CN109975564B - Automatic detection device for microfluid immunoassay integrated reagent - Google Patents
Automatic detection device for microfluid immunoassay integrated reagent Download PDFInfo
- Publication number
- CN109975564B CN109975564B CN201910217642.XA CN201910217642A CN109975564B CN 109975564 B CN109975564 B CN 109975564B CN 201910217642 A CN201910217642 A CN 201910217642A CN 109975564 B CN109975564 B CN 109975564B
- Authority
- CN
- China
- Prior art keywords
- sample
- liquid
- substrate liquid
- tube
- hematocrit
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54306—Solid-phase reaction mechanisms
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/544—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being organic
- G01N33/545—Synthetic resin
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
- G01N35/00029—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor provided with flat sample substrates, e.g. slides
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
- G01N2035/00178—Special arrangements of analysers
- G01N2035/00237—Handling microquantities of analyte, e.g. microvalves, capillary networks
Landscapes
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Physics & Mathematics (AREA)
- Pathology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Plasma & Fusion (AREA)
- Optics & Photonics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention relates to an automatic detection device for a microfluid immunoassay integrated reagent, which can comprise a working turntable, an automatic sample introduction device, a luminous substrate liquid filling device, a hematocrit scale reader and a fluorescence/luminescence detection unit, wherein the working turntable is divided into even number of reagent positions, each reagent position can be fixedly provided with the microfluid immunoassay integrated reagent device, the working turntable is symmetrically provided with a plurality of hematocrit test tube jigs, the hematocrit test tubes are arranged in the hematocrit test tube jigs, the automatic sample introduction device is used for adding a blood sample to be detected into the microfluid immunoassay integrated reagent device, the luminous substrate liquid filling device is used for filling luminous substrate liquid into a characteristic marker reaction tube, the hematocrit scale reader is used for reading the hematocrit number in the centrifuged hematocrit test tube, and the fluorescence/luminescence detection unit is used for carrying out fluorescence/luminescence detection on the sample after reaction and washing in the microfluid immunoassay integrated reagent device.
Description
Technical Field
The invention relates to a structural device for automatically detecting a servo instant detection integrated reagent tube, in particular to a microfluid immunoassay integrated reagent automatic detection device for realizing high-sensitivity instant detection in medical diagnosis, health monitoring, food safety and other emergency places.
Background
In medical diagnosis, health monitoring, food safety and other emergency places, automatic high-sensitivity instant detection always has a real and vigorous demand.
Currently, the labeling immune instant detection based on lateral chromatography is widely applied, and labels relate to colloidal gold, fluorescence, lanthanide rare earth element Eu2+, quantum dots and the like. The method has the greatest advantages of simple and convenient detection operation, short instrument structure and capability of meeting the requirement of instant detection in a plurality of places. The labeled immunoreagent material based on lateral chromatography is poor in quality controllability, large in uncertainty of production process control, unstable in property of reagent assembly materials and uncontrollable in detection process, detection results are low in sensitivity and large in variation, and clinical application is prone to occurrence of conditions such as missed diagnosis and misdiagnosis, so that the labeled immunoreagent material is only used for screening detection of high risk groups.
Other forms of real-time detection including magnetic particle solid-phase chemiluminescence immunoassay, real-time detection in microfluidic drive mode, etc. are also under development and application at the same time. However, the simplicity and convenience of analysis are more complicated than those of labeled immune real-time detection of lateral chromatography, and the composition of reagents and instruments requires more operation and auxiliary conditions. There is no doubt that more sensitive and simple methods like lateral chromatographic labeled immunodetection are desired to meet the needs of the real world.
Disclosure of Invention
The invention aims to provide an automatic detection device for a microfluid immunoassay integrated reagent, which utilizes capillary action and centrifugal force to drive the reaction process of the microfluid immunoassay integrated reagent device, completes detection and reports the result qualitatively and quantitatively. Particularly, the automatic washing of the detection tube can be realized, so that the reacted components and the unreacted components are separated, and the detection sensitivity is greatly improved; the concentration of the component to be measured in the whole blood sample is accurately converted into the plasma concentration by measuring and calculating the hematocrit or the volume ratio of each sample.
In order to achieve the above object, the present invention provides an automatic detection device for integrated reagents for microfluidic immunoassay, which is characterized in that: the device comprises a working turntable, an automatic sample introduction device, a luminous substrate liquid filling device, a hematocrit scale reader and a fluorescence/luminescence detection unit, wherein the working turntable is divided into an even number of reagent positions, each reagent position can be fixedly provided with a microfluid immunoassay integrated reagent device, a plurality of hematocrit test tube jigs are symmetrically arranged on the working turntable, the hematocrit test tube jigs are arranged to enable the hematocrit test tubes arranged in the hematocrit test tube jigs to change from a vertical position to a horizontal position when the working turntable rotates so as to complete whole blood cell centrifugal precipitation, the automatic sample introduction device is arranged beside a sample introduction station and used for adding a blood sample to be detected into the microfluid immunoassay integrated reagent device transferred to the sample introduction station, the blood sample to be detected is reacted and washed in the microfluid immunoassay integrated reagent device, the luminous substrate liquid filling device is arranged beside a luminous substrate liquid filling station and used for filling luminous substrate liquid into a feature marker reaction tube transferred to the luminous substrate liquid filling station, the hematocrit scale reader is arranged outside the working turntable and used for reading the hematocrit test tube at a fixed position, and the fluorescence/luminescence detection unit is arranged in the luminescence detection unit for detecting the hematocrit test sample and detecting the fluorescence/luminescence detection unit.
