CN102998439A - Micro-fluidic paper chip for simultaneously detecting glucose, uric acid, triglycerides and cholesterols, and its manufacturing method - Google Patents
Micro-fluidic paper chip for simultaneously detecting glucose, uric acid, triglycerides and cholesterols, and its manufacturing method Download PDFInfo
- Publication number
- CN102998439A CN102998439A CN2011102702132A CN201110270213A CN102998439A CN 102998439 A CN102998439 A CN 102998439A CN 2011102702132 A CN2011102702132 A CN 2011102702132A CN 201110270213 A CN201110270213 A CN 201110270213A CN 102998439 A CN102998439 A CN 102998439A
- Authority
- CN
- China
- Prior art keywords
- micro
- cholesterol
- oxidase
- refill sheet
- glucose
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention provides a micro-fluidic paper chip for simultaneously detecting glucose, uric acid, triglycerides and cholesterols, and its manufacturing method. The paper chip is a novel personalized diagnosis device, can rapidly, cheaply and simply detect multiple components without instrument equipment or professional people, can be operated by a patients or their families, and can momentarily and rapidly detect the concentrations of the glucose, the uric acid, the triglycerides and the cholesterols in blood.
Description
Technical field
The present invention designs a kind of three-dimensional microflow control refill sheet, is used for detecting simultaneously glucose, uric acid, triglyceride and cholesterol.
Background technology
Blood parameters can reflect the health condition of human body, also is the important evidence of diagnosis and treatment.Along with the raising of people's living standard, the incidence of disease of the diseases such as diabetes, high fat of blood and gout significantly rises the serious harm human health.Therefore, the detection of glucose in blood, triglyceride, cholesterol and uric acid has become test item conventional in the hospital.Large-scale Biochemical Analyzer usually can be finished simultaneously the haemoconcentration of above-mentioned substance and measure, but relatively wastes time and energy, and cost is higher, and relies on the operating personnel of specialty.Because diabetes, high fat of blood and gout all belong to chronic disease, need long-term more frequently check, so instrumental method has been brought inconvenience to the patient, be unfavorable for that also find and early diagnosis the morning of conditions of patients.Although, at present for glucose in blood, triglyceride, cholesterol and uric acid, developed fast and convenient detection method, there is not a kind of method to detect above-mentioned four kinds of materials in high flux ground simultaneously.
Summary of the invention
Actual conditions for present glucose, triglyceride, cholesterol and uric acid detection, adapt to the development trend that family is detected, the present invention combines the enzyme process detection system with microflow control technique, set up a kind of high flux three-dimensional microflow control refill sheet that can detect simultaneously glucose, triglyceride, cholesterol and uric acid.This chip need not instrument and equipment and professional operating personnel, and is highly sensitive, and specificity is good, and is simple to operate, and visual phenomenon is a kind of efficient urine uric acid detection method that patient family and various medical institutions use that is fit to very much.
The present invention is by the following technical solutions:
A kind of micro-fluidic refill sheet that detects simultaneously glucose, uric acid, triglyceride and cholesterol, the refill sheet is by the three-dimensional micropore road system that photoetching filter paper and punching double faced adhesive tape stack form, include the specific oxidation enzyme,, peroxidase, 4-AA and N-ethyl-N-(3-sulfoacetic acid)-3 aminoanisoles.
The specific oxidation enzyme is glucose oxidase, urate oxidase, GPO and cholesterol esterase-cholesterol oxidase.
The preparation method of micro-fluidic refill sheet may further comprise the steps:
1) by photoetching technique, manufactures the chip duct at photoetching paper;
2) will carry out the lamination assembling with the filter paper in duct and double faced adhesive tape, in the assembling process in the hole fiberfill fibers element powder/aqueous mixtures;
3) with glucose oxidase, urate oxidase, GPO, cholesterol esterase-cholesterol oxidase, peroxidase, 4-AA and N-ethyl-N-(3-sulfoacetic acid)-3 methoxybenzene amine aqueous solutions, join respectively assigned address A, B, C, the D zone of refill sheet, the lucifuge room temperature is air-dry.
The object of the present invention is achieved like this: specific oxidation enzyme (glucose oxidase, urate oxidase, GPO, cholesterol esterase-cholesterol oxidase) and peroxidase enzyme are fixed on the refill sheet, with the activity of Trehalosc protection enzyme.Glucose, uric acid, triglyceride and cholesterol are generating H under the oxidasic catalysis separately
2O
2, and H
2O
2Under 4-AA and the participation of N-ethyl-N-(3-sulfoacetic acid)-3 aminoanisoles, generate red quinone imines pigment again.But micro-fluidic refill sheet filtered sample, and realize that the shunting of sample detects.The overall process of three-dimensional microflow control refill piece making method may further comprise the steps:
1) covers the shading lid that designs passage at the filter paper that is brushed with photoresist, pass through uv-exposure, the regional photoresist generation cross-linking reaction of not covered by passage is removed by the washing of developer solution by the regional optical cement that passage is covered, and makes this zone recover water wettability;
2) according to the design of mask upper channel, there is the common double faced adhesive tape of parchamyn paper to punch on the two sides.All double faced adhesive tapes and paper cut into same size before assembling so that assembling.Flow direction according to solution begins assembling from entrance, throws off a parchamyn paper, the relevant position firm pasting of aligned inlet passage.In the hole, fill an amount of cellulose powder/aqueous mixtures, tear again another layer parchamyn paper off, cling with one deck refill sheet that is close to.Paste according to the process that refill sheet and double faced adhesive tape replace repeatedly, will stick with one deck refill sheet that detects reagent at last;
3) adopt phosphate buffer to prepare respectively oxidase (glucose oxidase, urate oxidase, GPO, cholesterol esterase-cholesterol oxidase) solution and the superoxide enzyme-substrate chromophoric solution that contains trehalose.Mentioned solution is added on the refill sheet successively, and the lucifuge room temperature is air-dry.
