WO2015127656A1 - Dispositif d'essai du virus d'immunodéficience humaine et son procédé d'essai - Google Patents

Dispositif d'essai du virus d'immunodéficience humaine et son procédé d'essai Download PDF

Info

Publication number
WO2015127656A1
WO2015127656A1 PCT/CN2014/072725 CN2014072725W WO2015127656A1 WO 2015127656 A1 WO2015127656 A1 WO 2015127656A1 CN 2014072725 W CN2014072725 W CN 2014072725W WO 2015127656 A1 WO2015127656 A1 WO 2015127656A1
Authority
WO
WIPO (PCT)
Prior art keywords
sample
pool
detected
detecting
immunodeficiency virus
Prior art date
Application number
PCT/CN2014/072725
Other languages
English (en)
Chinese (zh)
Inventor
郭文鹏
蔡志明
Original Assignee
深圳市第二人民医院
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by 深圳市第二人民医院 filed Critical 深圳市第二人民医院
Priority to PCT/CN2014/072725 priority Critical patent/WO2015127656A1/fr
Publication of WO2015127656A1 publication Critical patent/WO2015127656A1/fr

Links

Images

Classifications

    • 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
    • G01N35/0098—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502761—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L7/00—Heating or cooling apparatus; Heat insulating devices
    • B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/70—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving virus or bacteriophage
    • C12Q1/701—Specific hybridization probes
    • C12Q1/702—Specific hybridization probes for retroviruses
    • C12Q1/703—Viruses associated with AIDS
    • 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
    • G01N35/00069—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 whereby the sample substrate is of the bio-disk type, i.e. having the format of an optical disk
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0647—Handling flowable solids, e.g. microscopic beads, cells, particles
    • B01L2200/0668—Trapping microscopic beads
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0816—Cards, e.g. flat sample carriers usually with flow in two horizontal directions
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
    • B01L2300/087—Multiple sequential chambers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/18—Means for temperature control
    • B01L2300/1838—Means for temperature control using fluid heat transfer medium
    • B01L2300/1844—Means for temperature control using fluid heat transfer medium using fans
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/043—Moving fluids with specific forces or mechanical means specific forces magnetic forces
    • 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
    • G01N2035/00099—Characterised by type of test elements
    • G01N2035/00158—Elements containing microarrays, i.e. "biochip"
    • 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
    • G01N2035/00346—Heating or cooling arrangements
    • G01N2035/00356—Holding samples at elevated temperature (incubation)
    • G01N2035/00366—Several different temperatures used

