Fully automated biochip workstation and detection methods using it

Abstract

The present disclosure relates to the field of detection devices for biochips, and in particular to a fully automated biochip workstation and a detection method using it; a detection device for biochips comprises: a cavity for holding a test tube vial, the test tube serving to hold a sample to be detected and a biochip, and the cavity having a temperature control function; a liquid storage device for storing a liquid; a pumping device for pumping the liquid in the liquid storage device into the test tube vial, the pumping device being in fluid communication with a liquid separation needle, the liquid separation needle being configured for injecting a mixed liquid into the test tube vial; an imaging device for monitoring the condition of a chip in the test tube vial;and a data processing device that obtains a hybridization result based on the information about the state of the biochip acquired by the imaging device. The workstation provided in this disclosure integrates the response of a biochip and the output of detection results, increases sample processing efficiency, saves labor, and enables the obtaining of stable and reliable results.

Description

[0001] The present disclosure claims priority to Chinese Patent Application No. 201811106959.8, filed on September 21, 2018, in the China Patent Office, entitled “Fully Automatic Biochip Workstation and Detection Method Using the Same,” the entire disclosure of which is incorporated herein by reference. Technical area

[0002] The present disclosure relates to the field of chip hybridization devices, and more particularly to a fully automated biochip workstation and a detection method using the same. Technical background

[0003] Biochip hybridization refers to the reaction of a protein, DNA, or RNA sample separated from a biological sample with a biochip to obtain the sample's sequence information from a probe array immobilized on the biochip. Since the fluorescence background of the slide itself is very low, the biochip can be detected and analyzed using a fluorescence labeling method, achieving advantages such as speed and accuracy. Furthermore, multiple biological samples can be analyzed at once by using a variety of fluorescein labeling methods.

[0004] Currently, some devices related to biochip hybridization have been disclosed, but there is no description yet regarding instruments with integrated functions for nucleic acid amplification and hybridization.

[0005] It is with this in mind that the present disclosure is specifically proposed. Subject of revelation

[0006] An object of the present disclosure includes providing a detection device for biochips, wherein the integration of the reaction of a biochip and the output of detection results is realized, the treatment efficiency of samples is increased, the labor is saved, and stable and reliable results can be obtained.

[0007] The object of the present disclosure also includes providing a detection method for biochips, wherein the entire detection process is automated, high detection efficiency and stable and reliable detection results can be achieved.

[0008] To solve at least one of the above tasks or possibly additional tasks, the following technical solution is used: A biochip detection device comprises: a cavity for supporting a test tube vial, the test tube vial for accommodating a sample to be detected and a biochip, and the cavity has a temperature control function; a liquid storage device for storing a liquid; a pumping device for pumping the liquid in the liquid storage device into the test tube vial, the pumping device being in fluid communication with a liquid separation needle, the liquid separation needle being configured to inject a mixed liquid into the test tube vial; an imaging device for monitoring the condition of a chip in the test tube vial; and a data processing device that obtains a hybridization result based on the information about the state of the biochip acquired by the imaging device.

[0009] Optionally, multiple test tube vials are used, the cavity is divided into several spaces for carrying different test tube vials, and the top surface of each of the spaces is configured as an insertion opening for inserting a corresponding test tube vial.

[0010] Optionally, a hollowed portion is provided on one side of the cavity, and the imaging device acquires the information about the biochip in the bottle body of the test tube vial through the hollowed portion; and the bottle body of the test tube vial is a flat structure, and / or the bottle body is made of a translucent material.

[0011] Optionally, the cavity is arranged to conform to the structure of the bottle body of the test tube vial; and / or the upper part of the cavity is provided with a recess for receiving the bottle cap of the test tube vial.

[0012] Optionally, the biochip detection device further comprises a holder configured to support the liquid separation needle, which is arranged above the cavity and is movable upwards and downwards; and the holder is provided with a liquid separation needle hole for the passage of the liquid separation needle.

[0013] Optionally, the bottle cap of the test tube vial is arranged corresponding to the liquid separation needle; optionally, the liquid separation needle is arranged inserted into the bottle cap of the corresponding test tube vial and is inserted into the test tube vial.

[0014] Optionally, a drainage needle is also provided, which is configured to drain the liquid in the test tube vial and which is in fluid communication with a drainage device.

[0015] Optionally, the bracket is configured to support the drainage needle, and optionally, the bracket is provided with a drainage needle hole for the passage of the drainage needle.

[0016] Optionally, the drainage needle and the liquid separation needle are arranged adjacent to each other, and one liquid separation needle and one drainage needle are provided corresponding to the bottle cap of the test tube vial.

[0017] Optionally, both ends of the cavity are each provided with a guide rail along the vertical direction, and both ends of the bracket are each slidably connected to a corresponding guide rail by means of a guide block.

[0018] Optionally, the bottom of the cavity is provided with a heating and cooling plate, and the temperature control of the cavity is realized by cooling and heating conduction.

[0019] Optionally, the heating and cooling plate is a thermoelectric cooling plate in which the conversion between the heating function and the cooling function is realized by a control electrode.

