CN110075935A - The micro-fluidic cartridge of multiple determination and application method - Google Patents
The micro-fluidic cartridge of multiple determination and application method Download PDFInfo
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- CN110075935A CN110075935A CN201910329723.9A CN201910329723A CN110075935A CN 110075935 A CN110075935 A CN 110075935A CN 201910329723 A CN201910329723 A CN 201910329723A CN 110075935 A CN110075935 A CN 110075935A
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- cartridge
- liquid storage
- storage pipe
- sluice valve
- pedestal
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- 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
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- 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
Abstract
The invention discloses a kind of micro-fluidic cartridge of multiple determination and application methods, wherein, the cartridge includes: piston pump, liquid storage pipe, liquid storage pipe lid, cartridge pedestal, cover board, gasket, sluice valve and bottom plate, wherein, piston pump push-and-pull driving cartridge fluid, sluice valve rotation is logical to close runner connection, liquid storage pipe storing reagent sample, liquid storage pipe lid seals liquid storage pipe, cartridge pedestal support connection all parts, flowpath conduit and cavity on cover plate for sealing cartridge pedestal, gasket installs sealing between cartridge pedestal and sluice valve, bottom plate is by fixed building assembly sluice valve in cartridge pedestal.The cartridge is integrated with liquid of the fluid-operated pump with valve and needed for reacting, it is connect with the external world without flow path, and the detection and analysis of the multi objective automation of nucleic acid, albumen and biochemistry can be carried out to biological sample, operating process is simple, inspection result is abundant, has very high clinical value.
Description
Technical field
The present invention relates to vitro diagnostic techniques field, in particular to the micro-fluidic cartridge of a kind of multiple determination and application side
Method.
Background technique
Integrated automation analysis is the development trend in in-vitro diagnosis field.For the molecular diagnosis in in-vitro diagnosis, collection
Nucleic acid detection technique at automation is the development trend of molecular diagnostic techniques, and significance is not only to solve manpower and experiment
The problem of room occupied space, moreover it is possible to sample be avoided to bring potential product pollution etc. to ask the harm of operator and separate operations
Topic, can effectively improve the reliability of clinical diagnosis, reduce cost.For the immune and biochemical diagnosis in in-vitro diagnosis, entirely certainly
Dynamic multi objective joint-detection is developing direction, can not only be used manpower and material resources sparingly, and abundant face can be provided for doctor
Bed checking information, many index optimum organization can be improved the verification and measurement ratio of disease, and reduction is failed to pinpoint a disease in diagnosis and mistaken diagnosis.
Reagent type needed for the reaction of in-vitro diagnosis three basic forms of it is various, complex steps, needs complicated liquid
Manipulation and detection device match.Thus the instrument and equipment for in-vitro diagnosis detection generally requires more liquid control unit
Part is difficult to meet the area that medical infrastructure falls behind and the demand that scene is detected immediately so that equipment volume is larger.And sample
Product process flow tends not to flexibly change according to actual needs, and type and the volume range for handling sample are smaller.
Summary of the invention
The present invention is directed to solve at least some of the technical problems in related technologies.
For this purpose, an object of the present invention is to provide a kind of micro-fluidic cartridge of multiple determination, which being capable of complete or collected works
At automatic processing sample, preferably meets in-vitro diagnosis and examine needs.
It is another object of the present invention to the application methods for proposing a kind of micro-fluidic cartridge of multiple determination.
In order to achieve the above objectives, one aspect of the present invention embodiment proposes a kind of micro-fluidic cartridge of multiple determination, comprising:
Piston pump, the piston pump are connected with sluice valve by sealing joint, for driving cartridge fluid；Liquid storage pipe, for storing sample
Product, reagent and waste liquid, wherein the liquid storage pipe bottom end is equipped with through-hole, is connect with cartridge pedestal by top sealing joint；Liquid storage
Pipe lid, the liquid storage pipe lid and the liquid storage pipe are tightly connected, wherein the liquid storage pipe is covered with less than the liquid storage inside pipe wall
The venthole of diameter；Cartridge pedestal, the cartridge pedestal be equipped with the liquid storage pipe connecting interface, multiple determination reaction chamber and its
The sample channel and exhaust pipe of no interference, the through-hole communicated with the sluice valve, the sluice valve placing groove, the bottom plate are solid
Stationary interface, is fixed with the detection reagent of freeze-drying in the multiple determination reaction chamber, the through-hole circumferentially branch；Cover board, institute
It states cover board to be located above the cartridge pedestal, with cartridge pedestal multi objective reaction chamber and its sample channel and row described in adhering and sealing
Feed channel；Gasket, the gasket are located at cartridge pedestal sluice valve placing groove top surface；Sluice valve, the sluice valve position
Below the gasket, in the cartridge pedestal sluice valve placing groove, the sluice valve be equipped with the piston pump interface,
Fluid line, outlet, venthole and the card slot to connect with external instrument, the piston pump interface, the fluid line with it is described
Outlet communicates, and the venthole is through the sluice valve and separated by a distance with the fluid line and the outlet, passes through
The card slot that the sluice valve connects with external instrument is controlled, by the gating valve outlet and the liquid storage pipe and the multi objective
Detect reaction chamber sample channel gating；Bottom plate, the bottom plate pass through fixed structure below the sluice valve, with the cartridge pedestal
Part assembles cartridge.