Further, the integrated reagent device for microfluid immunoassay comprises a base, an upper cover and a characteristic marker detection reaction tube, wherein a washing liquid storage tank, a marker membrane sample loading liquid guide structure, a reaction tube mounting seat and a waste liquid storage tank are sequentially arranged on the base along the reagent flowing direction, the upper cover is combined with the base in a jogged manner, the characteristic marker detection reaction tube is formed by injection molding of PS (polystyrene) plastics, is fixedly embedded in the reaction tube mounting seat and is provided with a slit-shaped inner cavity so as to generate capillary action to propel the reaction process, a liquid inlet and a liquid outlet are respectively arranged at two ends of the inner cavity, the marker membrane sample loading liquid guide structure fixedly installs a marker membrane in the liquid inlet and is provided with a liquid guide cavity, a washing liquid inlet and a sample inlet which are communicated with the liquid guide cavity, the washing liquid storage tank is communicated with the liquid guide cavity through the washing liquid inlet, the liquid guide cavity is communicated with the liquid inlet through the liquid guide cavity, the waste liquid storage tank is communicated with the liquid outlet through the liquid inlet, and the waste liquid storage tank is communicated with the liquid outlet through the liquid suction material filled in the waste storage tank.
Furthermore, the washing liquid storage tank is an oval cavity structure, a sealed washing liquid bag is placed in the washing liquid storage tank, and an opening is formed in one side of the marker membrane sample loading and leading structure, the opening is in fluid communication with the washing liquid introducing port, a puncture needle is arranged in the opening, and the puncture needle is used for puncturing the washing liquid bag.
Still further, the volume of the wash solution in the wash solution bag is 1 ml.
Furthermore, the marker membrane loading and draining structure comprises a marker membrane fixing clip, and the marker membrane fixing clip is used for pushing and fixing the marker membrane at the marker membrane position of the liquid inlet.
Furthermore, the sample introducing port is of a cross-shaped capping opening and closing structure, the sample is introduced after the sampling needle is ejected, and the opening is closed after the sampling needle is ejected.
Further, the waste reservoir is in fluid communication with the liquid outlet via an isthmus.
Furthermore, a luminescent substrate liquid leading-in hole is formed in the characteristic marker reaction tube close to the liquid outlet, and the luminescent substrate liquid leading-in hole is used for adding luminescent substrate liquid.
Still further, the luminescent substrate liquid filling apparatus includes a luminescent substrate liquid bottle, a luminescent substrate liquid guide tube, a luminescent substrate liquid filling pump, a luminescent substrate liquid filling needle moving motor, and a luminescent substrate liquid filling needle, the luminescent substrate liquid guide tube connects the luminescent substrate liquid bottle and the luminescent substrate liquid filling needle, the luminescent substrate liquid filling pump is mounted on the luminescent substrate liquid guide tube, and the luminescent substrate liquid filling needle moving motor is used to move the luminescent substrate liquid filling needle.
Furthermore, a clamping edge is arranged on the reaction tube mounting seat and used for fixing the characteristic marker reaction tube.
Further, the cross-section of the inner cavity of the characteristic marker reaction tube is circular, rectangular, semicircular or elliptical.
Furthermore, the wall surface of the inner cavity can be provided with an arc-shaped or corrugated structure so as to increase the wall surface area of the inner cavity.
Further, the detection unit includes a detector for detecting fluorescence/luminescence of the reactant in the characteristic marker detection reaction tube, and an excitation light source for projecting light emitted from the excitation light source into the characteristic marker detection reaction tube to excite the reactant to emit fluorescence.
Still further, the detector is a PMT detector.
Furthermore, autoinjection device includes sample cell fixed ring, the rotatory mixing drive structure of sample cell, sample cell reads and records ware, syringe support frame and syringe drive arrangement, the sample cell fixed ring is installed the rotatory mixing drive of sample cell is structural and by its drive rotation in order to fix blood misce bene in the sample cell on the sample cell fixed ring, sample barcode reads and records ware is used for reading bar code on the sample cell is uploaded, the syringe is installed on the syringe support, syringe drive arrangement is used for the drive the syringe support, so as to will the syringe drive is to sample position, application of sample position and washing position, and then makes the syringe implement sample, application of sample and from washing position
Furthermore, the hematocrit test tube jig is provided with a C-shaped cross section, a clamping groove is arranged on the upper surface of the hematocrit test tube jig, a pair of cylindrical protrusions which are radially symmetrical are arranged at the tube opening of the hematocrit test tube, and the cylindrical protrusions are rotationally clamped in the clamping groove.