[0020] Optionally, a heat dissipation device is arranged under the heating and cooling plate, e.g. at least one from the group consisting of a cooling fin and a fan.

[0021] Optionally, multiple test tube vials are used, and the cavity is divided into spaces for carrying different test tube vials.

[0022] Optionally, the liquid storage device comprises a plurality of liquid storage bottles, and the plurality of liquid storage bottles control a mixed discharge of different solutions through a liquid separation valve.

[0023] Optionally, the fluid isolation valve comprises one of the group consisting of an eight-way valve, a four-way valve, and a three-way valve, or a combination of several thereof.

[0024] Optionally, a cover plate for securing the test tube vial is arranged between the holder and the cavity; and optionally, the cover plate is provided with holes corresponding to the liquid separation needle and the drainage needle, respectively.

[0025] Optionally, a platform is included, with the cavity, fluid storage device, pumping device, holder, and imaging device all mounted on the platform.

[0026] Optionally, the cavity is mounted on one side edge of the platform, the bracket is mounted above the cavity, a mounting plate is fixed vertically on the other side edge of the platform facing the cavity, the liquid storage bottle of the liquid storage device is fixed on a side surface of the mounting plate facing away from the cavity, the pumping device is located between the cavity and the mounting plate, and the imaging device is mounted in parallel on one side of the cavity.

[0027] Optionally, the imaging device is arranged to be movable.

[0028] Optionally, the hollowed side of the cavity is provided with a rail along a direction parallel to the cavity, and the imaging device is slidably connected to the rail by a carriage.

[0029] The present disclosure further provides a biochip detection method performed using a biochip detection apparatus as described above, comprising the steps of: Inserting a biochip, a sample to be detected, and a primer into the test tube vial fixed in the cavity; Carrying out the amplification of the sample to be detected and the hybridization of the amplification product with the biochip while controlling the inflow and outflow of the liquid in the test tube vial and controlling the temperature; and Performing appropriate treatment using various detection methods, acquiring the information about the fully hybridized biochip by an imaging device, and processing the hybridization result by a data processing device to obtain a result.

[0030] A pumping device optionally pumps the liquid in a liquid storage device through a liquid separation needle into the test tube vial, and the liquid in the test tube vial is pumped through a drainage needle into a drainage device.

[0031] Optionally, the amplification of the sample to be detected and the hybridization of the amplification product with the biochip are carried out simultaneously, and the buffer solution used is a PCR amplification buffer solution.

[0032] A temperature control program can be adapted depending on the object to be detected and the detection method. For example, it could be a typical PCR reaction program, or a dual-temperature PCR, such as a dual-temperature PCR of 94°C and 55°C, or a constant-temperature amplification such as LAMP / RPA.

[0033] Optionally, the temperature control program consists of several cycles of denaturation, annealing, and stretching.

[0034] Optionally, the temperature control program has 35-40 cycles of 94 ± 1°C for 10 ± 2 s, 55 ± 1°C for 20 ± 2 s, and 72 ± 2°C for 20 ± 2 s.

[0035] Optionally, the detection method includes a chemiluminescence method, a fluorescence method, and a visualization method.

[0036] During practical operations, different detection methods can be used in detection, and the detection could also be performed using the same detection method.

[0037] If different detection methods are used, a reagent affected by the corresponding detection method is added each time, an image is displayed with a corresponding procedure, and the process is repeated several times to obtain hybridization results of different objects to be detected of the corresponding sample.

[0038] When using the same detection method, different markers can be selected depending on the different objects to be detected and treated under different conditions to obtain hybridization results for different objects to be detected in the corresponding sample. For example, in the chemiluminescence method, a horseradish enzyme-labeled avidin solution is added; after hybridization is complete, a luminescent substrate is added; and a high-sensitivity CCD imaging element (model JAI CM-030-GE) moves along the guide rail to just in front of the respective chips, capturing a chemiluminescent biochip image.

[0039] For example, in the fluorescence method, an avidin-labeled red quantum dot SA-QDs 625 solution is added; after hybridization is completed, the ultraviolet light source is excited, followed by filtration; and a highly sensitive CCD imaging element moves on the guide rail to just in front of each chip, capturing a quantum dot fluorescence biochip image.

[0040] For example, in the visualization process, an avidin-labeled red quantum dot SA-QDs 625 solution is added; after hybridization is completed, a silver-stained substrate solution is added, followed by cleaning; and a white LED light source is turned on, and the CCD imaging element moves on the guide rail to just in front of respective chips, capturing a visualization biochip image.

[0041] In addition to quantum dots and biotin as described above, the substances concerned in the present disclosure and used to display signals may also include: a fluorescent substance, a digoxin-labeled probe, a radioactive isotope, a radioactive contrast agent, a fluorescent microsphere for paramagnetic ions, an electron-dense substance, a chemiluminescent marker, an ultrasound contrast agent, a photosensitizer, colloidal gold, or enzyme.

