CN102239005A

CN102239005A - Integrated detection chip and application method thereof

Info

Publication number: CN102239005A
Application number: CN2009801443803A
Authority: CN
Inventors: 王战会; 陈坦
Original assignee: TIANJIN MNCHIP TECHNOLOGIES CO LTD
Current assignee: TIANJIN MNCHIP TECHNOLOGIES CO LTD
Priority date: 2009-11-03
Filing date: 2009-11-03
Publication date: 2011-11-09
Also published as: WO2011054140A1

Abstract

An integrated detection chip and an application method thereof are disclosed. Said chip comprises an upper layer (1) and a lower layer (2) which are connected each other. The lower layer of said chip has a group of chambers (21,22,23,24,25,26,27), micro-fluidic circuits (28,29) connecting each chamber, micro-reflective surfaces (210) and vent ports (211). Said chip can automatically perform pre-processing, quantificational transmission, reaction and detection of the sample solutions. Said chip is applied to the analysis and detection field, for example, biological analysis, medical detection, environmental pollution monitoring and food & drug detection.

Description

Integrated detection chip and application method thereof

Specification

Title of Invention:Detect integrated chip and application process technical field

Technical field

[1] the invention belongs to analyze detection field, and in particular to a kind of pre-treatment of collection sample, the quantitative conveying of sample, react and be detected on integral multi-functional multiple determination integrated chip and application process.

Background technology

[2] numerous challenges of the 21 century in fields such as bio-medical analysis, medical diagnosis on disease, environmental monitoring, food and drug safeties are faced with, spot sampling analysis, quick detection and patient such as are tested oneself at the proposition of demand, and inspection and quarantine analysis means and equipment are proposed with higher requirement.These new demands constantly proposed are met, development miniaturization, integrated and portability inspection and quarantine instrument and equipment is just had to.

[3] origin for the automated analysis equipment that inspection and quarantine at present is used can trace back to last century the fifties, have passed through semicentennial development and have been achieved with integrated and automation.Such as automatic biochemistry analyzer is to calculate the sampling in biochemical analysis, reagent adding, mixing, insulation, colorimetric, result with reporting that the part or all of of these steps is completed by imitating manual manipulator.But existing automatic biochemistry analyzer is bulky, expensive, complex operation, also needs to be equipped with professional equipment and carries out the pre-treatment of sample, so needing the central laboratory installed in large hospital, is operated by trained professional.In addition, in order to improve detection efficiency and reduction testing cost, generally requiring to gather up a fairly large number of a collection of sample to carry out united analysis detection, so detection cycle is longer.The characteristics of large automatic Biochemical Analyzer used in Hospitals at Present, is difficult to meet spot sampling analysis, quick detection and patient and the demand such as tests oneself.

To the disclosure of invention

Technical problem

[4] integrated chip is detected object of the present invention is to provide one kind, it can overcome the defect of existing detection device, can automatically complete whole detection process, and detection cycle is short, one-time detection just can obtain multiple Testing index

[5] it is a further object of the present invention to provide a kind of application process, it can be detected using above-mentioned integrated chip. Technical solution

[6] a kind of detection integrated chip, the chip be divided into above and below two layers, described chip upper strata is connected with described chip lower floor；The chip upper strata is used for the through hole of sample pipetting volume provided with one group；Described chip lower floor is provided with fluid channel, micro- reflecting surface and the passage between one group of groove and each groove of connection.

[7] a kind of application process using above-mentioned detection integrated chip, it comprises the following steps：

[8] sample solution via through holes are injected into described sample cell by (1)；

[9] (2) start the described chip of motor rotation；

[10] (3) sample solution under the action of the centrifugal force, realizes separation of solid and liquid；

[11] liquid in (4) sample flows into reactive tank, is reacted with described reaction reagent；

[12] (5) carry out in situ detection by the detection device supporting with chip in reactive tank.

Beneficial effect

[13] using after such scheme, the detection integrated chip collection sample pre-treatments of the present invention, sample delivery, react and be detected on one, sample pre-treatments equipment and manipulator without being equipped with specialty, it is simple to operate, whole detection process full-automation is completed, without professional's operation, it is possible to achieve patient tests oneself.A piece of chip, one-time detection is obtained with multiple Testing index, and detection cycle is short.Multi-functional multiple determination integrated chip volume is small, integrated with miniaturization detection device, can develop miniaturization, integrated and portability detection device, realizes that spot sampling is analyzed.