The micro-fluidic cartridge of the multiple determination of the embodiment of the present invention can carry out nucleic acid, albumen and biochemistry to biological sample
Multi objective automation detection and analysis, operating process is simple, and inspection result is abundant, there is very high clinical value, and
Versatility greatly enhances, and can broadly meet the detection needs of different type clinical sample, has fully integrated brought
Advantage, detect stability and it is more efficient, cost is lower, user's detection not of the same race uses same set of hardware systems, favorably
In the acceptance for improving user, promote the universal of in-vitro diagnosis.
In addition, the micro-fluidic cartridge of multiple determination according to the above embodiment of the present invention can also have following additional skill
Art feature:
Further, in one embodiment of the invention, the piston pump includes push-and-pull piston rod and piston cylinder, with root
The up and down motion of the piston rod is controlled according to the control signal of external instrument.
Further, in one embodiment of the invention, the number of the liquid storage pipe is at least two.
Further, in one embodiment of the invention, the ventilation hole location of the liquid storage pipe lid is equipped with hydrophobic
Air film.
Further, in one embodiment of the invention, the number of the cartridge pedestal multiple determination reaction chamber is
At least one.
Further, in one embodiment of the invention, cartridge pedestal multiple determination reaction chamber bottom is equipped with
The notch of fixed pan substrate.
Further, in one embodiment of the invention, the cover board is transparent material, anti-to obtain biological detection
It answers.
Further, in one embodiment of the invention, the gasket and the cartridge pedestal sluice valve placing groove
Shape size is identical, and is equipped with the through-hole of the cartridge pedestal in the lead to the hole site of the cartridge pedestal.
Further, in one embodiment of the invention, the cartridge pedestal is transparent plastic, and the gasket is
Elastic polymer material.
In order to achieve the above objectives, another aspect of the present invention embodiment proposes a kind of answering for micro-fluidic cartridge of multiple determination
With method, using the micro-fluidic cartridge of multiple determination as described in above-described embodiment, wherein method includes: step S1, reacts institute
Reagent is needed to be stored in advance in the liquid storage pipe of cartridge, and after the liquid storage pipe is added in sample by user, by the cartridge
It places in mating control instrument；Step S2 controls instrument using outside, passes through the cooperation control of the piston pump and the sluice valve
System to be shifted, be mixed, batch operation to multistep fluid, and is controlled according to the target temperature control of detection reaction, magnetic bead,
Biological detection reaction needed for completing；Step S3 controls the external control instrument and reads multi objective reaction chamber described in cartridge
Reaction result.
The application method of the micro-fluidic cartridge of the multiple determination of the embodiment of the present invention, can to biological sample carry out nucleic acid,
The detection and analysis of albumen and the automation of biochemical multi objective, operating process is simple, and inspection result is abundant, and there is very high clinic to answer
With value, and versatility greatly enhances, and can broadly meet the detection needs of different type clinical sample, has fully integrated
Brought advantage, detect stability and it is more efficient, cost is lower, user's detection not of the same race uses same set of hardware item
System is conducive to the acceptance for improving user, promotes the universal of in-vitro diagnosis.
The additional aspect of the present invention and advantage will be set forth in part in the description, and will partially become from the following description
Obviously, or practice through the invention is recognized.
Detailed description of the invention
Above-mentioned and/or additional aspect and advantage of the invention will become from the following description of the accompanying drawings of embodiments
Obviously and it is readily appreciated that, in which:
Fig. 1 is the structural schematic diagram according to the micro-fluidic cartridge of multiple determination of the embodiment of the present invention；
Fig. 2 is the structural schematic diagram according to the micro-fluidic cartridge of multiple determination of first embodiment of the invention；
Fig. 3 is the structural schematic diagram according to the cartridge pedestal of first embodiment of the invention；
Fig. 4 is the structural schematic diagram according to the sluice valve of first embodiment of the invention；
Fig. 5 a and Fig. 5 b are sluice valve, the gating liquid storage tank cross-sectional view according to first embodiment of the invention；
Fig. 6 a to Fig. 6 o is the different phase sluice valve gating flow path signal that worked according to the cartridge of first embodiment of the invention
Figure；
Fig. 7 is to manipulate cartridge magnetic bead schematic diagram according to the electromagnet of first embodiment of the invention；
Fig. 8 a and Fig. 8 b are the liquid storage pipe cross-sectional view equipped with Silicon moulds according to second embodiment of the invention；
Fig. 9 is the structural schematic diagram according to the micro-fluidic cartridge of multiple determination of third embodiment of the invention；
Figure 10 is sluice valve, the gating flow path schematic diagram according to third embodiment of the invention；
Figure 11 is the structural schematic diagram according to the micro-fluidic cartridge of multiple determination of fourth embodiment of the invention；
Figure 12 is the application method flow chart according to the micro-fluidic cartridge of multiple determination of the embodiment of the present invention.