[0042] The fluorescent substance includes any of Alexa 350, Alexa 405, Alexa 430, Alexa 488, Alexa 555, Alexa 647, AMCA, Aminoacridine, BODIPY 630 / 650, BODIPY 650 / 665, BODIPY-FL, BODIPY-R6G, BODIPY-TMR, BODIPY-TRX, 5-carboxy-4', 5'-dichloro-2', 7'-dimethoxyfluorescein, 5-carboxy-2', 4', 5', 7'-tetrachlorofluorescein, 5-carboxyfluorescein, 5-carboxyrhodamine, 6-carboxytetramethylrhodamine, Cascade Blue, Cy2, Cy3, Cy7, 6-FAM, Dansyl Chloride, Fluorescein, HEX, 6-JOE, NBD (7-Nitrobenzo-2-oxa-1,3-diazole), Oregon Green 488, Oregon Green 500, Oregon Green 514, Pacific Blue, Phthalic Acid, Terephthalic Acid, Isophthalic Acid, Cresol Fast Violet, Cresol Blue Violet, Brilliant Cresol Blue, p-Aminobenzoic Acid, Erythrosine, Phthalocyanine, Azomethine, Cyanine, Xanthine, Succinylfluorescein, Rare Earth Metal Cryptate, Tripyridyldiamine Europium, Europium Cryptate or Chelate, Diamine, Bispyanine Glycoside, La Jolla Blue Dye, Allophycocyanin, Allophycocyanin B, Phycocyanin C, Phycocyanin R, Thiamin,Phycoerythrin R, REG, rhodamine green, rhodamin isothiocyanate, rhodamine red, ROX, TAMRA, TET, TRIT (tetramethyl rhodamine isothiol), tetramethyl rhodamine or Texas red.

[0043] The radioactive isotope includes any of 110 In, 111 In, 177 Lu, 18 F, 52 Fe, 62 Cu, 64 Cu, 67 Cu, 67 Ga, 68 Ga, 86 Y, 90 Y, 89 Zr, 94 mTc, 94 Tc, 99 mTc, 120 I, 123 I, 124 I, 125 I 13 I, 154-158 Gd, 32 P, 11 C, 13 N, 15 O, 186 Re, 188 Re, 51 Mn, 52 In my opinion, 55 Co, 72 Ace, 75 Br, 76 Br, 82 mRb or 83 Sr.

[0044] The enzyme includes any of horseradish peroxidase, alkaline phosphatase, or glucose oxidase.

[0045] The fluorescent microspheres are: fluorescent polystyrene microspheres wrapped inside with the fluorescent rare earth ion europium.

[0046] Compared to the prior art, the present disclosure has at least the following advantageous effects: (1) In this disclosure, a cavity having a temperature control function is provided so that the amplification of a sample to be detected and the hybridization of a biochip are operated in an integrated manner, thereby greatly reducing the operation steps and saving time. (2) In this disclosure, the mixing of different liquids is controlled by the liquid separation valve, and the mixed liquid flows into a test tube vial through the liquid separation needle to meet the requirement of solutions at different stages, at the same time, the waste liquid is discharged through the drainage needle, thereby realizing full automation of the hybridization process. (3) In this disclosure, an imaging device is still provided to acquire information about biochips in bottle bodies, thereby automating the acquisition of results and saving labor. (4) In the biochip detection device provided in this disclosure, the entire process of biochip detection is automated, and high detection efficiency and stable and reliable detection results are achieved. Description of the drawings

[0047] In order to more clearly describe technical solutions in specific embodiments of the present disclosure or in the prior art, the drawings necessary in explaining the specific embodiments or the prior art are briefly presented below; and of course, the drawings in the following explanation show only some embodiments of the present disclosure, and other drawings may be available to those skilled in the art by referring to these drawings without exercising an inventive step. Fig. 1 shows a front perspective view of a biochip detection device according to an embodiment of the present disclosure; Fig. 2 shows a perspective side view of the biochip detection device according to an embodiment of the present disclosure; Fig. 3 shows a rear perspective view of the biochip detection device according to an embodiment of the present disclosure; Fig. 4 shows a schematic structural diagram of a cavity in the biochip detection device according to an embodiment of the present disclosure; Fig. 5 shows a schematic structural diagram of a holder in the biochip detection device according to an embodiment of the present disclosure; Fig. 6 shows a dot array diagram of various samples of a biochip according to an embodiment of the present disclosure; Fig. 7 shows a diagram of the chemiluminescence detection result of the biochip according to an embodiment of the present disclosure; Fig. 8 shows a diagram of the fluorescence detection result of the biochip according to an embodiment of the present disclosure; and Fig. 9 shows a diagram of the visualization detection result of the biochip according to an embodiment of the present disclosure. Reference symbol:

[0048] 1-cavity; 2-liquid storage bottle; 3-pumping device; 4-holder; 5-imaging device; 6-liquid separation valve; 7-space; 8-insertion port; 9-hollowed section; 10-recess; 11-liquid separation needle hole; 12-drainage needle hole; 13-guide block; 14-guide rail; 15-platform; 16-mounting plate; and 17-printed circuit board. Detailed description of the embodiments

[0049] In the following, the technical solutions of this disclosure will be described clearly and comprehensively with reference to the drawings and specific embodiments. However, it should be understood by those skilled in the art that the embodiments described below are only partial embodiments, rather than all of the embodiments of the disclosure, serve only to describe the present disclosure, and should not be regarded as limitations on the scope of this disclosure. Any other embodiments that could be obtained by those skilled in the art based on the embodiments in this disclosure without inventive step fall within the scope of the present disclosure. Examples for which no specific conditions are written are carried out under conventional conditions or the conditions recommended by the manufacturer.Reagents or instruments used for which no manufacturer is specified are conventional products that are commercially available.