Definitions

  • the invention belongs to the technical field of biochemical analysis, and particularly relates to a human immunodeficiency virus detecting device and a detecting method for detecting human immunodeficiency virus by using the detecting device.
  • AIDS Acquired Immunodeficiency Syndrome Syndrome
  • Human human immunodeficiency virus
  • HIV human immunodeficiency virus
  • the main routes of transmission of HIV include blood transmission, sexual contact, drug injection and mother-to-child transmission.
  • the incubation period in the human body averages 7 to 10 years.
  • Patients with HIV infection have no obvious clinical symptoms, but they are highly contagious.
  • HIV detection to find the source of infection and cut off the route of transmission is the most effective means of preventing and treating AIDS.
  • the most effective and widely used method for HIV immunology detection, HIV antibody screening is the legal four blood-based screening projects in China. One.
  • the most common method for detecting HIV antibodies is to use a detection kit.
  • the sample to be tested is applied from the sample hole to the sample pad of the test strip, so that the sample dissolves the labeled magnetic beads and is chromatographed on the NC membrane, and then The magnetic detector is used to detect the data on the test strip within a specified time, and the result is determined based on the read data.
  • the detection kit has problems such as low detection automation, large reagent consumption, and low detection sensitivity. Meet the requirements for rapid, accurate and efficient detection of early diagnosis of human immunodeficiency virus.
  • the object of the present invention is to provide a human immunodeficiency virus detecting device, which aims to solve the problem that the detecting device provided by the prior art has low detection automation, large reagent consumption, and low detection sensitivity.
  • the invention is realized by a human immunodeficiency virus detecting device for detecting a sample to be detected, comprising a support member and a microfluidic chip disposed on the support member, wherein the microfluidic chip is provided with a sample processing chamber in which the sample to be detected and the magnetic microbead are placed, a sample processing region in which the sample to be detected is sequentially subjected to adsorption, washing, and aggregation treatment, and the sample to be detected processed through the sample processing region a sample signal amplification tank in which the sample is subjected to signal amplification processing, a sample detection pool connected to the sample signal amplification tank, and a sample signal amplification tank and the sample detection pool are disposed between the sample signal amplification tank and the sample signal amplification tank a detecting element for detecting the sample to be tested, the sample pool, the sample processing area and the sample signal amplifying pool are sequentially disposed on the sampling channel and connected; the support member is further provided with a driving device
  • control device includes a circuit board disposed on the support member and a control component disposed on the circuit board, and the circuit board is provided with an installation conforming to the shape of the microfluidic chip a slot, the microfluidic chip is detachably mounted in the mounting slot.
  • the driving device includes a stepping motor disposed under the microfluidic chip, the stepping motor is electrically connected to the control component, and an output shaft of the stepping motor is provided with the magnetic Magnetic material that is magnetically attracted by the microbeads.
  • one side of the circuit board is provided with a heat dissipation fan for assisting the temperature adjustment device to cool down, the heat dissipation fan is electrically connected to the control component, and an air outlet of the heat dissipation fan faces the microfluidic chip. .
  • the support member is further provided with an alternating current converter that provides a DC power supply required by the control element, and the alternating current converter is electrically connected to the control element.
  • the sample processing area includes a sample adsorption zone for performing adsorption processing on the sample to be detected, a sample washing zone for performing washing treatment on the sample to be detected, and a sample aggregation zone for performing aggregation processing on the sample to be detected.
  • the sample adsorption zone, the sample washing zone, and the sample collection zone are sequentially connected through the sampling channel.
  • the microfluidic chip is composed of a substrate and a cover sheet covered on the substrate, the sample bath, the sample adsorption area, the sample washing area, the sample collection area, and the The sample signal amplification cell, the sample detection cell, and the sample channel are respectively etched on the substrate.
  • a buffer solution pool for placing a buffer is further etched on the substrate, and the buffer solution pool and the sample detection pool are divided into two sides of the sample signal amplification tank, and the sample signal amplification pool is Connected.
  • the detecting component is an electrophoresis detector
  • the electrophoresis measuring instrument has a first electrode and a second electrode
  • the first electrode is disposed in the sample signal amplification pool
  • the second electrode is disposed in the In the sample detection pool
  • an electrophoresis detection channel is formed between the sample signal amplification pool and the sample detection pool.
  • the human immunodeficiency virus detecting device provided by the present invention has the beneficial effects that the processing procedure of the sample to be detected is integrated on the microfluidic chip, and the detecting person places the sample to be detected into the sampling pool, and activates the detecting device.