[14] in addition, the multi-functional multiple determination integrated chip of the present invention can be used for field of biological medicine, and full-automatic quick detection is carried out to body fluid such as the whole blood of human or animal, blood plasma, urine, saliva, seminal fluid, spinal cord, amniotic fluid.The multi-functional multiple determination integrated chip of the present invention can be used for environment measuring field, and the organic or inorganic pollutant in environment is used for quickly detecting.The multi-functional multiple determination integrated chip of the present invention can be used for field of food safety, and the poisonous and harmful substance in food, bacterium, virus etc. are used for quickly detecting.The multi-functional multiple determination integrated chip of the present invention can be used for pharmacy, chemical field, and various drug ingredients, chemical products are used for quickly detecting.

Brief description of the drawings

[15] Fig. 1 is the top view on chip upper strata；

[16] Fig. 2 is the schematic top plan view of chip lower floor；

[17] Fig. 3 after to be sample add with contrast tested liquid chip start the moment schematic diagram that rotates with motor； [18] Fig. 4 is that sample flows through spiral fluid channel and reservoir proceeds by the schematic diagram of separation of solid and liquid；

[19] Fig. 5 is sample after separation of solid and liquid by going in ring runner and divergence expression runner enters the schematic diagram of reactive tank；

[20] Fig. 6 is the schematic diagram that isothermal reaction and optical detection are carried out after reactive tank is filled.

Embodiments of the present invention

[21] below so that whole blood biochemistry is detected as an example, and it is described with reference to the accompanying drawings the application method of multi-functional multiple determination integrated chip of the invention.

[22] if thing concentration to be detected is higher in using sample to be detected, pre-dilution to suitable concentration, it can be then added in chip and detect outside chip.Such as the Analysis of Biochemical of blood, pre-dilution can be carried out to blood and is then added to detection in chip.In addition, this chip can be used for field of biological medicine, many indexes in the body fluid such as the whole blood of human or animal, blood plasma, urine, saliva, seminal fluid, spinal cord, amniotic fluid are carried out with full-automatic quick detection.Furthermore, this chip can be also used for environment measuring field, and the organic or inorganic pollutant in environment is used for quickly detecting.This chip can be also used for field of food safety, and the poisonous and harmful substance in food, bacterium, virus etc. are used for quickly detecting.This chip can be used for pharmacy, chemical field, and various drug ingredients, chemical products are used for quickly detecting.

[23] in order to make the purpose , technical scheme and advantage of the present invention be clearer, below in conjunction with drawings and Examples, the present invention will be described in further detail.It should be understood that, although depicting the present invention by embodiment, it will be appreciated by the skilled addressee that the present invention has many deformations and the spirit changed without departing from the present invention, and the specific embodiments described herein are merely illustrative of the present invention, it is not intended to limit the scope of the present invention

[24] embodiment one：Blood biochemical is analyzed

[25] as shown in Figure 1, 2, the present invention discloses a kind of detection integrated chip, and the chip is made using transparent plastic as raw material by way of die casting.This chip includes upper strata 1 and lower floor 2, wherein：

[26] upper strata 1 is provided with two through hole 11,12, and through hole 11 is used to add sample, and through hole 12 is used to add contrasting fluid.

[27] lower floor 2 is provided with sample cell 21, reservoir 22, contrast reference liquid bath 23, one group of reaction detection groove 24 and one group of contrast tested liquid reaction detection groove 25, one group of auxiliary tank 26 for self-inspection and detection correction, fluid channel 29, micro- reflecting surface 210 and steam vent 211 between one group of overflow launder 27, the spiral fluid channel 28 for separation of solid and liquid, each groove of connection.

[28] sample cell 21 is provided with steam vent 211, and sample cell 21 is located at chip and leans on paracentral position, and sample passes through core The through hole 11 on piece upper strata 1 is injected into sample cell 21.

[29] spiral fluid channel 28, entrance is connected with sample cell 21, and outlet is connected with reservoir 22.The width of spiral fluid channel 28 gradually broadens from the inlet to the outlet, and the fluid channel width ratio in entrance and exit is 1:1.The depth of spiral fluid channel 28 is gradually deepened, and the fluid channel depth scale in entrance and exit is 1:1.The entrance of spiral fluid channel 28 is small, plays a part of limiting sample flow, is separated between the solid insoluble in sample is had enough inch under centrifugal force before outflow reservoir 22 with liquid.Spiral fluid channel 28 is connected with reservoir 22 extends the distance that sample solution flows through, and is conducive to being completely separated with liquid before outflow reservoir 22 the solid insoluble that spiral fluid channel 28 do not separate also with liquid is flowed through.