Different according to adding oxidation enzyme class, carry out figure notation at refill sheet pro and con and (add region, the hole reference numerals " A " of glucose oxidase; Add region, the hole reference numerals " B " of urate oxidase; Add region, the hole reference numerals " C " of GPO; Add region, the hole reference numerals " D " of cholesterol esterase-cholesterol oxidase).
When 4) detecting, sample drop is added in the well in refill sheet front, as containing determinand in the sample, then the colour developing hole at the back side presents redness.
, reagent mobile by capillary fringe hydrodynamic body adsorbs and filtered sample, does not need foreign aid's equipment such as expensive pump or pressure control valve; Sample drop is added in the application of sample face, and as containing determinand in the sample, then the colour developing hole at the back side presents redness.
Description of drawings
The structural map of Fig. 1 three-dimensional microflow control refill sheet;
Synoptic diagram is detected in the front of Fig. 2 three-dimensional microflow control refill sheet;
The reverse side of Fig. 3 three-dimensional microflow control refill sheet detects synoptic diagram;
Wherein: A glucose detection district, B uric acid detection zone, C triglyceride detection zone, D cholesterol detection district, 1 hole, 2 adhesive tapes, 3 light-sensitive emulsions, 4 filter paper, 5 wells, 6 colour developing holes.
Specific embodiment
Embodiment 1
Concrete preparation, assembling, testing process are as follows:
The preparation process of photoetching microchannel is as follows among 1 the present invention:
At TX609 paper brushing SU-8 photoresist, 130 ℃ of baking 10min on electric hot plate, room temperature is cooled off, and designs the mask of pattern in the covering, uv-exposure 14s, 130 ℃ of baking 10min on electric hot plate, room temperature cooling.The regional photoresist generation cross-linking reaction of not covered by passage is removed by the acetone washing by the regional photoresist that passage is covered, and adopts the oxygen plasma cleaning machine to clean 3s, makes passage area recover water wettability.
The assembling process of three-dimensional microflow control refill sheet is as follows among 2 the present invention:
According to the design of mask upper channel, there is the common double faced adhesive tape of parchamyn paper to punch on the two sides.All double faced adhesive tapes and paper cut into same size before assembling so that assembling.Flow direction according to solution begins assembling from entrance, throws off a parchamyn paper, the relevant position firm pasting of aligned inlet passage.In the hole, fill an amount of cellulose powder/aqueous mixtures, tear again another layer parchamyn paper off, cling with one deck refill sheet that is close to.Paste according to the process that refill sheet and double faced adhesive tape replace repeatedly, will stick with one deck refill sheet that detects reagent at last.
The detection system building process of three-dimensional microflow control refill sheet is as follows among 3 the present invention:
Prepare respectively phosphate-buffered liquor (the various oxidase 120U/mL of glucose oxidase, urate oxidase, GPO, cholesterol esterase-cholesterol oxidase, 0.3mol/L trehalose), superoxide enzyme-substrate chromophoric solution (horseradish peroxidase 30U/mL, 4-AA 0.82mmol/L, N-ethyl-N-(3-sulfoacetic acid)-3 aminoanisole 14mmol/L).Each 2 μ L of above-mentioned two kinds of solution are added on the refill sheet successively, and the lucifuge room temperature is air-dry.
4 micro-fluidic refill sheet using method are as follows:
5～10 μ L liquid to be measured is added in the well in micro-fluidic refill sheet front, leave standstill 5min.If only contain glucose in the liquid to be measured, colour developing hole, so micro-fluidic refill sheet reverse side A district presents redness; If only contain uric acid in the liquid to be measured, colour developing hole, so micro-fluidic refill sheet reverse side B district presents redness; If only contain triglyceride in the liquid to be measured, colour developing hole, so micro-fluidic refill sheet reverse side C district presents redness; If only contain cholesterol in the liquid to be measured, colour developing hole, so micro-fluidic refill sheet reverse side D district presents redness; If contain simultaneously glucose and uric acid in the liquid to be measured, so micro-fluidic refill sheet reverse side A and colour developing hole, B district present redness; In like manner, if contain simultaneously glucose, uric acid, triglyceride and cholesterol in the liquid to be measured, the colour developing Kong Jun in so micro-fluidic refill sheet reverse side A, B, C, four districts of D presents redness.