  • the detecting device performs automatic operations such as cell lysis, adsorption, washing, aggregating, signal amplification, and detection on the sample to be tested, thereby reducing the workload of the testing personnel and improving the detection sensitivity.
  • Another object of the present invention is to provide a method for detecting human immunodeficiency virus using a human immunodeficiency virus detecting device as described above, comprising the following detecting steps:
  • the control device starts the operation of the driving device, the driving device drives the magnetic microbeads to move to the sample processing region, and the sample to be detected is adsorbed, washed and aggregated in the sample processing region. reaction;
  • the driving device drives the magnetic microbead into the sample signal amplification cell, and the sample to be detected undergoes a polymerase chain reaction in the sample signal amplification cell to generate a nucleic acid. molecule;
  • the detecting element detects the nucleic acid molecule.
  • the invention has the beneficial effects that the detecting method only needs to place the sample to be tested and the lysing liquid in the sampling pool, and the rest of the operations are automatically completed by the device, the operation is simple, the degree of automation is high, and the workload of the testing personnel is reduced, and The detection sensitivity is high.
  • FIG. 1 is a perspective view of a human immunodeficiency virus detecting apparatus according to an embodiment of the present invention
  • FIG. 2 is a front view of a human immunodeficiency virus detecting apparatus according to an embodiment of the present invention
  • FIG. 3 is a schematic structural diagram of a microfluidic chip for a human immunodeficiency virus detecting apparatus according to an embodiment of the present invention.
  • the present invention provides a human immunodeficiency virus detecting device for detecting a sample to be detected, comprising a support member 1 and a microfluidic chip 2, and the microfluidic chip 2 is disposed on the support member 1
  • the microfluidic chip 2 is provided with a sample introduction chamber 21 for placing a sample to be detected and a magnetic microbead (not shown), and a sample processing area 22 for sequentially performing adsorption, washing, and aggregation processing on the sample to be detected, and
  • the sample signal amplifying pool 23 and the sample detecting pool 24 for performing signal amplification processing on the sample to be detected processed by the sample processing area 22, the sample signal amplifying pool 23 and the sample detecting pool 24 are connected, and the sample signal amplifying pool 23 and the sample detecting pool 24 are connected.
  • a detecting element (not shown) for detecting the sample to be tested processed by the sample signal amplifying pool 23 is provided between the sampling cell 21, the sample processing area 22, and the sample signal amplifying pool 23, which are sequentially disposed in the injection.
  • the channel 25 is connected to the same, and the supporting member 1 is further provided with a driving device 3 for driving the magnetic microbeads to enter the sample signal amplifying pool 23 from the sampling pool 21 along the sampling channel 25, and the support member 1 is provided with temperature regulation. 4
  • the sample to be detected undergoes polymerase chain reaction under the action of the temperature adjusting device 4, and the polymerase chain reaction is to use the DNA to become single-stranded at a high temperature of 95 ° C in vitro, and the temperature is low (usually 60 °).
  • the primers When C is around, the primers are combined with the single strands according to the principle of complementary pairing with bases, and then the temperature is adjusted to the optimum temperature of DNA polymerase (about 72 ° C), and the DNA polymerase is along the phosphoric acid to five carbon sugars (5'-3).
  • the direction of ') synthesizes a complementary chain, and the temperature adjusting device 4 provided by the embodiment of the present invention can perform good control between the denaturation temperature, the refolding temperature, and the extension temperature.
  • the support member 1 is further provided with a control device (not shown) for controlling the driving device 3, the temperature adjusting device 4, and the detecting component, and the detecting person places the sample to be tested on the injection.
  • a control device for controlling the driving device 3, the temperature adjusting device 4, and the detecting component, and the detecting person places the sample to be tested on the injection.
  • a lysate is added into the sample cell 21, and the sample to be tested is subjected to cell lysis, the cell membrane of the sample to be tested is broken, and a small molecular protein fragment is decomposed, and then the coating is added to the sample cell 21 with specificity.
  • the magnetic microbeads of the envelope protein antibody and the sample treatment solution, the protein fragments are anti-binding with the magnetic microbead-coated specific envelope protein antibody, so when the magnetic microbeads move, the small molecular protein fragments are driven for sample processing. .
  • the control device controls the operation of the driving device 3, and the driving device 3 drives the magnetic microbeads adsorbed with the small molecular protein fragments to rotate into the sample processing region 22, and the sample buffer in the sample to be detected and the sample processing region 22
  • the sample is adsorbed, washed, aggregated, etc., and after the above reaction is completed, the driving device 3 is operated again to move the magnetic microbeads into the sample signal amplifying tank 23, and the control device controls the temperature adjusting device 4 to operate, and the temperature adjusting device 4 treats
  • the sample is tested for temperature cycling operation, and the operation includes the following steps: a, heating to 94 ° C for 2 min; b, 94 ° C for 30 sec; c, cooling to 55 ° C for 30 sec; d, warming to 72 ° C for 30 sec; e, 72 ° C for 3 min; f.