Specific embodiment
The embodiment of the present invention is described below in detail, examples of the embodiments are shown in the accompanying drawings, wherein from beginning to end
Same or similar label indicates same or similar element or element with the same or similar functions.Below with reference to attached
The embodiment of figure description is exemplary, it is intended to is used to explain the present invention, and is not considered as limiting the invention.
The micro-fluidic cartridge of the multiple determination proposed according to embodiments of the present invention and application method are described with reference to the accompanying drawings,
The micro-fluidic cartridge of multiple determination proposed according to embodiments of the present invention is described with reference to the accompanying drawings first.
Fig. 1 is the structural schematic diagram of the micro-fluidic cartridge of multiple determination of one embodiment of the invention.
As shown in Figure 1, the micro-fluidic cartridge of the multiple determination include: piston pump 100, liquid storage pipe 200, liquid storage pipe lid 300,
Cartridge pedestal 400, cover board 500, gasket 600 (specifically being marked not in Fig. 1), sluice valve 700 (specifically being marked not in Fig. 1)
With bottom plate 800.
Wherein, piston pump 100 is connected with sluice valve 700 by sealing joint, for driving cartridge fluid.Liquid storage pipe 200
For storing sample, reagent and waste liquid, wherein 200 bottom end of liquid storage pipe is equipped with through-hole, passes through top interface with cartridge pedestal 400
It is tightly connected.Liquid storage pipe lid 300 and liquid storage pipe 200 are tightly connected, wherein liquid storage pipe lid 300, which is equipped with, is less than 200 inner wall of liquid storage pipe
The venthole of diameter.Cartridge pedestal 400 be equipped with 200 connecting interface of liquid storage pipe, multiple determination reaction chamber and its without interference into
Sample pipeline and exhaust pipe, the through-hole communicated with sluice valve 700,700 placing groove of sluice valve, bottom plate fixed interface, multi objective inspection
It surveys in reaction chamber and is fixed with the detection reagent of freeze-drying, through-hole circumferentially branch.Cover board 500 is located at 400 top of cartridge pedestal, with viscous
Connect sealing 400 multi objective reaction chamber of cartridge pedestal and its sample channel and exhaust pipe.Gasket 600 be located at cartridge pedestal 400,
700 placing groove top surface of sluice valve.Sluice valve 700 is located at 600 lower section of gasket, in cartridge pedestal 400,700 placing groove of sluice valve
In, sluice valve 700 is equipped with and piston pump interface, fluid line, outlet, venthole and the card slot to connect with external instrument, piston
Pump 100 interfaces, fluid line is communicated with outlet, venthole is through sluice valve 700 and being separated by a spacing with fluid line and outlet
From the card slot to be connected by controlling sluice valve 700 with external instrument, by the outlet of sluice valve 700 and liquid storage pipe 200 and multi objective
Detect reaction chamber sample channel gating.Bottom plate 800 is assembled by fixing component with cartridge pedestal 400 and is blocked below sluice valve 700
Box.The cartridge of the embodiment of the present invention can carry out the detection point of the multi objective automation of nucleic acid, albumen and biochemistry to biological sample
Analysis, operating process is simple, and inspection result is abundant, has very high clinical value.
It should be noted that the micro-fluidic cartridge of multiple determination can in the application in biological detection or medical inspection, under
Face will be described in detail application of the cartridge in biological detection or medical inspection by embodiment.
In one embodiment of the invention, the cartridge of the embodiment of the present invention fully integrated nucleic acid field application, i.e.,
Nucleic acids in samples is extracted using based on paramagnetic particle method, and completes the nucleic acid fragment augmentation detection of multi objective.
Wherein, as shown in Figures 2 to 5, piston pump 100 includes push-and-pull piston rod 101 and piston cylinder 102, passes through external instrument
Device controls the up and down motion of piston rod 101, is connected with sluice valve 700 by the sealing of interface 103, drives cartridge fluid.Liquid storage pipe
200a to 200f is equipped with through-hole 201 for storing sample, reagent, waste liquid, bottom end, passes through top interface with cartridge pedestal 400
401a to 401f is tightly connected.Liquid storage pipe lid 300 and liquid storage pipe 200a to 200f are tightly connected, and are equipped with and are less than in liquid storage pipe 300
The venthole 301 of wall diameter is equipped with hydrophobic permeable membrane 302 at venthole 301.Cartridge pedestal 400 is equipped with liquid storage pipe connecting interface
401a to 401f, multiple determination reaction chamber 402 and its sample channel 403 and exhaust pipe 404 and sluice valve without interference
700 through-hole 405a to 405c, sluice valve placing groove 406, the bottom plate fixed interface 407a to 407c communicated, through-hole 405a to 405c
Circumferentially branch.Cover board 500 is located at 400 top of cartridge pedestal, seals cartridge pedestal multi objective reaction chamber 402 and its sample channel
403 and exhaust pipe 404.Gasket 600 is located at 406 top surface of cartridge pedestal sluice valve placing groove.Sluice valve 700 is located at gasket
600 lower section, in cartridge pedestal sluice valve placing groove 406, be equipped with piston pump interface 701, fluid line 702, outlet 703,
Venthole 704 and the card slot 705 to connect with external instrument, piston pump interface 701, fluid line 702 communicate with outlet 703, lead to
Stomata 704 is through sluice valve 700 and 703 separated by a distance with fluid line 702 and outlet, by control sluice valve 700 and
The card slot 705 that external instrument connects, by gating valve outlet 703 and liquid storage pipe 200 and multiple determination reaction chamber sample channel
403 gatings.Bottom plate 800 assembles cartridge by fixing component 801a to 801d below sluice valve 700, with cartridge pedestal 400.