[0050] In explaining this disclosure, it should be explained that orientation or positional relationships indicated by terms such as "central," "top," "bottom," "left," "right," "vertical," "horizontal," "inside," and "outside" are orientation or positional relationships shown based on the drawings. These terms are intended only to facilitate explanation of the disclosure and simplify explanation, rather than to imply or imply that the mentioned device or element should have a specific orientation and be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on the disclosure. Furthermore, terms such as "first," "second," and "third" are for illustrative purposes only and should not be interpreted as implying or implying any importance in relativity.

[0051] In explaining this disclosure, it should be clarified that terms such as "assemble," "connection," and "connect" should be understood in a broad sense unless otherwise expressly stated and defined. For example, it could be either a fixed connection, a detachable connection, or an integrated connection; it could be either a mechanical connection or an electrical connection; and it could be either a direct connection, an indirect connection through an intermediate link, or an internal communication between two elements. Those skilled in the art could understand the specific meanings of the above terms in this disclosure according to specific circumstances.

[0052] As it is in Fig. 1-3, a biochip detection device provided in this disclosure comprises a cavity 1 for supporting a test tube vial, the test tube vial for containing a sample to be detected and a biochip, and the cavity 1 has a temperature control function; a liquid storage device for storing a liquid; a pumping device 3 for pumping the liquid in the liquid storage device into the test tube vial, the pumping device 3 being in fluid communication with a liquid separation needle, the liquid separation needle being configured to inject a mixed liquid into the test tube vial; an imaging device 5 for monitoring the state of a chip in the test tube vial; and a data processing device that obtains a hybridization result based on the information about the state of the biochip acquired by the imaging device 5.

[0053] It should be explained that the existing procedure for biochip hybridization and detection of hybridization results is carried out as follows: Preparing a biochip with different probes: Different probe buffer solutions are spotted onto a substrate using a gene chip sample applicator and dried for later use; Preparing a PCR product for hybridization: A PCR amplification is performed on an object to be detected; and Performing hybridization on the biochip and the PCR product: The PCR amplification product is denatured (98°C), immediately subjected to a cold bath, and then mixed with a hybridization buffer solution, the mixed liquid is added to a sample application area of ​​the chip, the chip is placed in a hybridization box and subjected to hybridization in an oven in a light-tight state; and after washing in a light-tight state, the result is scanned and analyzed.

[0054] In the biochip detection device provided in the present disclosure, a sample to be detected and a biochip are directly placed in a test tube vial; the temperature is controlled by the temperature control function of the cavity 1; an appropriate solution in the liquid storage device flows into the test tube vial under the action of the pumping device 3, or the discharge of the liquid in the test tube vial is controlled; the sample to be detected is hybridized with the biochip while undergoing PCR amplification; then, treatment is performed by adding an appropriate washing liquid using various detection methods; the imaging device 5 acquires the information about the hybridized biochip, and based on the hybridization result, a result is obtained after processing by the data processing device.

[0055] It should be noted that the biochip according to the present disclosure is not only a gene chip but also a protein chip. In other words, the biochip detection device according to the present disclosure can be applied to both genetic detection and protein detection, such as immunodetection.

[0056] The present disclosure automates the entire biochip detection process, and high detection efficiency and stable and reliable detection results are achieved.

[0057] In the present disclosure, the cavity 1 serves to support a test tube vial, and the cavity 1 may be completely surrounded by a side wall or consist of a frame. To realize the analysis of multiple biological samples at once, multiple test tube vials may be used; with reference to Fig. 4, the cavity 1 is divided into a plurality of spaces 7 for supporting different test tube vials, the top surface of each of the spaces 7 is configured as an insertion opening 8 for inserting a corresponding test tube vial, in the cavity 1 a plurality of individual spaces 7 are provided, and each of the spaces 7 serves to place only one test tube vial.

[0058] To facilitate image taking and temperature control, a hollowed section 9 ( Fig. 4) in some embodiments is provided on one side of the cavity 1, and the imaging device 5 acquires the information about the biochip in the bottle body through the hollowed portion 9.

[0059] Optionally, the camera of the imaging device 5 is directed directly at the hollowed section 9 to facilitate image capture.

