unit, a rotating shaft, a rotating arm, a bearing member and a moving member is required, so that when the rotating arm rotates, the bearing member follows the moving member and moves spirally along the central axis of the rotating shaft, thereby achieving the purpose of mixing the liquid in the container;

[0030] The first rotating arm and the second rotating arm are connected by a first elastic member. The rotation of the cam realizes the separation and approach of the first rotating arm and the second rotating arm, thereby clamping and releasing the container. The clamping force is controlled by the first elastic member such as a spring. The structure is compact and deformation of the container is avoided.

[0031] The clamping jaws adopt arc-shaped groove design, and there are four lines of contact between the container and the clamping jaws, which ensures stable clamping;

[0032] 4. Good working performance;

[0033] When the first and second rotating arms rotate, the design of the centrosymmetric cam ensures that the two rotating arms rotate at the same angle, and the trajectory of the two jaw centers and the cam center are on the same arc, which significantly reduces the difficulty of calibration.

[0034] A notch is set on the arc groove of the clamping jaw to prevent the cup from sticking;

[0035] The second elastic member always ensures that the first rotating arm presses the cam, thereby ensuring the certainty of the spatial position of the clamping jaw. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] The disclosure of the present invention will become more easily understood with reference to the accompanying drawings. Those skilled in the art will readily appreciate that these drawings are merely used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention.

[0037] FIG1 is a front view of a fully automatic single molecule immunoassay analyzer according to an embodiment of the present invention;

[0038] FIG2 is a schematic structural diagram of an incubation temperature control module according to an embodiment of the present invention;

[0039] FIG3 is a schematic structural diagram of an incubation temperature control module according to an embodiment of the present invention.

[0040] FIG4 is a schematic structural diagram of a mixing mechanism according to an embodiment of the present invention;

[0041] FIG5 is a schematic structural diagram of a sample loading module according to an embodiment of the present invention.

[0042] FIG6 is a schematic structural diagram of a clamping mechanism according to an embodiment of the present invention;

[0043] FIG7 is a partial schematic diagram of a clamping mechanism according to an embodiment of the present invention;

[0044] FIG8 is a schematic structural diagram of a clamping jaw according to an embodiment of the present invention. DETAILED DESCRIPTION

[0045] Figures 1-8 and the following description describe optional embodiments of the present invention to teach those skilled in the art how to implement and reproduce the present invention. In order to explain the technical solution of the present invention, some conventional aspects have been simplified or omitted. Those skilled in the art will understand that variations or replacements derived from these embodiments will be within the scope of the present invention. Those skilled in the art will understand that the following features can be combined in various ways to form multiple variations of the present invention. Thus, the present invention is not limited to the following optional embodiments, but is only limited by the claims and their equivalents.

[0046] Example 1:

[0047] The fully automatic single molecule immunoassay analyzer according to an embodiment of the present invention, as shown in FIG1 , comprises:

[0048] Sample adding module 1, consumables area 2, sample area 3, incubation temperature control module 7 and reagent module 4.

[0049] As shown in FIG2-FIG3, the incubation temperature control module 7 includes:

[0050] A tray 85 having a plurality of cuvettes 57 carrying positions 86 distributed along a circumference thereof;

[0051] A motor 81 and a first rotating shaft 82 , wherein the tray 85 is fixed on the first rotating shaft 82 , and the motor 81 drives the first rotating shaft 82 to rotate via a synchronous belt;

[0052] A heating component and a temperature sensor, wherein the heating component and the temperature sensor are arranged on the tray 85;

[0053] A fence 84 and an upper cover 87 , wherein the fence 84 is arranged around the tray 85 , and the upper cover 87 is arranged at the upper end opening of the fence 85 ;

[0054] The mixing mechanism 5 is provided at the lower side of the tray 85 . When the tray 85 rotates and a cuvette 57 at any supporting position 86 is located at the upper side of the mixing mechanism 5 , the mixing mechanism 5 lifts the cuvette 57 and rotates.