[30] reservoir 22 in a ring, radial distribution.

[31] reservoir 22 is connected by arc fluid channel 292 with reactive tank 24.

[32] a diameter of 0.1mm of reactive tank 24, depth is lmm.Reactive tank 24 is located remotely from the circumference in the center of circle, and shape can be circular, square, rectangle or polygon, and in the present embodiment, it is circle.

[33] contrast is provided with steam vent 211 with reference to liquid bath 23, and tested liquid is entered in the groove by the import 12 on 1.With reference to liquid bath 23 with contrast tested liquid reaction detection groove 25 be to be connected by runner 291.

[34] various freeze-dried reagents are preinstalled with reaction detection groove 24, in the present embodiment, the freeze-dried reagent pre-installed in the reaction detection groove be for detection blood parameters include ALT (ALT), aspartate amino transferase (AST), γ-paddy ammonia phthalidyl transferase (γ-Ο Τ), alkaline phosphatase (ALP), total bilirubin (TBI L), bilirubin direct（DBIt), total protein (TP), albumin (Alb), urea (Urea), creatinine (Cr), uric acid (UA), glucose (Glu), T-CHOL (TC), triglycerides（TG) 、

HDL（HDL), the low albumen of low-density（VLDL), VLDL（LDL), serum magnesium（Mg), serum potassium（K), serum sodium（Na), serum chloride（C1), serum calcium（Ca), serum paraoxonase（P), serum levels of iron（Fe), serum ammonia（NH), carbon dioxide（C02) .

[35] auxiliary tank 26 is used for the self-inspection and correction of the detection device supporting with chip, and this group of auxiliary tank 26 is located remotely from the circumference in the center of circle.The distance in the center of auxiliary tank 26 to the center of circle is identical with distance of the reaction detection groove 24 away from the center of circle.A diameter of 2mm of auxiliary tank 26.

[36] overflow launder 27 includes contrast tested liquid overflow launder 271 and sample overflow launder 272.It is connected with reference to liquid bath 23 with contrast tested liquid overflow launder 271 by runner 291.

[37] sample overflow launder 272 is connected by annular channel 293, arc fluid channel 292 with reservoir 22. [38] micro- reflecting surface 210, auxiliary tank 26 are pinpoint for reaction detection groove optics, and micro- reflecting surface 210 is located at around reaction detection groove 24.The width of micro- reflecting surface 210 is 0.1mm.The central shaft angle of micro- reflecting surface 210 and reaction detection groove 24 is 0 degree.

[39] aperture 211 is steam vent.Its diameter is between 0.1mm to 2mm.

[40] use inch, first bled liquid from the access of the finger tip of human or animal, vein or other positions, then by 1:100 dilution proportion, and inject sample cell 21 from through hole 11.Contrast tested liquid is injected contrast with reference to liquid bath 23 from through hole 12;

[41] coordinate shown in Fig. 3, chip is fixed on motor, is placed in 37 degrees Celsius of constant temperature chambers, chip rotation, 4000 rpms of speed are driven by motor；The particulate materials such as the haemocyte that blood sample enters in spiral fluid channel 28, blood from sample cell 21 under the influence of centrifugal force start to separate with liquid, and contrast tested liquid enters air stream enter runner 291 from contrast with reference to liquid bath 23 under the influence of centrifugal force；

[42] coordinate shown in Fig. 4, chip continues to rotate under motor drive, and blood sample can be stepped into reservoir 22 from sample cell 21 by spiral fluid channel 28.With this contrast tested liquid detect tank 25 and contrast tested liquid overflow launder 271 are stepped into inch contrast tested liquid by runner 291.

[43] coordinate shown in Fig. 5, contrast tested liquid detect tank 25 and contrast tested liquid overflow launder 271 are all filled with.Blood sample under the influence of centrifugal force, can little by little separate haematoblast and blood plasma two layers, haemocyte is trapped on the side wall in the remote center of circle of spiral fluid channel 28 and reservoir 22 in spiral fluid channel 28 and reservoir 22.When amount of liquid storage gradually increases in reservoir 22, reach certain volume inch, blood plasma can enter annular channel 293 by arced flow path 292 under the action of the centrifugal force, reaction detection groove 24 is full of one by one by the divergence expression runner 294 being connected with each reaction detection groove 24 again, the reagent pre-installed in dissolving reaction detection groove 24, and start reaction

[44] coordinate shown in Fig. 6, all without liquid in sample cell 21, spiral fluid channel 28, contrast reference liquid bath 23 and runner 291.All reaction detection grooves 24 have been filled with fluid sample, and sample overflow launder 272 is also filled.Chip continues to keep rotation, and each reactive tank 24 is accurately positioned by the micro- reflecting surface 210 adjacent with reaction detection groove 24, and the reaction in each reaction detection groove 24 is detected by spectrophotometer, provides testing result.This inch, reaction detection groove 24 is also referred to as cell.