By adopting the technical scheme, the invention has the beneficial effects that:
(1) PS plastic with high signal-to-noise ratio is used as a solid phase carrier, the reaction process is controlled by utilizing the capillary action and the controllable rotating speed, the reaction is full and complete, the detection is accurate, and the quantification is reliable.
(2) The washing of the reaction tube can be automatically completed, and the sensitivity is obviously improved.
(3) The hematocrit can be detected simultaneously, and the corresponding plasma concentration result can be reported more accurately.
(4) The device has the advantages of simple structure, high automation degree, multi-index detection, arbitrary human number combination detection and high flux.
Drawings
To further illustrate the various embodiments, the invention provides the accompanying drawings. The accompanying drawings, which are incorporated in and constitute a part of this disclosure, illustrate embodiments of the invention and, together with the description, serve to explain the principles of the embodiments. Those skilled in the art will appreciate still other possible embodiments and advantages of the present invention with reference to these figures. The components in the drawings are not necessarily to scale, and similar reference numerals are generally used to identify similar components.
FIG. 1 is a schematic structural diagram of an automatic detection device for integrated reagents of microfluidic immunoassay according to the present invention;
FIG. 2 is a schematic diagram of the structure of the microfluidic immunoassay integrated reagent device of FIG. 1;
FIG. 3 is a schematic view of the structure of the characteristic marker detecting reaction tube in FIG. 2;
FIGS. 4a, 4b, 4c and 4d show different cross-sections of the characteristic marker detection reaction tube described in FIG. 3, respectively.
Detailed Description
The invention will now be further described with reference to the accompanying drawings and detailed description.
As shown in FIG. 1, an automatic detection device for integrated reagents of microfluidic immunoassay may include a working turntable 1, an automatic sample introduction device 2, a luminescent substrate solution filling device 3, a fluorescence/luminescence detection unit 4, and a hematocrit scale reader 7. The working turntable 1, the automatic sample introduction device 2, the luminescent substrate liquid filling device 3, and the fluorescence/luminescence detection unit 4 will be described in detail below.
The working turntable 1 may comprise a rotary shaft 11 driven by a programmable drive motor and a spoke plate 12 fixedly mounted on said rotary shaft 11. The spoke plate 12 is made of engineering plastics through injection molding and integral molding, and is provided with an even number of reagent positions 121 in a partition mode at equal intervals. A sample introduction station A, a luminescent substrate liquid filling station B and a detection station C are arranged in corresponding areas outside the spoke disc 12. The reagent site 121 is provided with a structure which is in mosaic fit with the microfluidic immunoassay integrated reagent device 5 and is tightly fitted and fixed with the microfluidic immunoassay integrated reagent device 5 when the microfluidic immunoassay integrated reagent device 5 is loaded. That is, one microfluidic immunoassay integrated reagent device 5 may be fixedly disposed at each reagent site. For clarity, only two microfluidic immunoassay integrated reagent devices 5 are shown in fig. 1. After the detection items on the loaded microfluidic immunoassay integrated reagent device 5 are detected, the loaded microfluidic immunoassay integrated reagent device 5 can be unloaded, and a new loaded microfluidic immunoassay integrated reagent device 5 to be detected is installed for the next detection operation. The structure of the microfluidic immunoassay integrated reagent-loaded device 5 will be described in detail below. The hematocrit test tube sites 122 are symmetrically distributed on the spider 12, and each hematocrit test tube site 122 is provided with a hematocrit test tube jig (not shown) for loading the hematocrit test tube 6, which is arranged so that the hematocrit test tube 6 placed therein is changed from a vertical position to a horizontal position upon rotation of the working turntable to complete the whole blood cell centrifugal sedimentation. Specifically, the hematocrit test tube jig has a C-shaped cross section, a clamping groove is arranged on the upper surface of the hematocrit test tube jig, a pair of cylindrical protrusions 61 which are radially symmetrical are arranged at the tube opening of the hematocrit test tube 6, and the cylindrical protrusions 61 are rotatably clamped in the clamping groove. That is, the cylindrical protrusion 61 serves as a rotation axis so that the hematocrit test tube 6 can be rotated from a vertical position to a horizontal position by a centrifugal force and the whole blood cell centrifugal sedimentation is completed by the centrifugal force.
A hematocrit scale reader 7 is disposed at a fixed position outside the working turntable for reading the number of hematocytes packed in the hematocrit test tube 6 after centrifugation. The specific construction of the hematocrit scale reader 7 is well known to those skilled in the art and will not be described herein.