  • Step b to step e are cycled 20-25 times, providing the temperature required for the polymerase chain reaction of the sample to be detected, ensuring that the sample to be detected generates enough nucleic acid molecules, and the detecting element detects the nucleic acid molecule to obtain the detection result.
  • the automatic operation is adopted, the operation is simple, the workload of the tester is reduced, the detection sensitivity of the sample to be tested is improved, and the amount of reagent used is small, and the reagent consumption is reduced.
  • the support member 1 provides support for the driving device 3, the temperature adjusting device 4, and the control device, and the support member 1 can be a work table, a support frame, or the like.
  • the composition of the sample lysate is 50 mM Tris (pH 7.4), 150 mM NaI, 1%. NP-40, 0.1% SDS, 0.1% EDTA.
  • the main components of the cell lysate are further detailed below:
  • NP-40 It is a detergent. In biological experiments, it is commonly used in the middle of cell lysates. The cell membrane can be destroyed, but the nuclear membrane is retained. NP-40 is a very mild detergent. The 1% concentration can basically destroy the membrane, but the effect on the destruction of the nuclear membrane is weak. The cytoplasmic protein can be obtained by binding to a specific buffer.
  • SDS sodium dodecyl sulfonate
  • SDS sodium dodecyl Sulfate
  • SDS is the main ingredient of detergent. It is commonly used in DNA extraction to separate proteins from DNA after denaturation. SDS is a known detergent that denatures proteins. It is used to determine the molecular weight of polyacrylamide gel electrophoresis. It can also be used to disrupt cell walls and cleave nucleic acid-protein complexes in nucleic acid extraction operations. At higher temperatures, the binding of proteins to DNA is disrupted and the DNA is released.
  • EDTA It is a good complexing agent in chemistry. It has six coordinating atoms. The complex formed is called chelate. EDTA is often used in coordination titration. Generally, the content of metal ions is determined. In applications, it is used to exclude most of the interference of excessive metal element ions (such as iron (III), nickel (II), manganese (II)). Interfering ions can be removed in protein engineering and testing without affecting protein function.
  • metal element ions such as iron (III), nickel (II), manganese (II)
  • composition of the sample treatment solution is: 0.05M Na 2 CO 3 -NaHCO 3 (PH 9.6 )
  • the control device includes a circuit board 51 disposed on the support member 1 and a control component 52 disposed on the circuit board 51.
  • the circuit board 51 is provided with an outer shape of the microfluidic chip 2.
  • the mounting slot 511, the microfluidic chip 2 is detachably mounted in the mounting slot 511. Since the microfluidic chip 2 is a disposable product, the microfluidic chip 2 is prevented from being used interchangeably. Therefore, each case of the human immunodeficiency virus is completed. In the detection, the microfluidic chip 2 should be taken out from the mounting slot 511 in time, and the new microfluidic chip 2 is replaced in time, and the universal microfluidic chip 2 is used, which is easy to operate.
  • the driving device 3 includes a stepping motor 31 disposed under the microfluidic chip 2.
  • the stepping motor 31 is electrically connected to the control element 52, and the output shaft of the stepping motor 31 is provided with a circle.
  • a disk (not shown), the outer diameter of the disk is adapted to the outer diameter of the injection channel 25, and the disk is provided with a magnetic member corresponding to the magnetic microbead (not shown)
  • the magnetic member is disposed directly under the injection pool 21 and is attracted to the magnetic microbead.
  • the magnetic microbeads move into the sample processing zone 22, and then the sample to be tested is processed.
  • the operation of the stepping motor 31 is controlled by the control element 52, without manual operation by the inspector, high degree of automation, compact space, and magnetic parts and magnetic micro There is no direct contact between the beads, which can avoid contamination of the sample and ensure the accuracy of the test.
  • one side of the circuit board 51 is provided with a cooling fan 6 for assisting the temperature adjustment device 4 to cool down.
  • the cooling fan 6 is electrically connected to the control element 52, and the air outlet of the cooling fan 6 faces the microfluidic chip. 2, since the temperature adjusting device 4 needs to cyclically heat and cool the microfluidic chip 2, for faster assisting and rapid cooling, the cooling fan 6 is turned on, and when the temperature adjusting device 4 is warmed up, the control element 52 automatically turns off the heat dissipation.
  • the fan 6 is convenient and quick to implement without the need for inspection personnel.
  • the support member 1 is further provided with an AC power converter 7 for supplying a DC power supply required for the control element 52.
  • the AC power converter 7 is electrically connected to the control element 52. Since the control element 52 is powered by DC, the AC power is The converter 7 can convert the mains 220v into the DC power required by the control element 52, ensuring that the control element 52 is continuously powered and the human immunodeficiency virus detection device is functioning properly.
  • the sample processing area 22 includes a sample adsorption zone 221 for performing adsorption processing on the sample to be detected, a sample washing zone 222 for performing a washing process on the sample to be detected, and a sample aggregating zone 223 for performing aggregation processing on the sample to be detected,
  • the sample adsorption zone 221, the sample washing zone 222, and the sample aggregating zone 223 are sequentially connected through the injection channel 25, and the injection channel 25 is arranged in a circular shape, and the injection pool 21, the sample adsorption zone 221, the sample washing zone 222, and the sample accumulation zone are provided.