Wherein, in one embodiment of the present of invention, cartridge pedestal 400 can be transparent plastic, and gasket 600 can be
Elastic polymer material.Certainly, cartridge pedestal 400 and gasket also can choose other kinds of material, herein only conduct
Example is not specifically limited.
Further, in one embodiment of the present of invention, the number of liquid storage pipe 200 is at least two, and cartridge pedestal refers to more
The number of mark detection reaction chamber 402 is at least one, and 200 He of liquid storage pipe can be arranged in those skilled in the art according to the actual situation
The specific number of cartridge pedestal multiple determination reaction chamber 402, is not specifically limited herein.
It should be noted that storing the reagent of nucleic acid extraction and amplification, multi objective into 200f in liquid storage pipe 200a in advance
The substrates such as preparatory immobilized primer in reaction chamber 402
Further, in one embodiment of the present of invention, cover board 500 can be transparent material, anti-to obtain biological detection
It answers.
It is understood that cover board 500 can be transparent material, compatible biological detection reaction, so that detection be facilitated to react
As a result.Certainly, cover board also can choose other kinds of material, herein only as an example, being not specifically limited.
Further, in one embodiment of the present of invention, gasket 600 and cartridge pedestal sluice valve place groove shape 406
Size is identical, and is equipped with the through-hole 601 of cartridge pedestal in the lead to the hole site of cartridge pedestal 400.
Further, the process for carrying out detection of nucleic acids using the cartridge of the present embodiment is as follows:
1) sample to be examined (such as blood, sputum, saliva, urine) is added in primary sample liquid storage pipe 200a, it is external
Instrument controlling sluice valve 700 gates liquid storage pipe 200a (shown in Fig. 6 a), at this point, gating valve outlet 703 and cartridge base plate through holes
405c is directed at (shown in Fig. 5 a-b), and 704 air-tight state of sluice valve through-hole, piston pump 100 extracts a certain amount of from liquid storage pipe 200a
Sample；
2) after completing sample extraction, sluice valve 700 rotates clockwise the gating second liquid storage pipe 200b that lysate is housed
(shown in Fig. 6 b).The quick pull of piston pump 100 several times, realizes being sufficiently mixed for sample and lysate；
3) after completing sample dissociation, sluice valve 700 rotates clockwise liquid storage pipe 200c (figure of the gating third equipped with magnetic bead
Shown in 6c).Several times, realization sample dissociation liquid is sufficiently mixed the quick pull of piston pump 100 with magnetic bead mixed liquor, completes sample core
After acid is adsorbed by magnetic bead, controls the electromagnet 900 below cartridge and open (shown in Fig. 7), piston pump 100 is slowly pushed, by waste liquid
It is pushed into third memotron 200c, magnetic bead is enriched to the underface of sluice valve 700；
4) after the enrichment for completing magnetic bead, sluice valve 700 rotates clockwise gating the 4th liquid storage pipe that cleaning solution is housed
200d (shown in Fig. 6 d), control cartridge below electromagnet 900 close, quick pull several times, realize the resuspension of magnetic bead and with it is clear
Washing lotion is sufficiently mixed, and after completing cleaning solution cleaning magnetic bead, is controlled the electromagnet 900 below cartridge and is opened, slowly push piston pump
100, waste liquid is pushed into the 4th memotron 200d, magnetic bead is enriched to the underface of sluice valve 700；
5) sluice valve 700 rotates clockwise gating the 5th liquid storage pipe 200e (shown in Fig. 6 e) that cleaning solution is housed, and repeats
(4) step carries out second to magnetic bead and cleans；
6) after the cleaning for completing magnetic bead, sluice valve 700 rotates clockwise gating the 6th liquid storage pipe that eluent is housed
200f (shown in Fig. 6 f) controls the electromagnet 900 below cartridge and closes, and the weight of magnetic bead is realized in quick pull piston pump 100 several times
It hangs and is sufficiently mixed with eluent；
6) it after completing Nucleic Acid Elution, controls the electromagnet 900 below cartridge and opens, nucleic acid solution is implanted sequentially and is referred to more
It marks in reaction chamber 402 (shown in Fig. 6 g-n), gates valve outlet 703 at this time and be aligned with cartridge base plate through holes 405a, sluice valve ventilation
Hole 704 is aligned with cartridge base plate through holes 405b；
7) after the completion of being loaded, cartridge sluice valve 700 is rotated, so that gating valve outlet 703 and venthole 704 are closed, is referred to more
It marks and realizes that physics is completely isolated (shown in Fig. 6 i) between reaction chamber 402, finally open the mating control instrument computer heating control in portion, realize more
Index amplification can use alternating temperature PCR amplification here, can also use constant-temperature amplification.Corresponding fluorescence detecting system can be real-time
The fluorescence signal value of Scanning Detction multi objective reaction chamber 402, interpretation testing result.