The automatic sample introduction device 2 is arranged beside a sample introduction station A and can comprise a sample tube fixing ring 21, a sample tube rotating and uniformly mixing driving structure 22, a sample bar code reader 23, a sample introduction needle 24, a sample introduction needle support frame 25 and a sample introduction needle driving device 26. The syringe fixing ring 21 is mounted on the syringe rotary blending driving structure 22 and can be driven to rotate by the syringe rotary blending driving structure. The sample tube fixing ring 22 firmly fixes the sample tube 27 on the rotation shaft of the sample tube rotational mixing driving structure 22, so as to ensure that the sample tube does not fall off during rotational mixing and sampling. The sample tube rotational mixing drive mechanism 22 is used to mix the whole blood sample prior to sampling to ensure the representativeness of the sample. The sample tube 27 holds samples such as whole blood, serum or plasma, and barcodes attached to the samples can form a one-to-one correspondence with sample information for identification. This barcode should be placed towards the sample barcode reader 23 when the sample tube 27 is loaded so that the sample barcode reader 23 can scan the barcode on the sample tube 27. The sample bar code reader 23 is connected with the upper computer. Because the bar code is linked with the sample information and the index to be detected, the upper computer controls the sampling times and sampling quantity of the sampling needle 24 corresponding to the index to be detected through the bar code information uploaded by the sample bar code reader 23. The injection needle 24 is installed on the injection needle support 25, and the injection needle driving device 26 is used for driving the injection needle support 25 to drive the injection needle 24 to a sampling position, an injection position and a washing position, and further make the injection needle implement sampling, injection and self-washing.
As shown in fig. 2 to 4, the microfluidic immunoassay integrated reagent device 5 may include a base 51, an upper cover (not shown), and a characteristic marker detection reaction tube 52. The base 51 is integrally injection molded by PP plastic, and may include a washing solution reservoir 511, a label film sample loading and liquid guiding structure 512, a reaction tube mounting base 513 and a waste solution reservoir 514, which are sequentially arranged along the reagent flowing direction. The upper cover conforms to the shape of the base 51 and is in embedded engagement with the base 51 to form a closed washing liquid reservoir 511, a label membrane loading drainage structure 512 and a waste liquid reservoir 514. The base 51 is narrower at one end of the lotion reservoir 511 for clarity of orientation. Further, for the convenience of grasping, the middle portion of the base 51 is provided with a recessed portion 515.
As shown in FIG. 2, the washing liquid storage tank 511 has an oval cavity structure, and is provided at one end of the base 51. An opening 5111 is formed at a side of the washing solution storage tank facing the label film loading liquid guide structure 512, and a sharp-edged protrusion is formed in the opening, and a puncture needle 5112 is arranged in the opening. A sealed lotion bag 5113 is placed in the lotion storage tank 511. During centrifugation, the washing solution bag 5113 is pushed to the puncture needle 5112 by centrifugal force, the puncture needle 5112 punctures the washing solution bag 5113, and the washing solution flows out of the washing solution bag 5113 and enters the feature marker detection reaction tube 52 through the opening 5111 and the marker film sample loading liquid guide structure 512, thereby completing the washing function. The washing solution bag 113 contains a washing solution sufficient to wash the characteristic marker detection reaction tube 2 sufficiently. In one embodiment, the volume of the wash solution in the wash solution bag 5113 is about 1 mL.
With continued reference to FIG. 2, a marker film loading wicking structure 512 is located between the wash reservoir 511 and the reaction tube mount 513, and may include a marker film fixing catch 5121, a wicking chamber 5122, a sample introduction port 5123, and a wash introduction port 5124. The marker membrane fixing clip 5121 is used to push the marker membrane 5125 against the marker membrane position in the liquid inlet 5211 of the inner cavity 521 of the characteristic marker detection reaction tube 52, and is further fixed to the marker membrane position by physical structural force. The drainage chamber 5122 is used to allow the wash solution and the sample to be introduced and flow through the marker membrane to the characteristic marker detection reaction tube 52. That is, the drainage chamber 5122 is in fluid communication with the sample introduction port 5123 and the wash solution introduction port 5124. The sample introducing port 5123 is a cross-shaped capping opening and closing structure, the sample is introduced after the sampling needle is ejected, and the opening is closed after the sampling needle is ejected, so that the discharging of the washing liquid during centrifugation is avoided. The washing liquid introduction port 5124 is in fluid communication with a washing liquid storage tank (specifically, the opening 5111), and the washing liquid is introduced into the liquid guide chamber 5122 by high-speed centrifugal force.
As shown in fig. 2 to 4d, the characteristic marker reaction tube 52 is injection molded from PS plastic and is light permeable. Feature marker reaction tube 52 is inserted into reaction tube mount 513. Specifically, the reaction tube mounting base 513 is provided with a clamping edge 5131, and the characteristic marker reaction tube 52 is fixed by the clamping edge 5131. Therefore, the characteristic marker reaction tube 52 does not move upon centrifugation. The characteristic marker reaction tube 52 has a slit-shaped lumen 521, which is a structure that facilitates the generation of capillary action to advance the reaction progress. Namely, the capillary action can be formed in the cavity, which is beneficial to the solid phase process and the driving detection reaction. The cross-section of the lumen 521 may be circular, semi-circular (as shown in fig. 4 a), rectangular (as shown in fig. 4 b), oval (as shown in fig. 4 c), or trapezoidal (as shown in fig. 4 d), etc. The wall 5213 of the inner chamber 521 is a reaction surface, and the upper surface 522 of the feature marker reaction tube 52 is an observation surface. Preferably, the wall surface of the inner cavity 521 is provided with an arc or a corrugated structure to increase the surface area, i.e. the reaction surface area, and further increase the reaction speed. The characteristic marker reaction tube 52 is open at both ends, i.e., the inlet port 5211 and the outlet port 5212 are provided at both ends of the inner cavity 21. The inlet port 5211 is in fluid communication with the drainage lumen 5122 of the label film loading drainage structure 512 and the outlet port 5212 is in fluid communication with the waste reservoir 514. The loading port 5211 is flared for better loading of the marker membrane 5125.