[0060] In some embodiments, the bottle body of the test tube vial is a flat structure. The bottle body serves to accommodate the biochip and the sample to be detected. However, the biochip is generally in the shape of a quadrangular sheet, while the sample to be detected is a liquid. For this reason, the shape of the bottle body is adapted to the biochip; and the volume shape of the bottle body is slightly larger than that of the biochip. The flat structure not only facilitates easy insertion of the biochip, but the orientation of the plane of the flat structure toward the imaging device 5 also facilitates the acquisition of information about the biochip by the imaging device 5.

[0061] In some embodiments, the bottle body is made of a translucent material. This ensures the clarity of the image in the bottle body, facilitating the acquisition of information about the biochip by the imaging device 5.

[0062] In some embodiments, the cavity 1 is arranged to adapt to the structure of the bottle body; specifically, the size of each space 7 is adjusted to adapt to the size of the corresponding test tube vial. Since the cavity 1 has a temperature control property, but the temperature of the test tube vial is often controlled by the heat conduction of the cavity 1, the cavity 1 is provided to adapt to the structure of the bottle body, thereby also improving the stability when placing the test tube vial. Optionally, the bottle body is a flat structure as an example, see Fig. 4, and the shape for supporting a test tube vial is a square, flat shape, which not only facilitates heat conduction but also allows the test tube vial to be securely inserted into a corresponding space 7. In some embodiments, the top of the cavity 1 is provided with a recess 10 for receiving the bottle cap of the test tube vial. The provision of the recess 10 positions the bottle cap of the test tube vial, and the stability of the bottle cap is also enhanced, thereby facilitating the insertion of the liquid separation needle or another infusion needle into or withdrawal from the bottle cap.

[0063] In some embodiments, the biochip detection device further comprises a holder 4 for supporting the liquid separation needle, wherein the holder 4 is arranged above the cavity 1 and is movable upwards or downwards.

[0064] The liquid separation needle is arranged above the test tube vial and is located on the holder 4, wherein the insertion of the liquid separation needle into the test tube vial or its withdrawal from it is realized by the movement of the holder 4. As an example, the holder 4 is provided with a liquid separation needle hole 11, which serves for the passage of the liquid separation needle, see Fig. 5; and the liquid separation needle is arranged to be inserted into the liquid separation needle hole 11, thereby realizing a function of supporting the liquid separation needle.

[0065] In some embodiments, the bottle cap of the test tube vial is arranged correspondingly to the liquid separation needle, in other words, only one test tube vial is carried in a space 7, a liquid separation needle is arranged correspondingly inserted into the bottle cap of a test tube vial; optionally, the liquid separation needle is arranged inserted into the bottle cap of the corresponding test tube vial and is inserted into the test tube vial.

[0066] Different treatment solutions are required at different stages of the hybridization process, and automated fluid exchange operations can be realized by flowing in and out of the fluid.

[0067] In some embodiments, a drainage needle is also provided, which is configured to drain the liquid in the test tube vial and is in fluid communication with a drainage device. The drainage needle drains the liquid into the drainage device, such as a waste bottle.

[0068] In some embodiments, the holder 4 is further configured to support the drainage needle, and optionally the holder is provided with a drainage needle hole 12 which serves for the passage of the drainage needle, see Fig. 5; and the drainage needle is arranged to be inserted into the drainage needle hole 12, thereby realizing a function of supporting the drainage needle.

[0069] In some embodiments, the drainage needle hole 12 and the liquid separation needle hole 11 are provided in pairs, see Fig. 5; The drainage needle hole 12 and the liquid separation needle hole 11, which are provided in pairs, are arranged adjacent to each other. Accordingly, the drainage needle and the liquid separation needle are arranged adjacent to each other. A liquid separation needle and a drainage needle are provided corresponding to the bottle cap of the test tube vial. In other words, automated liquid exchange operations are realized in each test tube vial by controlling the inflow of a liquid through the liquid separation needle and controlling the discharge of the liquid through the drainage needle.

[0070] In some embodiments, both ends of the cavity 1 are each provided with a guide rail 14 along the vertical direction, and both ends of the holder 4 are each slidably connected to a corresponding guide rail 14 by means of a guide block 13. By controlling the movement of the holder 4 along the guide rail 14, an upward and downward movement of the holder 4 is realized, whereby the liquid separation needle and the dewatering needle are driven to move upward and downward, respectively, and are therefore inserted into and withdrawn from the bottle cap of the test tube vial.

[0071] In some embodiments, the bottom of the cavity 1 is provided with a heating and cooling plate, and the temperature control of the cavity 1 is realized by cooling and heating conduction.

[0072] In some embodiments, the heating and cooling plate is a thermoelectric cold plate in which the conversion between the heating function and the cooling function is realized by a control electrode.

[0073] In some embodiments, a heat dissipation device is arranged below the heating and cooling plate, and optionally the heat dissipation device comprises at least one of the group consisting of a cooling fin and a fan.

[0074] A controlled temperature in the test tube vial is realized by the heating device and the heat dissipation device.