  • the control element 52 controls the operation of the stepping motor 31 again.
  • the magnetic microbeads are moved to the sample washing zone 222, the sample washing liquid is provided in the sample washing zone 222, the sample washing liquid is washed, and the stepping motor 31 drives the magnetic microbeads again into the sample collecting zone 223. After standing for 2 ⁇ 3 minutes, the samples to be tested are gathered, and waiting for the next operation, the operations of adsorption, washing and aggregation of the samples to be tested are all controlled by the control element 52.
  • the system realizes a small amount of reagent loss and a high degree of automation.
  • the sample washing liquid component is: phosphate buffer 0.01 M, pH 7.2
  • the microfluidic chip 2 is composed of a substrate 20 and a cover sheet (not shown) which is disposed on the substrate 20.
  • the sample introduction chamber 21 and the sample adsorption region are formed by CO2 laser lithography. 221, the sample washing area 222, the sample collecting area 223, the sample signal amplifying pool 23, the sample detecting pool 24, and the sampling channel 25 are respectively etched on the substrate 21, in order to make the sample injection tank 21, the sample adsorption area 221, and the sample
  • the washing zone 222, the sample collecting zone 223 and the sample signal amplifying pool 23 are arranged on the substrate 20 and facilitate the transfer of the HIV virus sample between the functional units, the sample injection tank 21, the sample adsorption zone 221, the sample washing zone 222,
  • the sample collection area 223 and the sample amplification tank 23 are arranged in the same circumferential direction so that the sample to be tested is more fluid.
  • a buffer solution pool 26 for arranging a buffer is further etched on the substrate 20.
  • the buffer solution pool 26 and the sample detection tank 24 are separated from both sides of the sample signal amplification tank 23, and the sample signal amplification tank 23 is provided.
  • a buffer solution is placed in the solution buffer tank 26, and the buffer solution dilutes the sample to be tested to facilitate the next detection.
  • the detecting component is an electrophoresis detector
  • the electrophoresis measuring instrument has a first electrode and a second electrode, the first electrode is disposed in the sample signal amplifying pool 23, and the second electrode is disposed in the sample detecting pool 24, and the sample is An electrophoresis detection channel is formed between the signal amplification pool 23 and the sample detection pool 24.
  • the electrophoresis detector is powered on, the electrophoresis enrichment of the sample to be detected can be realized, and a certain excitation signal is applied to monitor the change of the first electrode and the second electrode.
  • the corresponding electrical response signal realizes the quantitative detection of the sample to be detected by the microfluidic chip 2, and establishes a rapid and accurate quantitative detection and analysis of HIV.
  • an ultraviolet light source is arranged above the electrophoresis detection channel, and under ultraviolet irradiation, it can be The associated nucleic acid molecules are identified, and the tester performs diagnosis based on the displayed results to improve the diagnostic accuracy.
  • the HIV inactivated virus P24 envelope protein is used as a test object:
  • the sample to be tested is placed in the injection pool 21, and the lysate is added into the injection pool 21 to cause the sample to be detected to undergo cell lysis, and then the surface coated with the specific envelope protein antibody is added to the injection tank 21. Magnetic microbeads and sample treatment solution, and allowed to stand for 10 to 15 minutes;
  • the control device activates the operation of the driving device 3.
  • the driving device 3 drives the magnetic microbeads to move to the sample processing region 22.
  • the sample to be detected is adsorbed, washed, and aggregated in the sample processing region 22, specifically, the sample to be detected is moved.
  • the body moves to the sample washing zone 222 and is allowed to stand for a few minutes to cause the sample to be detected to interact with the sample washing liquid of the sample washing zone 222 to drive the adsorption of the sample washing zone 222 by the driving device 3 to move the magnetic beads to be detected.
  • the sample to be detected is aggregated in the sample accumulation area 223;
  • the driving device 3 drives the magnetic microbeads into the sample signal amplifying pool 23, and the sample to be detected undergoes a polymerase chain reaction in the sample signal amplifying pool 23 to generate nucleic acid molecules, specifically, temperature regulation.
  • the device 4 performs a temperature cycle operation on the sample to be tested, and the operation includes the following steps: a, heating up to 94 ° C for 2 min; b, 94 ° C for 30 sec; c, cooling to 55 ° C for 30 sec; d, warming to 72 ° C for 30 sec; e, 72 ° C for 3 min; f. Cool down to 4 ° C for 5 min. Wherein step b to step e are cycled 20-25 times to generate sufficient nucleic acid molecules.
  • the detecting element detects the nucleic acid molecule.
  • the present invention provides a detection method for detecting a human defect immune virus, which sequentially drives a magnetic microbead with a sample to be detected into a sample adsorption zone 221, a sample washing zone 222, a sample aggregation zone 223, and a sample signal amplification cell 23 by means of a driving device 3.
  • the electrophoresis detector detects the sample to be detected after the signal amplification process is completed, and adopts an automatic operation to reduce the workload of the tester, and the reagent amount is small, and the reagent consumption is reduced, and the detection limit can reach the order of 1 fg, compared with
  • the existing conventional experimental method has a detection limit of 1 ng to improve the detection sensitivity.