In the illustrated embodiment, the characteristic marker reaction tube 52 is provided with a luminescent substrate liquid introduction hole 523 near the liquid outlet, and the luminescent substrate liquid introduction hole 523 is used for adding luminescent substrate liquid. The luminescent substrate liquid introduction hole 523 is provided with a sealing means (e.g., a sealing film) which is opened when the luminescent substrate liquid is required to be introduced.
The signature reaction tube 52 may also include a protective rim 524, the protective rim 524 being configured to protect the signature reaction tube 52 on the one hand and to cooperate with a retaining rim of the base on the other hand to better secure the signature reaction tube 52.
As shown in fig. 2, the waste reservoir 514 is a hollow structure having a flat horseshoe shape and a closed end. Of course, waste reservoir 514 may take other configurations. The waste liquid reservoir 514 is located at the rear end of the characteristic marker detection reaction tube 52, and is filled with a water absorbing material 5141. The water absorbing material 5141 is used to absorb reaction waste and washing liquid to prevent backflow. The water absorbing material 5141 may be a sponge or a resin, etc. Preferably, waste reservoir 514 is connected to signature detection reaction tube 52 (specifically, outlet 5212) through an isthmus 5142 to prevent backflow of unabsorbed reaction waste and wash solution.
Referring back to FIG. 1, the luminescent substrate liquid filling apparatus 3 includes a luminescent substrate liquid bottle 31, a luminescent substrate liquid guide tube 32, a luminescent substrate liquid filling pump 33, a luminescent substrate liquid filling needle moving motor 34, and a luminescent substrate liquid filling needle 35. The luminescent substrate liquid guide tube 32 is connected with the luminescent substrate liquid bottle 31 and the luminescent substrate liquid filling needle 35, the luminescent substrate liquid filling pump 33 is installed on the luminescent substrate liquid guide tube 32, and the luminescent substrate liquid filling needle moving motor 34 is used for moving the luminescent substrate liquid filling needle 35. When the instruction of adding the luminescent substrate liquid is transmitted to the luminescent substrate liquid adding device 3, the luminescent substrate liquid adding needle moving motor 34 aligns the luminescent substrate liquid adding needle 35 with the luminescent substrate liquid introducing hole 523 of the characteristic marker reaction tube 52 to be added, the luminescent substrate liquid adding pump 33 is started, and the luminescent substrate liquid adding needle 35 injects the luminescent substrate liquid of a set volume into the reaction tube. The working turntable 1 rotates one station and the same luminescent substrate liquid is injected into the other feature marker reaction tube 52 in the same manner. And so on until all the characteristic marker reaction tubes 52 finish the sample adding of the luminescent substrate solution.
The detection unit 4 includes a detector 41 and a fluorescence excitation light source 42. The detector 41 is used to detect the fluorescence/luminescence of the reactants in the characteristic marker detection reaction tube 52. The fluorescence excitation light source 42 is used to excite fluorescence of the reactants in the feature marker detection reaction tubes 52. In the case where the luminescent substrate liquid is added to the feature marker detection reaction tube 52, the fluorescence excitation light source 42 is not required to be used. The fluorescence/luminescence intensity is related to the concentration of the target to be detected. And establishing a standard curve according to the fluorescence/luminescence signal intensity and the target object concentration, and then realizing qualitative and quantitative detection of the sample target object. If the fluorescence/luminescence signal intensity is detected in sections in the reaction area, the accumulated fluorescence/luminescence intensity represents the amount of the target substance combined in the reaction area; and (3) calculating in a segmented mode, and conjecturing the amount of each segment, such as the extremely weak fluorescence/luminescence of the last segment, which represents the complete reaction, such as the strong fluorescence/luminescence of the last segment, which represents the incomplete reaction, and many target material positions still participate in the reaction, and the dilution and quantification are needed.
The working principle of the present invention is explained in detail below.
First, the information of each sample and the name of each sample detection index required for the test are set, the microfluidic immunoassay integrated reagent device 5 corresponding to each sample detection index is positioned on the reagent site 121 of the work turntable 1, and the hematocrit test tube 6 containing a predetermined volume of whole blood is loaded on the hematocrit test tube site 122 of the work turntable 1. The microfluidic immunoassay integrated reagent device 5 of a certain sample is rotated to a sample injection position.