[0075] In some embodiments, the liquid storage device comprises a plurality of liquid storage bottles 2, and the plurality of liquid storage bottles 2 control a mixed discharge of different solutions through a liquid separation valve 6. The liquid storage bottle 2 is in fluid communication with the liquid separation needle, and specifically, the liquid storage bottle 2 is in fluid communication with the pumping device 3, and the pumping device 3 is in fluid communication with the liquid separation needle.

[0076] In some embodiments, the liquid separation valve 6 comprises one of the group consisting of an eight-way valve, a four-way valve, and a three-way valve, or a combination of several thereof.

[0077] In some embodiments, a cover plate for securing the test tube vial is further arranged between the holder 4 and the cavity 1, and the cover plate is further provided with holes corresponding respectively to the liquid separation needle and the drainage needle.

[0078] By providing the cover plate, the test tube vial is placed more firmly.

[0079] In some embodiments, a platform 15 is also included, with the cavity 1, the fluid storage device, the pumping device 3, the holder 4, and the imaging device 5 all mounted on the platform 15, with all units being integrated as a whole by the platform 15. Furthermore, the cover plate is elastically connected to the platform 15.

[0080] In some embodiments, the cavity 1 is mounted on one side edge of the platform 15, the holder 4 is mounted above the cavity 1, a mounting plate 16 is fixed vertically on the other side edge of the platform 15 facing the cavity, the liquid storage bottle 2 of the liquid storage device is fixed to a side surface of the mounting plate 16 facing away from the cavity, the pumping device 3 is located between the cavity 1 and the mounting plate 16, the imaging device 5 is mounted in parallel on one side of the cavity 1, and a vertical arrangement of multiple liquid storage bottles 2 is realized by the mounting plate 16. A printed circuit board 17 is mounted vertically on the side edge of the platform 15 opposite the cavity 1, and the liquid separation valve is located between the printed circuit board 17 and the cavity 1.

[0081] In some embodiments, the imaging device 5 is arranged to be movable.

[0082] In some embodiments, the side of the cavity 1 provided with a hollowed portion 9 is provided with a rail along a direction parallel to the cavity 1, and the imaging device 5 is slidably connected to the rail by a carriage.

[0083] During practical operations, the position of the imaging device 5 is controlled at different locations depending on the test tube vials to enable the acquisition of information about biochips in different test tube vials.

[0084] In the biochip detection apparatus provided in the present disclosure, the flow of the liquid and the movement of the holder 4, etc., are completed by a control system, and the control system and a data processing system are provided on the same operation side.

[0085] The control system includes a printed circuit board 17 and the like, for example, a microprocessor chip, a temperature control module, a drive module, a touch liquid crystal display module, a USB module, a JTAG module, an alarm module and the like are mounted on the printed circuit board 17, and the microprocessor chip communicates with respective modules and controls the respective modules to realize accurate execution of the entire operations.

[0086] The present disclosure further provides a biochip detection method performed using a biochip detection apparatus as described above, comprising the steps of: Placing a biochip, a sample to be detected, and a primer in a test tube vial fixed in the cavity 1; Carrying out the amplification of the sample to be detected and the hybridization of the amplification product with the biochip while controlling the inflow and outflow of the liquid in the test tube vial and controlling the temperature; and Performing appropriate treatment using various detection methods, acquiring the information about the fully hybridized biochip by an imaging device 5, and processing the hybridization result by a data processing device to obtain a result.

[0087] In some embodiments, the pumping device 3 pumps the liquid in the liquid storage bottle 2 through the liquid separation valve 6 and the liquid separation needle into the test tube vial, and the liquid in the test tube vial is pumped through the drainage needle into a drainage device, thereby realizing the inflow and outflow of the liquid in the test tube vial.

[0088] In some embodiments, the information about the biochip in this test tube vial is acquired by sliding the imaging device 5 on the rail until it is directly aimed at the hollowed portion 9 of the space 7 where the target test tube vial is located.

[0089] It should be explained that schematic diagrams of the biochip detection device provided in this disclosure are shown without various piping assembled, and in use, appropriate assembly is required before use.

[0090] The following examples are specifically listed: Influenza A, influenza B, RSV, and adenovirus from samples were detected, and primers and probes of these multiple objects to be detected were designed according to a conventional method, specifically as shown in Table 1.

[0091] Nucleic acid probe sample application: The biochip was a self-preparable aldehyde slide, the concentration of probe sample applications was 10 µm, and the samples were stored in a dry state at room temperature, the schematic representation of which is shown in Fig. 6 is shown.

[0092] PCR amplification: A nucleic acid sample (the object to be detected), a biochip, and a PCR mix reagent (including primers and buffer solution) were mixed and added to a test tube vial of the instrument. The thermal cycling parameters of the instrument's semiconductor heating and cooling plate were as follows: 94°C for 10 s, 55°C for 20 s, and 72°C for 20 s, for 40 cycles, achieving simultaneous amplification and hybridization. A washing fluid was then pumped in through an injection pump and a solenoid valve to clean the chip.

[0093] Subsequently, three detection methods, namely chemiluminescence, quantum dot fluorescence and visualization, were realized, each with three types of instruments and reagents.