Above-mentioned elaborating for first embodiment of the invention, in second embodiment of the invention, such as Fig. 8 a-b institute
Show, the difference between this embodiment and the first embodiment lies in: the present embodiment uses the nucleic acid extraction scheme based on Silicon moulds, liquid storage pipe
Pellosil 1000 is fixed in the bottom 200c, and fluid control flow is different.
Further, the process for carrying out detection of nucleic acids using the cartridge of the present embodiment is as follows:
1) sample to be examined (such as blood, sputum, saliva, urine) is added in the liquid storage pipe 200a equipped with lysate,
External instrument controls sluice valve 700 and gates liquid storage pipe 200a (shown in Fig. 6 a), at this point, gating valve outlet 703 and cartridge pedestal
Through-hole 405c alignment, 704 air-tight state of sluice valve through-hole, the quick pull of piston pump 100 several times, realize filling for sample and lysate
Divide mixing；
2) after completing sample dissociation, sluice valve 700 rotates clockwise gating second liquid storage pipe that combination buffer is housed
200b (shown in Fig. 6 b).100 pull of piston pump several times, and is incubated for, and realizes being sufficiently mixed for sample dissociation liquid and combination buffer；
3) after, sluice valve 700 rotates clockwise liquid storage pipe 200c (Fig. 6 c that gating third is fixed with Silicon moulds 1000
It is shown), solution is pushed into liquid storage pipe 200c by piston pump 100, later slowly retracts the plunger up solution in pump 100, realizes silica gel
The enrichment of 1000 nucleic acid of mould captures；
4) after the enrichment for completing nucleic acid, rotation gates second liquid storage pipe 200b (shown in Fig. 6 b) to sluice valve 700 counterclockwise,
Waste liquid is discharged into second liquid storage pipe 200b by piston pump 100；
5) sluice valve 700 rotates clockwise gating the 4th liquid storage pipe 200d (shown in Fig. 6 d) that cleaning solution is housed, piston
Pump 100 extracts cleaning solution, and rotation gates third liquid storage pipe 200c (shown in Fig. 6 c) to sluice valve 700 counterclockwise later, slowly takes out
It draws several times, first time cleaning is carried out to Silicon moulds, the waste liquid of cleaning is pumped into piston pump 100, sluice valve 700 is rotated clockwise
The 4th liquid storage pipe 200d (shown in Fig. 6 d) is gated, piston pump 100 drains waste liquid；
6) sluice valve 700 rotates clockwise gating the 5th liquid storage pipe 200e (shown in Fig. 6 e) that cleaning solution is housed, and repeats
(5) step carries out second to magnetic bead and cleans, and waste liquid is discharged into liquid storage pipe 200e；
7) after completing the cleaning of two step of nucleic acid, sluice valve 700 rotates clockwise gating the 6th liquid storage pipe that eluent is housed
200f (shown in Fig. 6 f), piston pump 100 extract eluent, rotate sluice valve 700 counterclockwise and gate third liquid storage pipe 200c (figure
Shown in 6c), quick pull piston pump 100 realizes the elution of nucleic acid；
8) piston pump 100 extracts nucleic acid eluents, and assigns it in multi objective reaction chamber 402, carries out multi objective
Amplification Analysis detection.
In third embodiment of the invention, such as Fig. 9, shown in 10, the difference between this embodiment and the first embodiment lies in: this reality
It applies example and cartridge is used for protein immunization detection field, 402 bottom of multiple determination reaction chamber of cartridge pedestal 400 is equipped with fixed flat
The notch 408 of face substrate, ankyrin dot matrix chip 1100, fluid control flow are different.
Further, ankyrin dot matrix chip 1100 be preferably glass material, through agarose or it is epoxy group modified after,
Point sample is coated with primary antibody solution, and capture antibody used is fixed on Protein microdot arrays chip surface, carries out BSA later and closes core
Piece.
Further, the Protein microdot arrays chip 1100 that preparation is completed is fixedly mounted on cartridge pedestal fixed pan substrate
In notch 408.