Next, the sample to be detected is placed in the sample tube 27 and placed on the sample tube fixing ring 21, and the barcode on the sample tube 27 corresponds to the detection surface of the sample barcode reader 23. After the sampling detection information is sent out, the sample tube rotating and mixing driving structure 22 rotates and mixes the sample tube uniformly for one to several times, the sample tube is static and stands up, the sample bar code reader 23 scans the bar code information on the sample tube, the automatic sample feeding device 2 sequentially feeds the microfluid immunoassay integrated reagent device corresponding to the item to be detected according to the bar code information, until the whole sample feeding of the item to be detected of one sample is finished, then the sample feeding needle 23 is washed, the microfluid immunoassay integrated reagent device corresponding to the detection index required by the 2 nd sample is started to sequentially feed the sample, and the like.
Then, the sample applied to the area of the marker film 5125 dissolves the marker film needle 5112, the needle 5112 pierces the marker on the washing solution bag 5113, and the complex formed by the target and the marker in the sample is transferred to the inner cavity of the reaction tube 52 by means of capillary force, and reacts with the solid phase on the reaction surface 5213 to form a reaction detection chain. The reaction is continued for a certain time by the action of capillary force or centrifugal force until the reaction surface 5213 of the reaction tube 52 is completely covered with the reaction liquid and continued for a predetermined time.
An intermittent low-speed centrifugation is arranged to accelerate the reaction process so that the reactants fully reach the whole reaction surface 5213. Part of the reactant flows into the waste liquid storage tank 514 of the microfluidic immunoassay integrated reagent device 5, and is absorbed and retained by the water absorbing material 5141 in the waste liquid storage tank 514.
After the specified reaction time is complete, the washing process is started. The disc 12 is driven to rotate at a preset rotation speed for a predetermined time period in accordance with information such as time and rotation speed set by the control program. At this time, the washing solution bag 5113 in the integrated microfluidic immunoassay reagent device 5 is pushed against the puncture needle 5112 in the integrated microfluidic immunoassay reagent device 5 by the centrifugal force, the puncture needle 5112 punctures the washing solution bag 5113, and the washing solution flows along the predetermined liquid path to the marker film 5125 and the reaction tube 52 of the integrated microfluidic immunoassay reagent device, finally flows to the waste liquid tank 514, is absorbed and retained by the water absorbent material 5141 in the waste liquid tank 514, and finally washes the marker film 5125 and the reaction tube 52 clean.
The reacted and washed microfluid immunoassay integrated reagent device 5 sequentially rotates to a luminescent substrate liquid filling station B according to a preset sequence. When the instruction of adding the luminescent substrate liquid is transmitted to the luminescent substrate liquid adding device 3, the luminescent substrate liquid adding needle moving motor 34 aligns the luminescent substrate liquid adding needle 35 with the luminescent substrate liquid introducing hole 523 of the characteristic marker reaction tube 52 to be added with the sample, the luminescent substrate liquid adding pump 33 is started, and the luminescent substrate liquid adding needle 35 injects the luminescent substrate liquid of a set volume into the reaction tube. The working turntable 1 rotates one station and the same luminescent substrate liquid is injected into the other feature marker reaction tube 52 in the same manner. And the same can be done until all the feature marker reaction tubes 52 finish the loading of the luminescent substrate solution. It should be noted that, in the case of fluorescence detection, addition of a luminescent substrate solution is not required, and therefore, the addition of the luminescent substrate solution is omitted.
And the microfluid immunoassay integrated reagent device after reaction and washing or after adding luminescent substrate liquid sequentially rotates to a detection station C according to a preset sequence. In the case of fluorescence detection, the fluorescence excitation light source 42 of the fluorescence/luminescence detection unit excites the reaction surface 5213 of the feature marker reaction tube 52 to generate fluorescence, the detector scans the reaction surface 5213 in a segmented manner from near to far, measures the fluorescence/luminescence values of each segment, and calculates the fluorescence/luminescence values of each segment and the ratio of the fluorescence/luminescence values of each segment. In the case where a luminescent substrate solution is added (i.e., luminescence detection), the light emission value is directly detected by the pmt without exciting the fluorescence excitation light source 42.
After the fluorescence/luminescence test is finished, the working turntable 1 rotates at high speed for 3 minutes again, the whole blood in the hematocrit test tube 6 continues to be centrifugally precipitated, the hematocrit scale reader 7 measures the occupied height of the plasma and the blood cells in the hematocrit test tube after the whole blood is static, and the hematocrit of each sample is calculated for calculating the concentration of the target substances in the plasma and the whole blood. According to the established and stored standard curve, calculating the concentration value of the whole blood sample corresponding to each representative index of the microfluidic immunoassay integrated reagent device, and then converting the plasma concentration value according to the hematocrit.