Abstract

La présente invention concerne un dispositif d'essai du virus d'immunodéficience humaine (VIH) pour tester un échantillon d'essai, comprenant une pièce de support (1) et une puce de commande microfluidique (2) ; la puce de commande microfluidique (2) est pourvue d'un composant d'injection d'échantillon (21) pour placer les échantillons d'essai et des microbilles magnétiques, une zone de traitement d'échantillon (22) pour absorber, laver et collecter séquentiellement les échantillons d'essai, un composant d'amplification de signal (23) de piscine, un composant d'essai d'échantillon (24) en communication avec le composant d'amplification de signal d'échantillon (23), et des éléments d'essai disposés entre le composant d'amplification de signal d'échantillon (23) et le composant d'essai d'échantillon (24) ; le composant d'injection d'échantillon (21), la zone de traitement d'échantillon (22) et le composant d'amplification de signal d'échantillon (23) sont disposés séquentiellement dans un canal d'injection d'échantillon (25) et communiquent mutuellement ; la pièce de support (1) est pourvue d'un dispositif d'entraînement (3) sur celle-ci pour amener les microbilles magnétiques à entrer dans le composant d'amplification de signal d'échantillon (23) depuis le composant d'injection d'échantillon ; (21) et la pièce de support (1) est en outre pourvue d'un dispositif de réglage de température (4) pour ajuster la température de l'échantillon d'essai, et un dispositif de commande pour commander le dispositif d'entraînement (3) et le dispositif de réglage de température (4) et pour détecter le fonctionnement des éléments d'essai. Le dispositif a un degré élevé d'automatisation.
PCT/CN2014/072725 2014-02-28 2014-02-28 Dispositif d'essai du virus d'immunodéficience humaine et son procédé d'essai WO2015127656A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
PCT/CN2014/072725 WO2015127656A1 (fr) 2014-02-28 2014-02-28 Dispositif d'essai du virus d'immunodéficience humaine et son procédé d'essai