[0094] Detection by the chemiluminescence method: 100 µL of the horseradish enzyme-labeled avidin solution was automatically pumped into each reaction bottle and reacted for 30 min at 37°C. A PBST wash solution was pumped in for cleaning. Then, 100 µL of chemiluminescent substrate solution (the ratio of liquid A to liquid B in Millipore hypersensitive chemiluminescent substrate solution was 1:1) was pumped in. A high-sensitivity CCD imaging element (model JAI CM-030-GE) moved along the guide rail to just in front of each chip, capturing a chemiluminescent biochip image. The exposure time for each chip was 10 s, and a total of 8 chips were detected. The results are shown in Fig. 7. This image result indicates that this sample is positive for influenza A nucleic acid.

[0095] Detection by fluorescence method: 100 µL of 25 nM avidin-labeled red quantum dot SA-QDS 625 solution (Wuhan Jiayuan) was automatically pumped into each reaction bottle and reacted for 30 min at 37°C. A PBST wash solution was pumped in for cleaning. A 5 W UV LED light source with a wavelength of 365 nm was switched on to excite the QDs and emit fluorescence. The fluorescence was filtered through a filter (624 / 40 nm, model Edmund Optical 67-021). The highly sensitive CCD imaging element moved along the guide rail to just in front of each chip, capturing a quantum dot fluorescence biochip image. The exposure time for each chip was 10 s, and a total of 8 chips were detected. The results are shown in Fig. 8. This image result indicates that this sample is positive for influenza A nucleic acid.

[0096] Detection by the visualization method: 100 µL of 25 nM avidin-labeled red quantum dot SA-QDS 625 solution (Wuhan Jiayuan) was automatically pumped into each reaction bottle and reacted for 30 min at 37°C. A PBST wash solution was pumped in for cleaning. Then, 100 µL of silver staining substrate solution was pumped in, followed by a reaction for 5 minutes, and the product was washed with deionized water. A white LED light source was switched on, and a high-sensitivity CCD imaging element moved along the guide rail to just in front of each chip, capturing a visualization biochip image, and a total of 8 chips were detected. The results are shown in Fig. 9. This image result indicates that this sample is positive for influenza A nucleic acid.

[0097] In the biochip detection device provided in this disclosure, a sample to be detected and a biochip are directly added to a test tube vial; by controlling the temperature and the inflow and outflow of an appropriate solution, the reaction product of PCR amplification of the sample during amplification with the biochip is detected; then, an image of the chip is captured after adding an appropriate washing liquid to obtain information, whereby the final hybridization result can be obtained. The integration of the hybridization process of a gene chip and the output of results is realized, the processing efficiency of samples is increased, labor is saved, and stable and reliable results can be obtained.

[0098] Finally, it should be explained that the above respective embodiments serve only to describe the technical solutions of the present disclosure, rather than limiting them. Although the present disclosure is described in detail with reference to the foregoing respective embodiments, those skilled in the art should understand that the technical solutions recorded in the foregoing respective embodiments may still be modified, or partial or all technical features may be substituted therein, while these modifications or substitutions do not cause the essence of the respective technical solutions to deviate from the scope of the technical solutions of the respective embodiments of this disclosure. Industrial applicability

[0099] The present disclosure provides a biochip detection apparatus and a biochip detection method, wherein the integration of the reaction of a biochip and the output of detection results is realized, the entire detection process is automated, the treatment efficiency of samples is increased, labor is saved, and stable and reliable results can be obtained.