[45] embodiment two：Compared with embodiment one, difference is：The width of the porch of spiral fluid channel 28 and the width ratio in exit are to 1:10, the depth of the porch of spiral fluid channel 28 and the depth scale in exit For 1: 10.The a diameter of 10mm depth of reactive tank 24 is 10mm.A diameter of 10mm of auxiliary tank 26.The width of micro- reflecting surface 210 is to 5mm.The central shaft angle of micro- reflecting surface 210 and reactive tank 24 is 90 degree.

[46] embodiment three：Compared with embodiment one, difference is：The width of the porch of spiral fluid channel 28 and the width ratio in exit are 1:5.The depth of the porch of spiral fluid channel 28 and the depth scale in exit are 1: 2.The a diameter of 2mm depth of reactive tank 24 is 5mm.The width of micro- reflecting surface 210 is lmm.The central shaft angle of micro- reflecting surface 21 0 and reactive tank 24 is 45 degree.

[47] example IV：Water quality detection

[48] using transparent plastic as raw material, chip is made by way of die casting, and is pre-installed in reactive tank 24 for detecting Organic substance in water（Such as phenol）, heavy metal ion（Such as lead, copper, iron）, residues of pesticides（Such as organophosphor）Reagent.1 milliliter of water sample is taken to be added in sample cell 21.The injection contrast of contrast tested liquid with reference to liquid bath 23.Chip is fixed on motor, starts motor and starts rotation.The steps such as sample delivery, separation, reaction detection are similar with embodiment one.

[49] embodiment five：Food hygiene detection

[50] using transparent plastic as raw material, chip is made by way of die casting, and is pre-installed in reactive tank 24 for detecting microorganism in food（Such as Escherichia coli）, additive（Such as saccharidase preparation）, residues of pesticides（Such as organophosphor, carbamate pesticide）, pollutant（Such as inorganic arsenic, formaldehyde, cyanide, nitrite）, protein etc. reagent.It can be added to for liquid sample with direct sample in chip.Solid sample needs to crush, then adds diluted dissolving, is then added in chip.It is special to use after various abstraction techniques extract test substance from food, it is then added to chip detection.Detection method be the same as Example one.

[51] the foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, any modifications, equivalent substitutions and improvements made within the spirit and principles of the invention etc., should be included in the scope of the protection

Claims

Claims

[Claim 1] one kind detection integrated chip, it is characterised in that：Described chip be divided into above and below two layers, described chip upper strata is connected with described chip lower floor；The chip upper strata is used for the through hole of sample pipetting volume provided with one group；Described chip lower floor is provided with fluid channel, micro- reflecting surface and the steam vent between one group of groove and each groove of connection.

[Claim 2] detects integrated chip as claimed in claim 1, it is characterised in that：The groove of described chip lower floor is to be connected between sample cell, reservoir, contrast reference liquid bath, one group of reaction and detect tank, one group of auxiliary tank, one group of overflow launder for self-inspection and detection correction, above-mentioned each groove by fluid channel.

[Claim 3] detects integrated chip as claimed in claim 1, it is characterised in that：Described chip lower floor further comprises the spiral fluid channel for separation of solid and liquid.

[Claim 4] surveys integrated chip as claimed in claim 2 or claim 3, it is characterised in that：The sample cell of described chip lower floor, positioned at by paracentral position, described sample is injected into by the through hole on described chip upper strata in described sample cell.

[Claim 5] detects integrated chip as claimed in claim 3, it is characterised in that：The spiral fluid channel of described chip, entrance is connected with described sample cell, and outlet is connected with described reservoir

[Claim 6] detects integrated chip as claimed in claim 3, it is characterised in that：The width of described spiral fluid channel gradually broadens from the inlet to the outlet, and the fluid channel width ratio in entrance and exit is 1:1 to 1: 10.

[Claim 7] detects integrated chip as claimed in claim 6, it is characterised in that：The width of the porch of described spiral fluid channel and the width ratio in exit are 1:5.

[Claim S] detects integrated chip as claimed in claim 3, it is characterised in that：Described spiral fluid channel depth is gradually deepened, and the fluid channel depth scale in entrance and exit is 1:1 to 1: 10.