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN2014/072725 WO2015127656A1 (fr) 2014-02-28 2014-02-28 Dispositif d'essai du virus d'immunodéficience humaine et son procédé d'essai

Publications (1)

Publication Number Publication Date
WO2015127656A1 true WO2015127656A1 (fr) 2015-09-03

Family

ID=54008171

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2014/072725 WO2015127656A1 (fr) 2014-02-28 2014-02-28 Dispositif d'essai du virus d'immunodéficience humaine et son procédé d'essai

Country Status (1)

Country Link
WO (1) WO2015127656A1 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109456886A (zh) * 2018-12-12 2019-03-12 郑州科蒂亚生物技术有限公司 一种全自动核酸信号放大检测自动工作站
CN109499636A (zh) * 2019-01-14 2019-03-22 大连大学 一种病原体免疫检测数字微流控芯片及其制作方法
CN112955730A (zh) * 2019-03-06 2021-06-11 松下知识产权经营株式会社 病原体检测装置及病原体检测方法
CN112986554A (zh) * 2019-12-17 2021-06-18 中国农业大学 基于离心式微流控的牛奶中小分子检测方法及其专用芯片

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1791509A (zh) * 2002-12-30 2006-06-21 加州大学评议会 检测和分析病原体的方法和设备
CN101643701A (zh) * 2009-07-23 2010-02-10 清华大学 基于免疫磁性分离技术的细胞分选微流控芯片及其应用
CN102083533A (zh) * 2008-04-10 2011-06-01 芬兰技术研究中心 微流控芯片装置及其使用
CN102422143A (zh) * 2009-03-10 2012-04-18 新加坡科技研究局 处理生物样品和/或化学样品的装置
US20130065280A1 (en) * 2011-09-14 2013-03-14 Samsung Electronics Co., Ltd. Microfluidic apparatus and control method thereof
CN203825019U (zh) * 2014-02-28 2014-09-10 深圳市第二人民医院 人类免疫缺陷病毒检测装置
CN203825018U (zh) * 2014-02-28 2014-09-10 深圳市第二人民医院 人类免疫缺陷病毒检测装置
CN203825016U (zh) * 2014-02-28 2014-09-10 深圳市第二人民医院 人类免疫缺陷病毒检测装置

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1791509A (zh) * 2002-12-30 2006-06-21 加州大学评议会 检测和分析病原体的方法和设备
CN102083533A (zh) * 2008-04-10 2011-06-01 芬兰技术研究中心 微流控芯片装置及其使用
CN102422143A (zh) * 2009-03-10 2012-04-18 新加坡科技研究局 处理生物样品和/或化学样品的装置
CN101643701A (zh) * 2009-07-23 2010-02-10 清华大学 基于免疫磁性分离技术的细胞分选微流控芯片及其应用
US20130065280A1 (en) * 2011-09-14 2013-03-14 Samsung Electronics Co., Ltd. Microfluidic apparatus and control method thereof
CN203825019U (zh) * 2014-02-28 2014-09-10 深圳市第二人民医院 人类免疫缺陷病毒检测装置
CN203825018U (zh) * 2014-02-28 2014-09-10 深圳市第二人民医院 人类免疫缺陷病毒检测装置
CN203825016U (zh) * 2014-02-28 2014-09-10 深圳市第二人民医院 人类免疫缺陷病毒检测装置