Claims

[1] Detection device for biochips, characterized by that the detection device for biochips comprises the following components: a cavity (1) for carrying a test tube vial, wherein the test tube vial serves to hold a sample to be detected and a biochip, and the cavity (1) has a temperature control function; a liquid storage device for storing a liquid; a pumping device (3) for pumping the liquid in the liquid storage device into the test tube vial, the pumping device (3) being in fluid communication with a liquid separation needle, the liquid separation needle being configured to inject a mixed liquid into the test tube vial; an imaging device (5) for monitoring the state of a chip in the test tube vial; and a data processing device that obtains a hybridization result based on the information about the state of the biochip acquired by the imaging device (5). [2] Biochip detection device according to claim 1, characterized by that several test tube bottles are used, the cavity (1) is divided into several spaces for carrying different test tube bottles, and the cover surface of each of the spaces is configured as an insertion opening (8) for inserting a corresponding test tube bottle. [3] Biochip detection device according to claim 1, characterized bythat a hollowed portion (9) is provided on one side of the cavity (1), and the imaging device (5) acquires the information about the biochip in the bottle body of the test tube vial through the hollowed portion (9); and the bottle body of the test tube vial is a flat structure, and / or the bottle body is made of a translucent material. [4] Detection device for biochips according to one of claims 1-3, characterized by that the cavity (1) is arranged such that it adapts to the structure of the bottle body of the test tube bottle; and / or the upper part of the cavity (1) is provided with a recess (10) for receiving the bottle closure of the test tube bottle. [5] Detection device for biochips according to one of claims 1-4, characterized bythat a holder (4) is also included, which is configured to support the liquid separation needle and which is arranged above the cavity (1) and is movable upwards and downwards; and the holder (4) is provided with a liquid separation needle hole (11) which serves for the passage of the liquid separation needle. [6] Biochip detection device according to claim 5, characterized by that the bottle cap of the test tube vial is arranged corresponding to the liquid separation needle; optionally, the liquid separation needle is arranged inserted on the bottle cap of the corresponding test tube vial and is inserted into the test tube vial. [7] Detection device for biochips according to claim 5 or 6, characterized bythat a drainage needle is also provided which is configured to drain the liquid in the test tube vial and which is in fluid communication with a drainage device. [8] Detection device for biochips according to claim 7, characterized by that the holder (4) is configured to support the drainage needle, and optionally the holder (4) is provided with a drainage needle hole (12) which serves for the passage of the drainage needle. [9] Detection device for biochips according to claim 7 or 8, characterized by that the drainage needle and the liquid separation needle are arranged adjacent to each other, and a liquid separation needle and a drainage needle are provided corresponding to the bottle cap of the test tube vial. [10] Detection device for biochips according to one of claims 5-9, characterized bythat both ends of the cavity (1) are each provided with a guide rail (14) along the vertical direction, and both ends of the holder (4) are each slidably connected to a corresponding guide rail (14) by means of a guide block (13). [11] Biochip detection device according to claim 1, characterized by that the bottom of the cavity (1) is provided with a heating and cooling plate, and the temperature control of the cavity (1) is realized by cooling and heat conduction; optionally the heating and cooling plate is a thermoelectric cooling plate in which the conversion between the heating function and the cooling function is realized by a control electrode. [12] Biochip detection device according to claim 11, characterized bythat a heat dissipation device is arranged below the heating and cooling plate; optionally the heat dissipation device comprises at least one from the group consisting of a cooling fin and a fan. [13] Biochip detection device according to claim 1, characterized by in that the liquid storage device comprises a plurality of liquid storage bottles (2), and the plurality of liquid storage bottles (2) control a mixed discharge of different solutions through a liquid separation valve (6); and optionally the liquid separation valve (6) comprises one of the group consisting of an eight-way valve, a four-way valve, and a three-way valve, or a combination of several thereof. [14] Detection device for biochips according to one of claims 8-10, characterized bythat a cover plate for fastening the test tube vial is arranged between the holder (4) and the cavity (1); and optionally the cover plate is provided with holes corresponding respectively to the liquid separation needle and the drainage needle. [15] Biochip detection device according to claim 1, characterized by that a platform (15) is still included, wherein the cavity (1), the liquid storage device, the pumping device (3), the holder (4), and the imaging device (5) are all mounted on the platform (15). [16] Biochip detection device according to claim 15, characterized bythat the cavity (1) is mounted on a side edge of the platform (15), the holder (4) is mounted above the cavity (1), a mounting plate (16) is fixed vertically on the other side edge of the platform (15) facing the cavity (1), the liquid storage bottle (2) of the liquid storage device is fixed on a side surface of the mounting plate (16) facing away from the cavity (1), the pump device (3) is located between the cavity (1) and the mounting plate (16), and the imaging device (5) is mounted in parallel on one side of the cavity (1). [17] Detection device for biochips according to one of claims 1-16, characterized by that the imaging device (5) is arranged to be movable. [18] Biochip detection device according to claim 17, characterized bythat the hollowed side of the cavity (1) is provided with a rail along a direction parallel to the cavity (1), and the imaging device (5) is slidably connected to the rail by a carriage. [19] Detection methods for biochips, characterized by that the biochip detection method is carried out using a biochip detection device according to any one of claims 1-18 and comprises the following steps: Inserting a biochip, a sample to be detected, and a primer into the test tube vial fixed in the cavity (1); Carrying out the amplification of the sample to be detected and the hybridization of the amplification product with the biochip while controlling the inflow and outflow of the liquid in the test tube vial and controlling the temperature; and Performing appropriate treatment using various detection methods, acquiring the information about the fully hybridized biochip by an imaging device (5), and processing the hybridization result by a data processing device to obtain a result; wherein the biochip is a gene chip, the amplification of the sample to be detected and the hybridization of the amplification product with the biochip are carried out simultaneously, and the buffer solution used is a PCR amplification buffer solution; wherein a temperature control program consists of several cycles of denaturation, annealing, and stretching; wherein the temperature control program further comprises 35-40 cycles of 94 ± 1°C for 10 ± 2 s, 55 ± 1°C for 20 ± 2 s, and 72 ± 2°C for 20 ± 2 s; and wherein the detection method further comprises a chemiluminescence method, a fluorescence method, and a visualization method. [20] Detection method for biochips according to claim 19, characterized by that by a pumping device (3), the liquid in a liquid storage device is pumped through a liquid separation needle into the test tube vial, and the liquid in the test tube vial is pumped through a drainage needle into a drainage device.

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