[Claim 9] detects integrated chip as claimed in claim 8, it is characterised in that：The depth of the porch of described spiral fluid channel and the depth scale in exit are 1: 2.

[Claim 10] detects integrated chip as claimed in claim 3, it is characterised in that：The reservoir of described chip lower floor is connected with the outlet of described spiral fluid channel, in a ring, radial distribution. Detection integrated chip as claimed in claim 2, it is characterised in that：The reactive tank diameter of described chip lower floor is between 0.1mm to 10mm.

Detection integrated chip as described in claim 2 or 11, it is characterised in that：The depth of the reactive tank of described chip lower floor is between lmm to 10mm.

Detection integrated chip as claimed in claim 2, it is characterised in that：Reaction reagent is preinstalled with described reactive tank.

Detection integrated chip as claimed in claim 13, it is characterised in that：Described reaction reagent is reaction reagent, the dry powder-shaped reaction reagent of liquid, or the reaction reagent of prepackage liquid, then add one or more kinds of lyophilized reaction reagents after described reactive tank situ is lyophilized or reaction reagent original position of pre-mounted part liquid is lyophilized.

Detection integrated chip as claimed in claim 2, it is characterised in that：The contrast of described chip lower floor is connected with reference to liquid bath by fluid channel with one group of reactive tank, and described contrast tested liquid is used for the contrast reference of testing result.

Detection integrated chip as claimed in claim 2, it is characterised in that：The auxiliary tank of described chip lower floor is used for the self-inspection and correction of the detection device supporting with the chip, and this group of auxiliary tank is located remotely from the circumference in the center of circle.

Detection integrated chip as claimed in claim 16, it is characterised in that：The distance in center to the center of circle of one group of described auxiliary tank is identical with distance of the described reactive tank away from the center of circle.Detection integrated chip as described in claim 16 or 17, it is characterised in that：The diameter of described auxiliary tank is between lmm to 10mm.

Detection integrated chip as claimed in claim 2, it is characterised in that：Micro- reflecting surface of described chip is to be used to react pinpoint with detect tank optics, and it is located at around described reactive tank.

Detection integrated chip as described in claim 1 or 19, it is characterised in that；Described micro- reflecting surface width is between 0. lmm to 5mm.

Detection integrated chip as claimed in claim 19, it is characterised in that：Described micro- reflecting surface is with described reactive tank central shaft angle between 0 to 90 degree.

Detection integrated chip as claimed in claim 1, it is characterised in that：Described chip upper strata Through hole be two.

Detection integrated chip as claimed in claim 1, it is characterised in that：The raw material of described chip are plastics, silica gel, metal, alloy, glass or silicon.

Detection integrated chip as claimed in claim 1, it is characterised in that：Using Nian Jie or with being connected by the way of ultrasonic bonding between described chip upper strata and lower floor.

With the application process of any described detection integrated chip of claim 1 ~ 24, it comprises the following steps：

(1) sample solution via through holes are injected into described sample cell；

(2) the described chip of motor rotation is started；

(3) sample solution under the action of the centrifugal force, realizes separation of solid and liquid；

(4) liquid in sample flows into reactive tank, is reacted with described reaction reagent；

(5) in situ detection is carried out in reactive tank by the detection device supporting with chip.Application process as claimed in claim 25, it is characterised in that：In step（2) in, the rotating speed of described motor is between 1000 to 10000 rpms.

Application process as claimed in claim 25, it is characterised in that：In step（3) in, solid insoluble is separated with described reservoir process with liquid flowing through described spiral fluid channel in sample, be trapped in described in spiral fluid channel and described reservoir in.

Application process as claimed in claim 25, it is characterised in that：In step（4) in, liquid flows through spiral fluid channel, reservoir, the fluid channel being connected with reactive tank successively in sample, eventually enters into reactive tank.

Application process as claimed in claim 25, it is characterised in that：In step（3) 、（4

) (5) same inch for carrying out, described chip is in rotation status all the time.

Application process as claimed in claim 25, it is characterised in that：Disposable full-automation completes pre-treatment, quantitative conveying, reaction and the detection process of sample solution in the chips.Application process as claimed in claim 25, it is characterised in that：In step（5) in, supporting equipment is spectrophotometer, fluorescence photometer, photoelectric tube with chip.

Application process as described in claim 25 or 31, it is characterised in that：In step（5) in

, supporting detection method is AAS, fluorescence method, chemoluminescence method with chip.