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109456886A (zh) * 2018-12-12 2019-03-12 郑州科蒂亚生物技术有限公司 一种全自动核酸信号放大检测自动工作站
CN109499636A (zh) * 2019-01-14 2019-03-22 大连大学 一种病原体免疫检测数字微流控芯片及其制作方法
CN109499636B (zh) * 2019-01-14 2024-02-27 大连大学 一种病原体免疫检测数字微流控芯片及其制作方法
CN112955730A (zh) * 2019-03-06 2021-06-11 松下知识产权经营株式会社 病原体检测装置及病原体检测方法
CN112955730B (zh) * 2019-03-06 2024-04-19 松下知识产权经营株式会社 病原体检测装置及病原体检测方法
CN112986554A (zh) * 2019-12-17 2021-06-18 中国农业大学 基于离心式微流控的牛奶中小分子检测方法及其专用芯片
CN112986554B (zh) * 2019-12-17 2022-07-26 中国农业大学 基于离心式微流控的牛奶中小分子检测方法及其专用芯片

Similar Documents

Publication Publication Date Title
CN104374930B (zh) 人类免疫缺陷病毒检测装置及其检测方法
WO2015127656A1 (fr) Dispositif d'essai du virus d'immunodéficience humaine et son procédé d'essai
Lourbakos et al. Cleavage and activation of proteinase-activated receptor-2 on human neutrophils by gingipain-R from Porphyromonas gingivalis
Samloff et al. Electrophoretic heterogeneity and relationships of pepsinogens in human urine, serum, and gastric mucosa
DE60322729D1 (de) Zugängliche assay-einrichtung und verwendungsverfahren
DE69937368D1 (de) Nachweis von erkrankungen der nieren und behandlung
NO972611L (no) Fremgangsmåte for påvisning av nukleinsyrer med en bestemt sekvenssammensetning
EP3167045A1 (fr) Dispositif de réaction en chaîne par polymérase délocalisé pour le dépistage de maladies
DE69806024D1 (de) Erkennung und identifizierung von menschlichem papillomavirus durch pcr und typ-spezifischer reverser hybridisierung
ATE423217T1 (de) Assays zur überwachung von krebspatienten auf grundlage der spiegel von analytenkomponenten des plasminogen-aktivator-systems in proben von körperflüssigkeiten
KR20120020211A (ko) 치주 질환을 검출하기 위한 시험 키트
WO2019075868A1 (fr) Méthode de détection d'une infection par brucella et son application
PT1283907E (pt) Metodo para o diagnostico de cancros
Nunes et al. Taenia saginata: polymerase chain reaction for taeniasis diagnosis in human fecal samples
CN203825016U (zh) 人类免疫缺陷病毒检测装置
EP0325225A2 (fr) Concentrés de facteurs de coagulation II, VII, IX et/ou X, leur préparation et application et méthode pour la détermination du facteur de coagulation VII
Pessoa-e-Silva et al. Evaluation of urine for Leishmania infantum DNA detection by real-time quantitative PCR
Chen et al. Nanoparticle-based lateral flow biosensors integrated with loop-mediated isothermal amplification for the rapid and visual diagnosis of hepatitis B virus in clinical application
Rodríguez-Vega et al. Quantitative analysis of proteins secreted by Leishmania (Viannia) braziliensis strains associated to distinct clinical manifestations of American Tegumentary Leishmaniasis
Lichtinghagen et al. Identification of Bordetella pertussis in nasopharyngeal swabs using the polymerase chain reaction: evaluation of detection methods
Zhang et al. Single-cell analysis by intracellular immuno-reaction and capillary electrophoresis with laser-induced fluorescence detection
CN203825018U (zh) 人类免疫缺陷病毒检测装置
Carroll et al. Antibacterial peptide from normal rabbit serum. 1. Isolation from whole serum, activity, and microbicidal spectrum
Meier et al. Snake venom protein C activators
JP4298850B2 (ja) Ｃ型肝炎ウィルス（ｈｃｖ）検出方法

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 14883936

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

32PN Ep: public notification in the ep bulletin as address of the adressee cannot be established

Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC (EPO FORM 1205A DATED 12/01/2017)

122 Ep: pct application non-entry in european phase

Ref document number: 14883936

Country of ref document: EP

Kind code of ref document: A1