[0055] As shown in FIG4 , the mixing mechanism 5 includes:

[0056] A first supporting member 56 , the first supporting member 56 is used to support a reaction cup 57 ;

[0057] A first driving unit 511 and a rotating shaft 512, wherein the first driving unit 511 is used to drive the rotating shaft 512 to rotate forward and reverse in a time-divided manner;

[0058] a third rotating arm 55 , the third rotating arm 55 being disposed on the rotating shaft 512 ;

[0059] The moving member 53 is fixed to the first supporting member 56, and a spiral groove or hole 54 is provided on the moving member 53. The end of the third rotating arm 55 is located in the groove or hole 54. When the third rotating arm 55 rotates, the moving member 53 moves spirally along the central axis direction of the rotating shaft 512.

[0060] In order to enable the moving member 53 to move up and down stably, the rotating shaft 512 is further in the shape of a column or cylinder, the moving member 53 is in the shape of a cylinder, and the outer diameter of the rotating shaft 512 is equal to the inner diameter of the moving member 53.

[0061] In order to make the moving member 53 move up and down stably, further, the first bearing member 56 is arranged on the upper side of the moving member 53, and the mixing mechanism 5 further includes:

[0062] The counterweight 58 is provided on the moving member 53 as shown below.

[0063] As shown in FIG5 , the sample loading module 1 includes a two-dimensional mobile robotic arm and a clamping mechanism 6 ;

[0064] As shown in FIG6-FIG7, the clamping mechanism 6 includes:

[0065] A first clamping jaw 41 and a second clamping jaw 42;

[0066] A second driving unit 31 and a second carrier 21 , wherein the second driving unit 31 is disposed on the second carrier 21 and drives the cam 32 to rotate;

[0067] A first rotating arm 51 and a second rotating arm 52 are disposed on the second supporting member 21 via a second rotating shaft, and the cam 32 is located between the first rotating arm 51 and the second rotating arm 52; the first clamping jaw 41 is disposed on the first rotating arm 51, and the second clamping jaw 42 is disposed on the second rotating arm 52;

[0068] The first elastic member 61 is used to prevent the first rotating arm 51 and the second rotating arm 52 from separating. The cam 32 is clamped between the first rotating arm 51 and the second rotating arm 52, so that when the cam 32 rotates, the first rotating arm 51 and the second rotating arm 52 separate or approach around the second rotating axis, thereby loosening or clamping the container 10.

[0069] In order to ensure the certainty of the spatial position of the first clamping jaw 41, the clamping mechanism 6 further includes:

[0070] The second elastic member 62 pushes the first rotating arm 51 to press the cam 32 .

[0071] In order to reduce the difficulty of calibration, further, during the rotation of the first rotating arm 51 and the second rotating arm 52 , the moving tracks of the centers of the first clamping jaw 41 and the second clamping jaw 42 and the center of the cam 32 are on the same arc.

[0072] In order to achieve the consistency of the rotation angles of the first rotating arm 51 and the second rotating arm 52 , the cam 32 is further centrally symmetrical.

[0073] In order to firmly clamp the container 10 , further, the first clamping jaw 41 and the second clamping jaw 42 respectively adopt arc-shaped grooves, so that the container 10 is clamped using four lines of contact.

[0074] In order to prevent the cup from sticking, further, as shown in FIG8 , the arc-shaped groove is provided with a notch 43 , and the upper end of the clamped container 10 is flush with the notch 43 .

[0075] In order to improve the compactness of the structure, further, the second driving unit 31 is a motor, which is arranged on the upper side of the second carrier 21, and the cam 32, the first rotating arm 51 and the second rotating arm 52 are respectively arranged on the lower side of the second carrier 21.

[0076] Example 2:

[0077] An application example of the fully automatic single molecule immunoassay analyzer according to Example 1 of the present invention.

[0078] In this application example, as shown in Figures 2-3, in the incubation temperature control module 7, a tray 85 has 36 circumferentially distributed positions 86 for a plurality of reaction cups 57. The center of the tray 85 is fixed to a first rotating shaft 82, which is driven by a motor 81 via a synchronous belt. A heating component and a temperature sensor are mounted on the tray 85.