While the invention has been particularly shown and described with reference to a preferred embodiment, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (8)
1. An integrated reagent automatic checkout device of microfluid immunoassay which characterized in that: comprises a working turntable, an automatic sample introduction device, a luminescent substrate liquid filling device, a hematocrit scale reader and a fluorescence or luminescence detection unit, the working turnplate is equally divided into an even number of reagent positions, each reagent position can be fixedly provided with a microfluid immunoassay integrated reagent device, the working turntable is symmetrically provided with a plurality of hematocrit test tube jigs which are arranged to change the hematocrit test tubes placed in the hematocrit test tube jigs from vertical positions to horizontal positions when the working turntable rotates so as to complete the whole blood cell centrifugal precipitation, the hematocrit test tube jigs are provided with C-shaped cross sections, the upper surface of the test tube is provided with a clamping groove, the opening of the hematocrit test tube is provided with a pair of cylindrical bulges which are symmetrical in the radial direction, the cylindrical protrusion is rotationally clamped in the clamping groove, the automatic sample feeding device is arranged beside the sample feeding station, is used for adding a blood sample to be detected into the microfluidic immunoassay integrated reagent device transferred to the sample feeding station, the blood sample to be detected is reacted and washed in the microfluidic immunoassay integrated reagent device, the luminescent substrate liquid filling device is arranged beside the luminescent substrate liquid filling station, used for filling the luminescent substrate liquid into the characteristic marker reaction tube transferred to the luminescent substrate liquid filling station, the hematocrit scale reader is arranged at a fixed position outside the working turntable, for reading the number of blood cell volumes in the centrifuged hematocrit test tube, the fluorescence or luminescence detection unit being arranged at a detection station, the device is used for carrying out fluorescence or luminescence detection on the reacted and washed sample in the microfluidic immunoassay integrated reagent device transferred to the detection station; the microfluid immunoassay integrated reagent device comprises a base, an upper cover and a characteristic marker detection reaction tube, wherein a washing liquid storage tank, a marker film sample loading liquid guide structure, a reaction tube mounting seat and a waste liquid storage tank are sequentially arranged on the base along the reagent flowing direction, the upper cover is combined with the base in a tabling mode, the characteristic marker detection reaction tube is formed by PS plastic in an injection molding mode, is fixedly embedded in the reaction tube mounting seat, is provided with an observation surface facing upwards and is provided with a slit-shaped inner cavity so as to generate capillary action to propel the reaction process, two ends of the inner cavity are respectively provided with a liquid inlet and a liquid outlet, the marker film sample loading liquid guide structure fixedly installs the marker film in the liquid inlet and is provided with a liquid guide cavity, a washing liquid inlet and a sample inlet which are communicated with the liquid guide cavity, the washing liquid storage tank is communicated with the liquid guide cavity through the washing liquid inlet, the liquid guide cavity is communicated with the liquid inlet, the waste liquid storage tank is communicated with the liquid outlet through fluid, and water absorption materials are filled in the waste liquid storage tank.
2. The microfluidic immunoassay integrated reagent automatic detection device of claim 1, wherein: the washing liquid storage tank is of an oval cavity structure, a sealed washing liquid bag is placed in the washing liquid storage tank, an opening is formed in one side, facing the sample loading and liquid guiding structure of the marker membrane, of the washing liquid storage tank, the opening is in fluid communication with the washing liquid introducing port and is internally provided with a pricking pin, and the pricking pin is used for pricking the washing liquid bag.
3. The microfluidic immunoassay integrated reagent automatic detection device of claim 1, wherein: the sample introducing port is a cross-shaped capping opening and closing structure, a sample is introduced after the sampling needle is ejected, and the opening is closed after the sampling needle is ejected.
4. The microfluidic immunoassay integrated reagent automatic detection device according to claim 1, wherein: the waste reservoir is in fluid communication with the liquid outlet via a isthmus.
5. The microfluidic immunoassay integrated reagent automatic detection device of claim 1, wherein: and a luminescent substrate liquid leading-in hole is formed in the characteristic marker reaction tube close to the liquid outlet, and is used for adding luminescent substrate liquid.
6. The microfluidic immunoassay integrated reagent automatic detection device of claim 5, wherein: the luminous substrate liquid filling device comprises a luminous substrate liquid bottle, a luminous substrate liquid guide pipe, a luminous substrate liquid filling pump, a luminous substrate liquid filling needle moving motor and a luminous substrate liquid filling needle, wherein the luminous substrate liquid guide pipe is connected with the luminous substrate liquid bottle and the luminous substrate liquid filling needle, the luminous substrate liquid filling pump is installed on the luminous substrate liquid guide pipe, and the luminous substrate liquid filling needle moving motor is used for moving the luminous substrate liquid filling needle.
7. The microfluidic immunoassay integrated reagent automatic detection device according to claim 1, wherein: the automatic sampling device comprises a sample tube fixing ring, a sample tube rotating mixing driving structure, a sample bar code reading and recording device, a sampling needle support frame and a sampling needle driving device, wherein the sample tube fixing ring is installed on the sample tube rotating mixing driving structure and is driven to rotate by the sample tube rotating mixing driving structure so as to be fixed, a blood sample in a sample tube on the sample tube fixing ring is uniformly mixed, the sample bar code reading and recording device is used for reading a bar code on the sample tube and uploading, the sampling needle is installed on the sampling needle support, the sampling needle driving device is used for driving the sampling needle support so as to drive the sampling needle to a sampling position, a sampling position and a washing position, and further make the sampling needle implement sampling, sampling and self-washing.