Further, the process for carrying out multi-index protein detection using the cartridge of the present embodiment is as follows:
1) sample to be examined (such as serum) is added in first sample liquid storage pipe 200a, external instrument controls sluice valve
700 gating liquid storage pipe 200a, piston pump 100 extract a certain amount of sample from liquid storage pipe 200a, later 700 up time of sluice valve
Needle rotation gating multi objective reaction chamber 402, gates valve outlet 703 at this time and is aligned with cartridge base plate through holes 405a, sluice valve ventilation
Hole 704 is aligned with cartridge base plate through holes 405b, and 100 pull of piston pump several times, realizes the antigen and Protein microdot arrays chip in sample
Coated primary antibody sufficiently combines on 1100, drains the liquid in multi objective 402 chambers of reaction later, rotates clockwise sluice valve 700
Liquid storage pipe 200a is gated, liquid is pushed completely into liquid storage pipe 200a by piston pump 100 later；
2) sluice valve 700 rotates clockwise the gating second liquid storage pipe 200b that cleaning solution is housed, and piston pump 100 will be from storage
A certain amount of cleaning solution is extracted in liquid pipe 200b, sluice valve 700 rotates clockwise gating multi objective reaction chamber 402, piston later
It pumps 100 pulls several times, realizes the abundant cleaning of Protein microdot arrays chip 1100, drain the liquid in multi objective 402 chambers of reaction later,
It rotates clockwise sluice valve 700 and gates liquid storage pipe 200b, liquid is pushed completely into liquid storage pipe 200b by piston pump 100 later；
3) sluice valve 700 rotates clockwise the liquid storage pipe 200c that gating third is equipped with primary antibody, and piston pump 100 will be from liquid storage
A certain amount of primary antibody solution is extracted in pipe 200c, sluice valve 700 rotates clockwise gating multi objective reaction chamber 402, piston later
It pumps 100 pulls several times, realizes the abundant combination of 1100 primary antibody of Protein microdot arrays chip, drain multi objective later and react in 402 chambers
Liquid rotates clockwise sluice valve 700 and gates liquid storage pipe 200c, and liquid is pushed completely into liquid storage pipe 200c by piston pump 100 later
In；
4) sluice valve 700 rotates clockwise gating the 4th liquid storage pipe 200d that cleaning solution is housed, repetitive process (2)；
5) sluice valve 700 rotates clockwise the 5th liquid storage pipe 200e equipped with signal label secondary antibody of gating, piston pump 100
A certain amount of two corresponding anti-solution will be extracted from liquid storage pipe 200e, sluice valve 700 rotates clockwise gating multi objective reaction chamber later
402,100 pull of piston pump several times, realizes the abundant combination of 1100 secondary antibody of Protein microdot arrays chip, drains multi objective reaction later
Liquid in 402 chambers rotates clockwise sluice valve 700 and gates liquid storage pipe 200e, and liquid is pushed completely into storage by piston pump 100 later
In liquid pipe 200c；
6) sluice valve 700 rotates clockwise gating the 6th liquid storage pipe 200f that cleaning solution is housed, repetitive process (2)；
7) detection of 1100 signal of Protein microdot arrays chip is carried out, interpretation analysis as a result.
Furthermore similarly, the technical solution of the present embodiment can be used for the detection of nucleic acid hybridization hybrid chip.
In four embodiment of the invention, as shown in figure 11, the difference between this embodiment and the first embodiment lies in, this implementation
Cartridge is used for biochemistry detection field by example, need to only carry out simple dilute sample step, so that it may directly in multiple determination chamber 402
Carry out the detection of Multiple biomarkers.
Further, the biochemical reaction substrate of multi objective reaction chamber is blood parameters and the detection examination of different freeze-dryings
Agent, including glutamic-pyruvic transaminase, glutamic-oxalacetic transaminease, glutamyl transpeptidase, alkaline phosphatase, cholinesterase, phenylalanine ammonia group-transfer
Enzyme, aspartate amino transferase, γ-Gu ammonia phthalidyl transferase, total protein, albumin, glycosylated albumin, total bilirubin,
Bilirubin direct, ammonia, urea, creatinine, uric acid, glucose, total cholesterol, triglycerides, high-density lipoprotein, low-density lipoprotein
White, very low density lipoprotein, Apolipoprotein A1, apolipoprotein B, calcium, chlorine, iron, phosphorus, sodium, potassium, carbon dioxide.
Further, the process for carrying out multi objective biochemistry detection using the cartridge of the present embodiment is as follows:
1) sample to be examined (such as serum, blood plasma) is added in first sample liquid storage pipe 200a, external instrument control
Sluice valve 700 gates liquid storage pipe 200a, and piston pump 100 extracts quantitative sample from liquid storage pipe 200a, later sluice valve 700
Gating liquid storage pipe 200c is rotated clockwise, sample is pushed completely into liquid storage pipe 200c by piston pump 100 at this time；
2) rotation gates second liquid storage pipe 200b that dilution is housed to sluice valve 700 counterclockwise, and piston pump 100 will be from storage
Quantitative cleaning solution is extracted in liquid pipe 200b, sluice valve 700 rotates clockwise gating liquid storage pipe 200c later, and piston pump 100 is taken out
It draws several times, realizes being sufficiently mixed for sample and dilution；
3) piston pump 100 quantitatively extracts solution from liquid storage pipe 200c, and sluice valve 700 rotates clockwise choosing later, passes through
Solution is implanted sequentially in multi objective reaction chamber 402 by the precise match of piston pump 100 and sluice valve 700, different sample channels
403 and exhaust pipe 404 it is mutually isolated, prevented the possibility of cross contamination.
4) signal detection of multi objective reaction chamber 402 is carried out, interpretation analysis as a result.
The micro-fluidic cartridge of the multiple determination proposed according to embodiments of the present invention can carry out nucleic acid, egg to biological sample
The detection and analysis of white and biochemical multi objective automation, operating process is simple, and inspection result is abundant, has very high clinical application
Value, and versatility greatly enhances, and can broadly meet the detection needs of different type clinical sample, has fully integrated institute
The advantages of bringing, detect stability and it is more efficient, cost is lower, user's detection not of the same race uses same set of hardware item
System is conducive to the acceptance for improving user, promotes the universal of in-vitro diagnosis.
Referring next to the application method for the micro-fluidic cartridge of multiple determination that attached drawing description proposes according to embodiments of the present invention.
Figure 12 is the flow chart of the application method of the micro-fluidic cartridge of multiple determination of one embodiment of the invention.
As shown in figure 12, the application method of the micro-fluidic cartridge of the multiple determination is examined using the multi objective of above-described embodiment
Micrometer flow control cartridge, wherein method includes:
Step S1, reagent needed for reacting is stored in advance in the liquid storage pipe of cartridge, and liquid storage pipe is added in sample in user
Afterwards, cartridge is placed in mating control instrument.
Step S2 controls instrument using outside, is controlled by the cooperation of piston pump and sluice valve, to carry out to multistep fluid
Transfer, mixing, batch operation, and controlled according to the target temperature control of detection reaction, magnetic bead, it is anti-to complete required biological detection
It answers.
Step S3 controls the reaction result that external control instrument reads cartridge multi objective reaction chamber.
It should be noted that the aforementioned explanation to the micro-fluidic cartridge embodiment of multiple determination is also applied for the implementation
The application method of the micro-fluidic cartridge of multiple determination of example, details are not described herein again.
The application method of the micro-fluidic cartridge of the multiple determination proposed according to embodiments of the present invention, can to biological sample into
The detection and analysis of the multi objective automation of row nucleic acid, albumen and biochemistry, operating process is simple, and inspection result is abundant, has very high
Clinical value, and versatility greatly enhances, and can broadly meet the detection needs of different type clinical sample, tool
Have it is fully integrated brought by advantage, detect stability and it is more efficient, cost is lower, user's detection not of the same race uses same
Hardware systems are covered, the acceptance for improving user is conducive to, promote the universal of in-vitro diagnosis.
In addition, term " first ", " second " are used for descriptive purposes only and cannot be understood as indicating or suggesting relative importance
Or implicitly indicate the quantity of indicated technical characteristic.Define " first " as a result, the feature of " second " can be expressed or
Implicitly include at least one this feature.In the description of the present invention, the meaning of " plurality " is at least two, such as two, three
It is a etc., unless otherwise specifically defined.
In the description of this specification, reference term " one embodiment ", " some embodiments ", " example ", " specifically show
The description of example " or " some examples " etc. means specific features, structure, material or spy described in conjunction with this embodiment or example
Point is included at least one embodiment or example of the invention.In the present specification, schematic expression of the above terms are not
It must be directed to identical embodiment or example.Moreover, particular features, structures, materials, or characteristics described can be in office
It can be combined in any suitable manner in one or more embodiment or examples.In addition, without conflicting with each other, the skill of this field
Art personnel can tie the feature of different embodiments or examples described in this specification and different embodiments or examples
It closes and combines.
Although the embodiments of the present invention has been shown and described above, it is to be understood that above-described embodiment is example
Property, it is not considered as limiting the invention, those skilled in the art within the scope of the invention can be to above-mentioned
Embodiment is changed, modifies, replacement and variant.
Claims (10)
1. a kind of micro-fluidic cartridge of multiple determination characterized by comprising
Piston pump, the piston pump are connected with sluice valve by sealing joint, for driving cartridge fluid；
Liquid storage pipe, for storing sample, reagent and waste liquid, wherein the liquid storage pipe bottom end is equipped with through-hole, passes through with cartridge pedestal
The connection of top sealing joint；
Liquid storage pipe lid, the liquid storage pipe lid and the liquid storage pipe are tightly connected, wherein the liquid storage pipe is covered with less than the storage
The venthole of liquid pipe inner diameter；
Cartridge pedestal, the cartridge pedestal are equipped with the liquid storage pipe connecting interface, multiple determination reaction chamber and its without interference
Sample channel and exhaust pipe, the through-hole communicated with the sluice valve, the sluice valve placing groove, the bottom plate fixed interface,
It is fixed with the detection reagent of freeze-drying in the multiple determination reaction chamber, the through-hole circumferentially branch；
Cover board, the cover board are located above the cartridge pedestal, with cartridge pedestal multi objective reaction chamber described in adhering and sealing and its
Sample channel and exhaust pipe；
Gasket, the gasket are located at the cartridge pedestal, sluice valve placing groove top surface；
Sluice valve, the sluice valve is located at below the gasket, in the cartridge pedestal, sluice valve placing groove, the choosing
Port valve is equipped with and the piston pump interface, fluid line, outlet, venthole and the card slot to connect with external instrument, the piston
Pump interface, the fluid line with it is described outlet communicate, the venthole through the sluice valve and with the fluid line and
The outlet is separated by a distance, the card slot to be connected by controlling the sluice valve with external instrument, by the gating valve outlet
It is gated with the liquid storage pipe and the multiple determination reaction chamber sample channel；And
Bottom plate, the bottom plate assemble cartridge by fixing component below the sluice valve, with the cartridge pedestal.
2. the micro-fluidic cartridge of multiple determination according to claim 1, which is characterized in that the piston pump includes that push-and-pull is lived
Stopper rod and piston cylinder control the up and down motion of the piston rod with the control signal according to external instrument.