[0079] The enclosure 84 is arranged around the tray 85, and the upper cover 87 is arranged at the upper end opening of the enclosure 84;

[0080] The mixing mechanism 5 is provided at the lower side of the tray 85 . When the tray 85 rotates and a cuvette 57 at any supporting position 86 is located above the mixing mechanism 5 , the mixing mechanism 5 lifts the cuvette 57 and rotates.

[0081] As shown in Figure 4, during mixing, the cuvette 57 is open at its open end. The first drive unit 511 is a motor, and the rotating shaft 512 is cylindrical. Correspondingly, the moving member 53 is cylindrical, with a spiral (through) hole 54 defined in the wall. A third rotating arm 55 is fixed to the rotating shaft 512, perpendicular to its central axis, with its end positioned within the spiral (through) hole 54.

[0082] A first carrier 56 with an open end is fixed to the upper side of the movable member 53 and has an eccentrically arranged concave structure inside for enveloping the reaction cup 57. A counterweight 58 is fixed to the lower side of the movable member 53.

[0083] As shown in FIG6 , in the gripping mechanism 6 , the second drive unit 31 is a motor, which is disposed on the upper side of the second carrier 21 . The motor's shaft passes through the second carrier 21 , and a centrally symmetrical cam 32 is mounted on the motor shaft on the lower side of the second carrier 21 . The container 10 is a cuvette 57 .

[0084] As shown in Figure 7, the first rotating arm 51 and the second rotating arm 52 are respectively arranged on the lower side of the second supporting member 21 through the second rotating shaft. The first elastic member 61 is a spring that tightens the first rotating arm 51 and the second rotating arm 52 so that the first rotating arm 51 and the second rotating arm 52 clamp the cam 32. The second elastic member 62 is a torsion spring that presses the first rotating arm 51 against the cam 32. The first clamping jaw 41 is installed on the inner side of the first rotating arm 51, and the second clamping jaw 42 is installed on the inner side of the second rotating arm 52. In the vertical direction, the second driving unit 31, the second supporting member 21, the cam 32 and the clamping jaw are arranged in sequence. During the rotation of the cam 32, the clamping jaw will not touch.

[0085] As shown in FIG8 , the first clamping jaw 41 and the second clamping jaw 42 both adopt a vertical arc-shaped groove design. A notch 43 is provided on the arc-shaped groove of the first clamping jaw 41 . When clamping the container 10 , the upper end surface of the container 10 is flush with the notch 34 .

[0086] The working mode of the mixing mechanism 5 of this embodiment is:

[0087] When the tray 85 rotates, the cuvette 57 is located on the upper side of the mixing mechanism 5. The first driving unit 511 drives the rotating shaft 512 to rotate in the forward direction, driving the third rotating arm 55 to rotate in the spiral (through) hole 54. The driving member 53 moves upward while rotating, so that the first supporting member 56 carries the cuvette 57 and spirals upward until the third rotating arm 55 is at the lowest point of the spiral (through) hole 54.

[0088] The rotating shaft 512 stops rotating, and under the weight of the first bearing member 56 , the counterweight 58 and the movable member 53 , the movable member 53 moves downward and rotates by itself until the third rotating arm 55 is at the highest point of the spiral (through) hole 54 .

[0089] The reaction cup 57 moves up and down following the first carrier 56 and rotates at the same time, thereby mixing the liquid in the reaction cup 57. When the mixing is completed, the moving member 53 is at the lowest position and does not prevent the reaction cup 57 from rotating with the tray 85.

[0090] The working mode of the clamping mechanism 6 in this embodiment is:

[0091] The second driving unit 31 drives the cam 32 to rotate, and under the pulling force of the first elastic member 61, the first rotating arm 51 and the second rotating arm 52 move away from each other, so that the first clamping jaw 41 and the second clamping jaw 42 release the container 10, as shown in FIG. 7 .

[0092] The second driving unit 31 drives the cam 32 (the minimum diameter must be smaller than the outer diameter of the container 10) to rotate, and under the pulling force of the first elastic member 61, the first rotating arm 51 and the second rotating arm 52 approach each other, so that the first clamping jaw 41 and the second clamping jaw 42 clamp the container 10. There are four lines of contact between the container 10 and the clamping jaws, as shown in Figure 8. Due to the design of the notch 43, the cup rim 11 of the container 10 does not contact the first clamping jaw 41.

Claims

1. A fully automatic single molecule immunoassay analyzer, comprising a sample loading module, a reagent module and an incubation temperature control module; characterized in that: The incubation temperature control module comprises: A tray, wherein the tray has a plurality of cuvette carrying positions distributed along a circumference; A motor and a rotating shaft, the tray is fixed on the rotating shaft, and the motor drives the rotating shaft to rotate; A heating component and a temperature sensor, wherein the heating component and the temperature sensor are arranged on the tray; A fence and an upper cover, wherein the fence is arranged around the tray, and the upper cover is arranged at the upper end opening of the fence; A mixing mechanism is arranged at the lower side of the tray. When the tray rotates and a reaction cup at any carrying position is located at the upper side of the mixing mechanism, the mixing mechanism lifts up the reaction cup and rotates.

2. The fully automatic single molecule immunoassay analyzer according to claim 1, characterized in that: The mixing mechanism comprises: A driving unit and a rotating shaft, wherein the driving unit is used to drive the rotating shaft to rotate forward and reverse in time; A rotating arm, wherein the rotating arm is arranged on the rotating shaft; A supporting member and a moving member, wherein the supporting member is used to support the reaction cup and is fixed on the upper side of the moving member, a spiral groove or hole is provided on the moving member, the end of the rotating arm is located in the groove or hole, and when the rotating arm rotates, the moving member moves spirally along the central axis direction of the rotating shaft.

3. The fully automatic single molecule immunoassay analyzer according to claim 2, characterized in that: The rotating shaft is cylindrical or cylindrical, the moving part is cylindrical, and the outer diameter of the rotating shaft is equal to the inner diameter of the moving part.

4. The fully automatic single molecule immunoassay analyzer according to claim 2, characterized in that: The mixing mechanism also includes: A counterweight is arranged on the moving part.

5. The fully automatic single molecule immunoassay analyzer according to claim 4, characterized in that: The counterweight is arranged on the lower side of the moving member.

6. The fully automatic single molecule immunoassay analyzer according to claim 1, characterized in that: The sample loading module includes a two-dimensional mobile mechanical arm and a clamping mechanism, and the clamping mechanism includes: a first clamping jaw and a second clamping jaw; A driving unit and a bearing, wherein the driving unit is arranged on the bearing and drives the cam to rotate; A first rotating arm and a second rotating arm, wherein the first rotating arm and the second rotating arm are arranged on the bearing member via a rotating shaft, and the cam is located between the first rotating arm and the second rotating arm; the first clamping jaw is arranged on the first rotating arm, and the second clamping jaw is arranged on the second rotating arm; A first elastic member is used to prevent the first rotating arm and the second rotating arm from being separated, and the cam is sandwiched between the first rotating arm and the second rotating arm.

7. The fully automatic single molecule immunoassay analyzer according to claim 6, characterized in that: The clamping mechanism also includes: A second elastic member, wherein the second elastic member pushes the first rotating arm to press the cam.

8. The fully automatic single molecule immunoassay analyzer according to claim 6, characterized in that: During the rotation of the first rotating arm and the second rotating arm, the moving tracks of the centers of the first clamping jaw and the second clamping jaw and the center of the cam are on the same arc.

9. The fully automatic single molecule immunoassay analyzer according to claim 6, characterized in that: The cam is centrally symmetrical.

10. The fully automatic single molecule immunoassay analyzer according to claim 6, characterized in that: The driving unit is a motor, which is arranged on the upper side of the carrier, and the cam, the first rotating arm and the second rotating arm are respectively arranged on the lower side of the carrier.

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