8. The microfluidic immunoassay integrated reagent automatic detection device according to claim 1, wherein: the detection unit comprises a detector and an excitation light source, light emitted by the excitation light source is projected into the characteristic marker detection reaction tube to excite the reactant to emit fluorescence, and the detector is used for detecting the fluorescence or luminescence of the reactant in the characteristic marker detection reaction tube.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910217642.XA CN109975564B (en) | 2019-03-21 | 2019-03-21 | Automatic detection device for microfluid immunoassay integrated reagent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910217642.XA CN109975564B (en) | 2019-03-21 | 2019-03-21 | Automatic detection device for microfluid immunoassay integrated reagent |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109975564A CN109975564A (en) | 2019-07-05 |
| CN109975564B true CN109975564B (en) | 2022-12-13 |
Family
ID=67079986
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910217642.XA Active CN109975564B (en) | 2019-03-21 | 2019-03-21 | Automatic detection device for microfluid immunoassay integrated reagent |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109975564B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110967307A (en) * | 2019-11-22 | 2020-04-07 | 宁波紫园医疗器械有限公司 | Hematocrit measuring device |
| CN113419073B (en) * | 2020-03-30 | 2024-05-31 | 厦门先明生物技术有限公司 | High-flux full-automatic microfluidic fluorescence immunoassay device |
| CN113281499A (en) * | 2021-05-18 | 2021-08-20 | 厦门先明生物技术有限公司 | Microfluidic immunoassay joint detection device and using method thereof |
| CN117871853B (en) * | 2024-03-13 | 2024-05-24 | 南通戴尔诺斯生物科技有限公司 | Colloidal gold immunochromatography equipment for detecting biotoxin and application method thereof |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102680441A (en) * | 2011-03-16 | 2012-09-19 | 罗姆股份有限公司 | Analysis chip, analysis system, and analysis method |
| US10493448B2 (en) * | 2015-03-13 | 2019-12-03 | Bio-Rad Laboratories, Inc. | Assay cartridge |
| CN104698199A (en) * | 2015-04-01 | 2015-06-10 | 山东美医林电子仪器有限公司 | Hemorheology detecting device |
| CN107102131B (en) * | 2017-07-03 | 2019-02-05 | 沈阳微流控生物科技有限公司 | A kind of full-automatic micro-fluidic chip fluorescence immunoassay detection system and its detection method |
| CN109061207A (en) * | 2018-06-21 | 2018-12-21 | 微粒云科技（北京）有限公司 | A kind of biochip for heart infarction marker detection, detection method |
-
2019
- 2019-03-21 CN CN201910217642.XA patent/CN109975564B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN109975564A (en) | 2019-07-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109975564B (en) | Automatic detection device for microfluid immunoassay integrated reagent | |
| EP3572815B1 (en) | Automatic analysis device and sample analysis method | |
| CN1124489C (en) | Analysis system | |
| US6299839B1 (en) | System and methods for performing rotor assays | |
| US9164091B2 (en) | Centrifugal micro-fluidic device and method for detecting analytes from liquid specimen | |
| JP2539512B2 (en) | Multi-item analyzer and method for operating the analyzer | |
| US9442109B2 (en) | Centrifugal force based microfluidic system and method for the automated analysis of samples | |
| Schembri et al. | Centrifugation and capillarity integrated into a multiple analyte whole blood analyser | |
| CA2392943C (en) | Chemistry system for a clinical analyzer | |
| US7700042B2 (en) | Automated analyzer | |
| JP4339879B2 (en) | Sample collection and weighing device with integrated liquid compartment | |
| JP2842758B2 (en) | Automatic analyzer | |
| JP2006518449A (en) | Microfluidic biochip with a breakable seal | |
| CN109975563B (en) | Integrated reagent device for microfluid immunoassay | |
| WO1990010875A1 (en) | Analyzer of liquid sample and analyzing method of liquid sample using said analyzer | |
| MXPA06000249A (en) | Automated multi-detector analyzer. | |
| CA1298181C (en) | Unitized reagent containment system for clinical analyzer | |
| CN107843583A (en) | A kind of micro-fluidic chemiluminescent analyzer and application method | |
| CN110354924A (en) | A kind of integrated bio pattern detection chip and detection method | |
| CN207380068U (en) | A kind of micro-fluidic chemiluminescent analyzer | |
| JPH0623767B2 (en) | Method and apparatus for analyzing liquid sample | |
| JP2004309233A (en) | Chemical analysis system | |
| US20060078471A1 (en) | Apparatus and method for a precision flow assay | |
| US20060079003A1 (en) | Apparatus and method for a precision flow assay | |
| CN114453037A (en) | Homogeneous test micro-fluidic chip and detection system |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |