CN104897654B - A kind of microflow controlled biochip detection device and preparation method - Google Patents
A kind of microflow controlled biochip detection device and preparation method Download PDFInfo
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- CN104897654B CN104897654B CN201510294370.5A CN201510294370A CN104897654B CN 104897654 B CN104897654 B CN 104897654B CN 201510294370 A CN201510294370 A CN 201510294370A CN 104897654 B CN104897654 B CN 104897654B
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Abstract
The invention discloses a kind of microflow controlled biochip detection device and preparation methods.The detection device is mainly made of grooved drum plate, bottom plate, groove and notch, and test point is equipped in the groove.The detection device is prepared, using simple, and suitable for mass producing, applying, and testing result is reliable and stable.
Description
Technical field
The present invention relates to a kind of microflow controlled biochip detection device and preparation methods.
Background technology
In-vitro diagnosis product technology mainly has Western blotting and enzyme-linked immunologic diagnosis technology (ELISA) currently on the market, leads to
The presence for interacting to detect corresponding antigens or antibody between antigen and antibody is crossed, so as to reach the mesh for diagnosing certain disease
's.
Western blot is a kind of protein identification techniques being widely used in life science, test need by
Electrophoresis, transfer and detection and etc., operating procedure is complicated, and can only be polluted every time with a kind of serum, and interference is surveyed
Test result, the device are susceptible to the result of false negative.
ELISA is most widely used in immunoassay, and antigen or antibody are measured by the use of orifice plate as solid phase carrier
A kind of technology.ELISA kit detects the diagnosis for having been used for a variety of diseases, in practice it has proved that is effective.But mesh
The preceding ELISA kit sold in the market is all largely the detection of single index, and detection time is long, costly, to patient with
Carry out heavier financial burden；Further, since parallel sample detection delay and experimental result is caused difference occur, give diagnosis increase
Difficulty extends Diagnostic Time, and puzzlement is brought to doctor and patient, affects early stage medical diagnosis on disease and treatment.
Both the above product testing cost is higher, and time-consuming, and the integrality of information and the accuracy of result cannot be definite
Guarantee.And be frequently necessary to be detected the different material in a variety of samples in actual life, the complete of sample could be obtained
Information, it is necessary to quickly and accurately obtain these information, experimental error could be reduced, ensure the confidence level of data.
Patent CN 101526520B disclose a kind of detection method and device of biological sample, although can realize multiple
The detection of many indexes of sample, however its biomolecule is coated on bottom plate, the first tablet when either sample preparation still detects
Relative movement cannot be generated with the second tablet, otherwise can destroy the biomolecule on the second tablet, it is therefore, tight to requirement of experiment
Lattice, so as to also be difficult to guarantee detection quality.In addition, when carrying out sample preparation and detection using its device, since its first tablet is had to
, it is necessary to which manual shift position, causes its sample preparation and test position to be difficult to be consistent in activity, so that difficult during detection
Instrument to be used accurately to be read to testing result automatically.Further, since its first tablet is had in activity, it can not
It is fixed so that its application scenario is limited, and is suitable only for the temporary detecting of a small amount of sample in laboratory, can not industrial mass manufacture answer
With；Simultaneously as its first tablet needs are in active state, it is difficult to which realization is combined with the sealing of the second tablet, causes its system
The problem of being difficult to eliminate sample liquid cross influence (pollution) when sample or detection influences the stability of testing result；And due to its
One tablet needs are in active state, it is difficult to which avoiding outer bound pair, it prepares the influence of sample, therefore testing conditions requirement is strictly, simultaneously
And requirement is rapidly completed in a short time from sample preparation to detection, to reduce the influence of outer bound pair testing result stability.
Against the above deficiency, it would be highly desirable to develop a kind of detection method easy to operate, testing result is reliable and stable and device.
The content of the invention
It is an object of the invention to provide a kind of microflow controlled biochip detections easy to operate, testing result is reliable and stable
Device and preparation method thereof.
To achieve the above object, concrete scheme of the invention is as follows：
The present invention provides a kind of microflow controlled biochip detection device, and the detection device includes grooved drum plate, bottom plate；It is described
Grooved drum plate includes first surface and the second surface opposite with first surface, and close by the first surface and the bottom plate
Sealed knot closes；The first surface of the grooved drum plate be equipped at least one Uncrossed groove, every groove with the grooved drum plate
First surface border between leave distance, and at least one test point is equipped in every groove, on each test point point
Fu Zai there be a kind of capture molecule；The second surface of the grooved drum plate is equipped with notch, and the notch is arranged at described second
With the corresponding position in every groove both ends on surface, and connected with the groove, it is to be tested for being added into every groove
Sample.
In the present invention, the sealing is with reference to referring between the first surface and the bottom plate in close and groove
Substance (sample) is difficult to the combination that interface therebetween is retained, and avoids the cross influence of substance between multiple grooves.
In one preferred embodiments, pass through thermal bonding, ultrasonic bond or the side of being mechanically fixed between the first surface and the bottom plate
Formula realizes that sealing combines.
In the present invention, combined to be conducive to the sealing between the first surface and the bottom plate, it is preferable that described
Grooved drum plate and bottom plate are simultaneously tablet or are the plate with the radian that matches.
Detection device according to the present invention, it is preferable that in every groove be equipped at least two test points, described at least two
Load has different capture molecules respectively on a test point, so as to detect multiple indexs simultaneously.Preferably, the capture
Antigen, antibody or probe are selected from molecule.
Detection device according to the present invention, it is preferable that the width of the groove is 100-5000 μm, the depth of the groove
For 100-5000 μm；The test point is located at the bottom of the groove.
In the present invention, the test point is made by that can adsorb capture of the material of molecule, for example, selected from simple glass,
Polystyrene, polyvinyl chloride, makrolon, organic glass, dimethyl silicone polymer or they are modified through active group
Material.In one embodiment, the location point that the test point is formed by the region for directly forming the trench bottom surfaces,
Capture at this time is directly loaded in molecule on the bottom surface of the groove；In another embodiment, the test point is by it
The location point that the object that he is attached in the trench bottom surfaces is formed, such as the test point can be by colored material
Attachment precalculated position in the trench is formed, consequently facilitating point sample and being conducive to the selection of groove plate material (such as selection being dredged
Water material), be conducive to the cleaning of sample liquid during subsequent detection, wherein the adhering mode of the object in the trench can there are many,
To be known in the art, which is not described herein again.
In one embodiment of the invention, the first surface is equipped with a Uncrossed groove.In the present invention
Another embodiment in, the first surface be equipped at least two Uncrossed grooves, so as to simultaneously to multiple
Multiple indexs of sample are detected.Preferably, the groove is S-type or snakelike.
In the present invention, the notch is used to add sample to be tested into every groove, it is preferable that the notch is in cone
Shape, tapered bottom are connected with the groove, are more advantageous to being loaded and be cleared up；Preferably, the notch is located at described the
Upper end diameter on two surfaces is 1.5-5mm, further preferably 2-4mm, more preferably 2.5-3.5mm.
Detection device according to the present invention, it is preferable that sealant is additionally provided on the second surface, for closing
Notch is stated, since the equal concentrated setting of notch is on second surface, sealing is easy.
The present invention also provides a kind of preparation methods of above-mentioned detection device, include the following steps：
A, at least one groove is formed on the first surface of grooved drum plate, and it is on the second surface of the grooved drum plate and every
The corresponding position in groove both ends opens up notch, and the notch is connected with the groove, to be measured in order to be added into groove
Test agent；
At least one capture is put to precalculated position in the trench respectively with molecule by point sample mode, to be formed at least b,
One load has a kind of test point of capture molecule；
C, the first surface of the bottom plate and the grooved drum plate is sealed and combined, obtain the detection device.
The present invention also provides it is a kind of using above-mentioned detection device carry out biological detection method, including：
The first step：Detected sample is injected into the groove by the notch of described groove one end；
Second step：The detected sample and the capture being supported on the test point are carried out with the interaction of molecule
Detection.In one embodiment, the second step is to the detected sample and the capture being supported on the test point
It is developed the color or is shone with the interaction of molecule and detected；It is further preferred that in the second step, directly pass through instrument pair
The colour developing of test point or luminous result are read.
Compared with prior art, the present invention has the following advantages：
1st, detection device of the invention prepares the biomolecule such as antigen or antibody in the trench using point sample mode, with biography
The tablet coating sample preparation of system is compared, and preparation process is simple, and since biomolecule is prepared in the trench, grooved drum plate may be used also
It is either when subsequently sealing combination with bottom plate or follow-up in use, point sample system to play certain protective effect to sample
Standby biomolecule detection point will not be mechanically damaged, therefore quality is more stable.
2nd, detection device of the invention in the preparation, the sample precalculated position of point in the trench by way of point sample, with
When preparing sample on tablet, sample detection position is difficult to prepare to ensure to compare in the prior art, not only contributes to big batch
Change, standardized production is conducive to directly by instrument in subsequent detection to precalculated position on grooved drum plate with reducing cost
The detection information of (test point) is read automatically, so as to ensure read detection information while work efficiency is improved
Reliability.
3rd, in detection device of the invention biomolecule point in the trench, not with contacts baseplate, by bottom plate grooved drum plate with
It can bear fiercer condition during bonding (such as thermal bonding, the ultrasonic bond, be mechanically fixed when) of bottom plate, therefore, follow-up
Will not be impacted in being bonded of grooved drum plate and bottom plate, production technology is simpler, is more easy to accomplish scale production.
4th, in detection device of the invention, due to leaving distance between the border of groove and the first surface of the grooved drum plate
(i.e. described groove does not intersect with above-mentioned border, such as the distance not less than 1mm), along with the sealing of the bottom plate combines,
So that the groove is only in communication with the outside by the notch, therefore, detection device of the invention is being produced, pack, deposited
During storage and use, it can lack by ectocine；The detection device of the present invention can also be further in the second surface
Be simply to set sealant (such as sealing film) can sealing groove mouth, so as to fulfill the reliable sealing of groove, to ensure biology
The stability of molecule sample preparation, and then improve the reliability of testing result.
5th, detection device of the invention notes sample and washing lotion when being detected from notch, simple and fast, easily quantifies, and
And due to noting sample from notch, extra sample liquid can be converged at the notch of the groove other end, is convenient to clean, when grooved drum plate is equipped with
Cross influence is not susceptible to during a plurality of groove；And since bottom plate and grooved drum plate by thermal bonding, ultrasonic bond or are mechanically fixed
Etc. means sealing combine, when detecting it is possible to prevente effectively from sample liquid cross influence between grooved drum plate and bottom plate；Simultaneously as make
Used time operating procedure is simple and fast, easily-controllable, therefore testing result is more stable, convenient for large-scale promotion application.
6th, there are different antigen, antibody or probe in the groove of detection device of the invention, a variety of samples can be obtained simultaneously
This multiple indexs, i.e., detect simultaneously so that experiment is completed under the same conditions as far as possible, substantially increases the accuracy of experiment；
Meanwhile apparatus of the present invention can be produced in batches, cost is relatively low, reduces the financial burden of patient, and large-scale promote is suitble to make
With.
Description of the drawings
The point sample schematic diagram of Fig. 1 microflow controlled biochip detection devices；
The sealing-in schematic diagram of Fig. 2 microflow controlled biochip detection devices；
The microflow controlled biochip detection device schematic diagram of Fig. 3 present invention；
The testing result of Fig. 4 embodiments 5；
The testing result of Fig. 5 embodiments 6；
The testing result of Fig. 6 embodiments 7；
The testing result of Fig. 7 embodiments 8.
Specific embodiment
Below with reference to attached drawing, the present invention will be described in more detail.
Fig. 1-3 gives microflow controlled biochip detection device of the invention and its preparation, using process.The present invention's is micro-
Micro-continuous-flow biochip detection device includes grooved drum plate 1, bottom plate 2；The grooved drum plate 1 include first surface 11 and with first surface 11
Opposite second surface 12, and combined by the first surface 11 and the bottom plate 2 sealing；The first of the grooved drum plate 1
Surface 11 is equipped at least one Uncrossed groove 3, border of the every groove 3 with the first surface 11 of the grooved drum plate 1
Between leave distance, and at least one test point (not shown) is equipped in every groove 3, is born respectively on each test point
It is loaded with a kind of capture molecule；The second surface 12 of the grooved drum plate 1 is equipped with notch 4, and the notch 4 is arranged at described second
It with the corresponding position in every 3 both ends of groove on surface 12, and connects with the groove 3, is treated for being added into every groove 3
Test sample.
In a preferred embodiment, the notch 4 is tapered, and tapered bottom is connected with the groove 3, is more had
Beneficial to sample-adding and cleaning；Preferably, the notch 3 is located at the upper end diameter on the second surface 12 for 1.5-5mm, further
Preferably 2-4mm, more preferably 2.5-3.5mm.In one embodiment, sealant is additionally provided on the second surface 12, used
In closing the notch 4, since 4 equal concentrated setting of notch is on second surface 12, sealing is easy.
It is specific when preparing, form at least one groove 3 on grooved drum plate 1, capture molecule can be existed by 5 points of sample applicator
Form test point in groove 3,2 sealing-in of excellent grooved drum plate 1 and bottom plate (sealing combine) will be put, so as to groove 3 and bottom plate 2 it
Between form pipeline, so as to be combined into flexible biological probe.In use, sample and various is added at the device notch 4
Detection reagent is detected.
In the apparatus of the present, the grooved drum plate 1 and bottom plate 2 can be for tablets or with the curved of the radian that matches
Curved shape.The material of grooved drum plate 1 and bottom plate 2 can be organic material or glass etc., for example, selected from simple glass, polystyrene,
Polyvinyl chloride, makrolon, organic glass, dimethyl silicone polymer and they through the modified material of active group.
In addition, in order to further improve the closure of pipeline, it is also contemplated that set between grooved drum plate 1 and bottom plate 2 to
Few one layer of mantle, the material of the mantle is mainly PDMS (dimethyl silicone polymer) material, by PDMS and curing agent according to 10：
1 ratio is mixed, and curing 30min after cast film forming, under the conditions of 90 DEG C can prepare.The reality of the material of the mantle of the present invention
Example includes DOW CORNING sylgard 184.
In the apparatus of the present, the groove 3 of grooved drum plate 1 includes the Uncrossed passage of N (N >=1) item, and groove width is
100-5000 μm, depth of rebate is 100-5000 μm, and the length of groove can be designed according to actual requirement；Further preferably
Ground, groove width are 200-1500 μm, and gash depth is 200-1500 μm；It is highly preferred that gash depth is 400-600 μm, with
Conventional method is compared, and is conducive to reduce amount of samples, such as amount of samples can be conducive to detect down to 20 μ L.
In the apparatus of the present, 3 surface of groove can not be modified, can also be according to the difference of capture molecular property
It is modified so that capture is easier to be attached to its surface with molecule.
In the apparatus of the present, at least two test points are equipped in every passage of the groove 3, have been loaded respectively
Different capture molecules.Spacing between test point is 0.3-10mm, is preferably 1-5mm, more preferably 1-2mm.So may be used
To ensure testing efficiency, and ensure that each test point will not influence each other and reduce detection sensitivity.
In the present invention, the groove 3 of grooved drum plate 1 be by being machined, being molded, photoengraving, Soft lithograph are prepared.Ditch
Frid 1 and bottom plate 2 can be combined the two by thermal bonding, ultrasonic bond or the methods of being mechanically fixed, but are not limited solely to
Method is stated, any one can be by method that the two is glued together all within the protection of the present invention.
In the present invention, the test point load of groove 3 has different capture molecules, will capture to use by point sample mode and divide
Son is supported in groove 3.The capture molecule of the present invention includes but is not limited to antigen, antibody, and/or probe；Capture, which is used, to divide
Load combination of the son at test point includes but is not limited to chemical bond, intermolecular interaction etc.；Point sample mode can
It is simple to operate using point sample instrument or manual point sample.For example, from left to right existed successively using Biodot XYZ3060 point sample instruments
Point sample in the groove of grooved drum plate, specific point sample parameter are that a spacing can be 0.3-10mm, are preferably 1-5mm, more preferably 1-
2mm；Drop amount can be 10-1000nl, be preferably 30-800nl, more preferably 50-100nl；Needle point apart from grooved drum plate 3 away from
It is placed a period of time within 2mm, point sample time control room temperature after 0.6-0.9ms, point sample is open, such as 1-2h or so,
So as to improve bond quality.Further, since batch production, therefore environment between production can be controlled, make detection device suitable for item
Sealing is combined or encapsulated under part.
In use, the mode being detected includes but is not limited to CCD and scanner, signal producing method includes but not office
It is limited to chemiluminescence, fluorescence and colour developing.
In the present invention, according to the species and property of serum to be checked, detection target, reagent and label and to colour developing
As a result needs can select following different mode：
1st, sandwich method：According to requirement of experiment using double antibody or dual-antigen sandwich method, the known antigens in this device are utilized
Or antibody, it adds in unlabelled sample and interacts (containing antibody or antigen), be eventually adding the antigen or antibody of mark,
It is developed the color or the detection that shines, has at signal for the positive, be feminine gender at no signal, can be quantified according to colour developing or luminous intensity
Research.
2nd, indirect method：Known reagent, such as antigen in this device add in unlabelled sample (such as antibody) and carry out mutually
Effect, the secondary antibody for being eventually adding mark are developed the color or chemiluminescence detection, are had at signal for the positive, are feminine gender at no signal,
Can quantitative study be carried out according to colour developing or luminous intensity.
3rd, competition law：Known antigen or antibody in this device add in the antigen or antibody of known mark in the sample,
Mixed with unlabelled antigen or antibody, it is known that antigen or antibody competition it is mutual with the antibody in groove or antigen
Effect adds in substrate, and signal strength is feminine gender, is the positive without signal or the weak place of signal, can according to colour developing or luminous intensity into
Row quantitative study.
Below with reference to embodiment, the present invention will be described in detail.
It is prepared by embodiment 1- chips
Polystyrene board is taken, injection moulding is used to portray 1 width at a surface thereof as 1000 μm of S-shaped groove, wherein ditch
Groove depth is 200 μm, and prepared by such grooved drum plate completes, and notch is formed in the correspondence position of another side；Prepare AFP (first tire eggs
In vain) antibody, CEA (carcinomebryonic antigen) antibody, (wherein AFP resists the sampling liquid (abbreviation sample liquid) of CA125 (glycogen chain antigen 125) antibody
Body, CEA antibody, the concentration of CA125 antibody are 20 μ g/ml), from left to right existed successively using Biodot XYZ3060 point sample instruments
Point sample in the groove of grooved drum plate, specific point sample parameter are that a spacing is 2mm, and drop amount is 50nl, needle point apart from point template away from
1-2h or so is placed within 2mm, point sample time control room temperature after 0.6-0.9ms, point sample is open；It separately takes same with grooved drum plate
The polystyrene board of sample ruler cun, by grooved drum plate and bottom plate using Mechanical Method sealing-in, obtains micro-fluidic chip detection dress as bottom plate
It puts.
It is prepared by embodiment 2- chips
Dimethyl silicone polymer plate is taken, mechanical processing method is used to portray 1 width at a surface thereof as 100 μm of S shape ditches
Slot, wherein gash depth are 400 μm, form notch in the correspondence position of another side, prepared by such grooved drum plate completes；Second is configured
Liver surface antibody, hepatitis A surface antibody, sampling liquid (wherein hepatitis B surface antibody, hepatitis A surface antibody, third of hepatitis surface antibody
The concentration of liver surface antibody is 30 μ g/ml), with Biodot XYZ3060 point sample instruments from left to right successively in the groove of grooved drum plate
Interior point sample, specific point sample parameter are that a spacing is 2mm, and drop amount is 50nl, distance of the needle point apart from point template within 2mm,
Point sample time control room temperature after 0.6-0.9ms, point sample is open to place 1-2h or so；Separately take poly- two with grooved drum plate same size
Methylsiloxane, by grooved drum plate and bottom plate using ultrasonic bonding sealing-in, obtains flexible biological probe as bottom plate.
It is prepared by embodiment 3- chips
Poly (methyl methacrylate) plate is taken, Soft lithograph method is used to portray 1 width at a surface thereof as 5000 μm of curved shape grooves, wherein ditch
Groove depth is 800 μm, forms notch in the correspondence position of another side, prepared by such grooved drum plate completes；AFP antibody, CEA are configured
Antibody, PSA (prostate-specific antigen) antibody, CA125 carbohydrate antigens antibody, CA19-9 sugar antigens antibody and CA15-3 sugar chains
(wherein AFP antibody, CEA antibody, PSA antibody, CA125 carbohydrate antigens antibody, CA19-9 sugar antigens resist the sampling liquid of antigen-antibody
The concentration of body and CA15-3 carbohydrate antigen antibody is 30 μ g/ml), pipetted with pipettor the sampling liquid configured from left to right according to
The secondary point sample in the groove of grooved drum plate, specific point sample parameter are that a spacing is 2mm, and drop amount is 500nl, contact point sample, point
Room temperature is open after sample places 1-2h or so；It is another to take with the poly (methyl methacrylate) plate of grooved drum plate same size as bottom plate, by grooved drum plate and
Bottom plate uses mechanical Fixing Method sealing-in, obtains flexible biological probe.
It is prepared by embodiment 4- chips
Polyvinyl chloride panel is taken, photoetch method is used to portray 1 width at a surface thereof as 1000 μm of S-shaped grooves, wherein ditch
Groove depth is 1000 μm, forms notch in the correspondence position of another side, prepared by such grooved drum plate completes；Concentration has been configured as 20 μ
(wherein hav antigen, hepatitis B antigen and hepatitis C antigen is dense for the sampling liquid of the hav antigen of g/ml, hepatitis B antigen and hepatitis C antigen
Degree is 20 μ g/ml), pipetting sampling liquid with pipettor, from left to right the point sample in the groove of grooved drum plate, specific point sample are joined successively
Number is that a point spacing is 2mm, and drop amount is 500nl, and contact point sample, room temperature is open after point sample places 1-2h or so；It is another to take and ditch
The polyvinyl chloride panel of frid same size, by grooved drum plate and bottom plate using ultrasonic bonding sealing-in, obtains miniflow as bottom plate
Control chip-detecting apparatus.
Embodiment 5- chip applications
The AFP that concentration is 75ng/ml is added in into groove by the notch of the detection device prepared in embodiment 1,
The standard items of CEA, CA125 react at room temperature 30min, absorb remaining standard items, add in enzyme labelled antibody, react at room temperature 30min, clearly
It washes 3 times, drains pipeline, add in luminescent solution (Millipore Western Blot chemiluminescent HRP substrate ECL luminescent solutions, goods
Number：WSKLS0050, similarly hereinafter), chip is put into chip reading instrument (instrument producer：Beijing Yuanpinghao Biological Technology Co., Ltd.,
Model：Tanon 5200) it is detected.Test result is shown in Fig. 4, we can see that positive (hair is presented in each index from figure
Light part is the positive), which can measure AFP, CEA, CA125 simultaneously, compared with the ELISA method of individual event, substantially reduce
Detection time reduces testing cost, and can realize automation, simplifies operating procedure.
Embodiment 6- chip applications
Concentration is added in into groove by the notch of the detection device prepared in embodiment 2 is (from left to right) successively
The hepatitis B of 9.375ng/ml, 18.75ng/ml, 37.5ng/ml, hepatitis A, the standard items of hepatitis react at room temperature 30min, absorb surplus
Remaining standard items add in enzyme labelled antibody, react at room temperature 30min, clean 3 times, drain pipeline, add in luminescent solution, chip is put into core
Piece reading apparatus is detected.Test result is shown in Fig. 5, we can see that the positive is presented in each index, (luminous component is i.e. from figure
For the positive), with the increase of standard concentration, luminous intensity enhancing, which can measure hepatitis B, hepatitis A and hepatitis simultaneously.
Embodiment 7- chip applications
By the notch of the detection device prepared in embodiment 3 added in into groove AFP that concentration is 18.75ng/ml,
CEA, PSA (prostate-specific antigen), CA125 carbohydrate antigens, the mixing mark of CA19-9 sugar antigens and CA15-3 carbohydrate antigens
Quasi- product react at room temperature 30min, absorb remaining standard items, add in enzyme labelled antibody, react at room temperature 30min, clean 3 times, drain pipe
Road adds in luminescent solution, chip is put into chip reading instrument and is detected.Test result is shown in Fig. 6, from figure we can see that each
Index is presented positive (luminous component is the positive), the device can measure simultaneously AFP, CEA, PSA, CA125, CA19-9 and
The associated antibodies of CA15-3.
Embodiment 8- chip applications
By the notch of the detection device prepared in embodiment 5 added in into groove hepatitis A that concentration is 37.5ng/ml,
Hepatitis B and c-hepatitis antibody standard items react at room temperature 30min, absorb remaining standard items, add in enzyme labelled antibody, react at room temperature 30min,
Cleaning 3 times drains pipeline, adds in luminescent solution, chip is put into chip reading instrument and is detected.Test result is shown in Fig. 7, from figure
We can see that positive (luminous component is the positive) is presented in each index, which can measure hepatitis A, hepatitis B and third simultaneously
The associated antibodies of liver.
Claims (14)
1. a kind of microflow controlled biochip detection device, which is characterized in that the detection device includes grooved drum plate, bottom plate；It is described
Grooved drum plate includes first surface and the second surface opposite with first surface, and close by the first surface and the bottom plate
Sealed knot closes；The first surface of the grooved drum plate be equipped at least one Uncrossed groove, every groove with the grooved drum plate
First surface border between leave distance, and at least one test point is equipped in every groove, on each test point point
Fu Zai there be a kind of capture molecule；The second surface of the grooved drum plate is equipped with notch, and the notch is arranged at described second
With the corresponding position in every groove both ends on surface, and connected with the groove, it is to be tested for being added into every groove
Sample；
Realize that sealing combines by thermal bonding, ultrasonic bond or mechanical means between the first surface and the bottom plate.
2. detection device according to claim 1, which is characterized in that at least two test points, institute are equipped in every groove
Load has different capture molecules respectively at least two test points stated.
3. detection device according to claim 2, which is characterized in that the capture is selected from antigen, antibody or spy with molecule
Pin.
4. according to the detection device described in claim 1 or 2 or 3, which is characterized in that the test point is used by that can adsorb capture
The material of molecule is made.
5. detection device according to claim 4, which is characterized in that the capture that can adsorb is with the material of molecule
Simple glass, polystyrene, polyvinyl chloride, makrolon, organic glass, dimethyl silicone polymer or they through active group
The modified material of group.
6. detection device according to claim 1, which is characterized in that the width of the groove is 100-5000 μm, described
The depth of groove is 100-5000 μm；The test point is located at the bottom of the groove.
7. the detection device according to claim 1 or 6, which is characterized in that the test point is by directly forming the ditch
The location point or the position formed by other objects being attached in the trench bottom surfaces that the region of groove bottom is formed
Point.
8. detection device according to claim 1, which is characterized in that the first surface is equipped with a Uncrossed ditch
Slot or the first surface are equipped at least two Uncrossed grooves.
9. detection device according to claim 8, which is characterized in that the groove is S-type or snakelike.
10. detection device according to claim 1, which is characterized in that the grooved drum plate and bottom plate are all tablet, Huo Zhewei
Plate with the radian that matches.
11. detection device according to claim 1, which is characterized in that the notch is tapered, conical lower portion with it is described
Groove connects.
12. detection device according to claim 11, which is characterized in that sealant is additionally provided on the second surface, is used
In the closing notch.
13. the preparation method of the detection device according to any one of claim 1-12, which is characterized in that including walking as follows
Suddenly：
A, at least one groove is formed on the first surface of grooved drum plate, and on the second surface of the grooved drum plate and per bar ditch
The corresponding position in slot both ends opens up notch, and the notch is connected with the groove, in order to add sample to be tested into groove
Product；
At least one capture is put to precalculated position in the trench respectively with molecule by point sample mode b, it is at least one to be formed
Load has a kind of test point of capture molecule；
C, the first surface of the bottom plate and the grooved drum plate is sealed and combined, obtain the detection device.
14. a kind of method that detection device using any one of claim 1-12 carries out biological detection, including：
The first step：Detected sample is injected into the groove by the notch of described groove one end；
Second step：The detected sample is examined with the capture being supported on the test point with the interaction of molecule
It surveys.
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| CN201510294370.5A CN104897654B (en) | 2015-06-02 | 2015-06-02 | A kind of microflow controlled biochip detection device and preparation method |
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| CN201510294370.5A CN104897654B (en) | 2015-06-02 | 2015-06-02 | A kind of microflow controlled biochip detection device and preparation method |
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