3. the micro-fluidic cartridge of multiple determination according to claim 1, which is characterized in that the number of the liquid storage pipe is extremely
It is two few.
4. the micro-fluidic cartridge of multiple determination according to claim 1, which is characterized in that the venthole of the liquid storage pipe lid
Hydrophobic permeable membrane is equipped at position.
5. the micro-fluidic cartridge of multiple determination according to claim 1, which is characterized in that the cartridge pedestal multi objective inspection
The number for surveying reaction chamber is at least one.
6. the micro-fluidic cartridge of multiple determination according to claim 1, which is characterized in that the cartridge pedestal multi objective inspection
Survey the notch that reaction chamber bottom is equipped with fixed pan substrate.
7. the micro-fluidic cartridge of multiple determination according to claim 1, which is characterized in that the cover board is transparent material,
To obtain biological detection reaction.
8. the micro-fluidic cartridge of multiple determination according to claim 1, which is characterized in that the gasket and the cartridge
Pedestal sluice valve placing groove shape size is identical, and is equipped with the logical of the cartridge pedestal in the lead to the hole site of the cartridge pedestal
Hole.
9. the micro-fluidic cartridge of multiple determination according to claim 1, which is characterized in that the cartridge pedestal is transparent modeling
Material, and the gasket is elastic polymer material.
10. a kind of application method of the micro-fluidic cartridge of multiple determination, which is characterized in that using such as any one of claim 1-9
The micro-fluidic cartridge of the multiple determination, wherein method includes:
Step S1, reagent needed for reacting is stored in advance in the liquid storage pipe of cartridge, and the storage is added in sample in user
After liquid pipe, the cartridge is placed in mating control instrument；
Step S2 controls instrument using outside, is controlled by the cooperation of the piston pump and the sluice valve, to multistep fluid
It shifted, mixed, batch operation, and being controlled according to the target temperature control of detection reaction, magnetic bead, completing required biology inspection
Survey reaction；
Step S3 controls the reaction result that the external control instrument reads multi objective reaction chamber described in cartridge.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910329723.9A CN110075935B (en) | 2019-04-23 | 2019-04-23 | Multi-index detection microfluidic card box and application method |
| PCT/CN2020/083549 WO2020216058A1 (en) | 2019-04-23 | 2020-04-07 | Multi-index test microfluidic kit and application method therefor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910329723.9A CN110075935B (en) | 2019-04-23 | 2019-04-23 | Multi-index detection microfluidic card box and application method |
Publications (2)
| Publication Number | Publication Date |
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| CN110075935A true CN110075935A (en) | 2019-08-02 |
| CN110075935B CN110075935B (en) | 2020-06-19 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910329723.9A Active CN110075935B (en) | 2019-04-23 | 2019-04-23 | Multi-index detection microfluidic card box and application method |
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| Country | Link |
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| CN (1) | CN110075935B (en) |
| WO (1) | WO2020216058A1 (en) |
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| CN110711612A (en) * | 2019-10-28 | 2020-01-21 | 杭州欣创医疗器械有限公司 | Portable recyclable tissue sample inspection bottle and filling device thereof |
| CN110964716A (en) * | 2019-12-05 | 2020-04-07 | 广州万孚生物技术股份有限公司 | In-vitro diagnosis and analysis device and reagent card |
| CN111394221A (en) * | 2020-04-14 | 2020-07-10 | 无锡科智达科技有限公司 | Totally-enclosed multi-index nucleic acid detection device |
| WO2020216058A1 (en) * | 2019-04-23 | 2020-10-29 | 清华大学 | Multi-index test microfluidic kit and application method therefor |
| CN112226358A (en) * | 2020-12-11 | 2021-01-15 | 博奥生物集团有限公司 | Nucleic acid analysis card box and nucleic acid analysis equipment |
| CN113025489A (en) * | 2021-05-28 | 2021-06-25 | 博奥生物集团有限公司 | Fully integrated nucleic acid analysis system |
| CN113383221A (en) * | 2019-09-30 | 2021-09-10 | 伊鲁米纳新加坡有限公司 | Test box of analytical instrument |
| CN113667598A (en) * | 2021-08-17 | 2021-11-19 | 圣湘生物科技股份有限公司 | Nucleic acid detection device and nucleic acid detection method |
| CN114308149A (en) * | 2021-11-29 | 2022-04-12 | 北京机械设备研究所 | Chip sealing device and self-sealing modular chip equipment |
| CN114517151A (en) * | 2022-02-16 | 2022-05-20 | 东南大学 | Liquid control component and conversion combination use method thereof |
| CN115382586A (en) * | 2021-05-24 | 2022-11-25 | 广州万孚生物技术股份有限公司 | Driving device and medical detection system |
| WO2023216184A1 (en) * | 2022-05-12 | 2023-11-16 | 广州国家实验室 | Cartridge for nucleic acid amplification |
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| CN114308149A (en) * | 2021-11-29 | 2022-04-12 | 北京机械设备研究所 | Chip sealing device and self-sealing modular chip equipment |
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Also Published As
| Publication number | Publication date |
|---|---|
| CN110075935B (en) | 2020-06-19 |
| WO2020216058A1 (en) | 2020-10-29 |
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