Claims (3)
1. detect simultaneously the micro-fluidic refill sheet of glucose, uric acid, triglyceride and cholesterol, it is characterized in that: the refill sheet includes specific oxidation enzyme, peroxidase, 4-AA and N-ethyl-N-(3-sulfoacetic acid)-3 aminoanisoles by the three-dimensional micropore road system that photoetching filter paper and the stack of punching double faced adhesive tape form.
2. micro-fluidic refill sheet as claimed in claim 1, it is characterized in that: the specific oxidation enzyme is glucose oxidase, urate oxidase, GPO and cholesterol esterase-cholesterol oxidase.
3. the preparation method of micro-fluidic refill sheet as claimed in claim 1 is characterized in that may further comprise the steps:
1) by photoetching technique, manufactures the chip duct at photoetching paper;
2) will carry out the lamination assembling with the filter paper in duct and double faced adhesive tape, in the assembling process in the hole fiberfill fibers element powder/aqueous mixtures;
3) with glucose oxidase, urate oxidase, GPO, cholesterol esterase-cholesterol oxidase, peroxidase, 4-AA and N-ethyl-N-(3-sulfoacetic acid)-3 methoxybenzene amine aqueous solutions, join respectively assigned address A, B, C, the D zone of refill sheet, the lucifuge room temperature is air-dry.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011102702132A CN102998439A (en) | 2011-09-14 | 2011-09-14 | Micro-fluidic paper chip for simultaneously detecting glucose, uric acid, triglycerides and cholesterols, and its manufacturing method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011102702132A CN102998439A (en) | 2011-09-14 | 2011-09-14 | Micro-fluidic paper chip for simultaneously detecting glucose, uric acid, triglycerides and cholesterols, and its manufacturing method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102998439A true CN102998439A (en) | 2013-03-27 |
Family
ID=47927276
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011102702132A Pending CN102998439A (en) | 2011-09-14 | 2011-09-14 | Micro-fluidic paper chip for simultaneously detecting glucose, uric acid, triglycerides and cholesterols, and its manufacturing method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102998439A (en) |
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104502516A (en) * | 2015-01-27 | 2015-04-08 | 天津出入境检验检疫局工业产品安全技术中心 | Micro-fluidic refractive index detection method used for polymerized triglyceride |
| CN106018522A (en) * | 2016-07-25 | 2016-10-12 | 济南大学 | Preparation of photo-induced chemical paper chip for detecting expression of polysaccharides on surfaces of cancer cells |
| CN106485250A (en) * | 2016-09-09 | 2017-03-08 | 华东理工大学 | A kind of recognition methods of paper chip gray value and its application |
| CN108273573A (en) * | 2017-12-29 | 2018-07-13 | 哈尔滨工业大学 | It is a kind of can a step realize ELISA immune responses three-dimensional paper chip and preparation method thereof |
| CN108745428A (en) * | 2018-06-05 | 2018-11-06 | 合肥工业大学 | A kind of multichannel three-dimensional microflow control paper chip and preparation method thereof |
| CN109012770A (en) * | 2018-07-12 | 2018-12-18 | 中国科学院成都生物研究所 | Multi-ply paper chip structure, manufacturing equipment and method and fluid interlayer current method |
| CN109916894A (en) * | 2019-04-12 | 2019-06-21 | 吉林省汇酉生物技术股份有限公司 | A kind of dry chemistry reagent piece and preparation method thereof quantitative determining concentration of glucose |
| CN109916890A (en) * | 2019-04-12 | 2019-06-21 | 吉林省汇酉生物技术股份有限公司 | A kind of dry chemistry reagent piece and preparation method thereof quantitative determining total cholesterol concentration |
| CN110646410A (en) * | 2019-07-23 | 2020-01-03 | 太原理工大学 | Cardiovascular and cerebrovascular disease multi-index joint diagnosis method based on combination of portable scanner and microfluidic filter paper chip |
| CN111351925A (en) * | 2019-07-29 | 2020-06-30 | 中南大学 | Paper chip capable of simultaneously detecting various aminoglycoside antibiotics, preparation and application thereof |
| CN111781264A (en) * | 2020-07-15 | 2020-10-16 | 桂林电子科技大学 | Preparation method of PtNPs-based 3D paper-based electrochemical glucose sensor |
| CN113959997A (en) * | 2021-09-29 | 2022-01-21 | 中南大学 | 3D folding ratio fluorescence microfluidic device and method for simultaneously detecting alkaline phosphatase and butyrylcholinesterase |
| CN114317678A (en) * | 2021-12-31 | 2022-04-12 | 港龙生物技术（深圳）有限公司 | Biological paper chip, high-flux multi-connection detection microporous plate device, preparation method and kit for multi-connection detection of vaginitis |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1446925A (en) * | 2003-03-24 | 2003-10-08 | 肖洪武 | Single stable colorimetric reagent of enzyme in liquid state and its application |
| CN1950700A (en) * | 2004-04-30 | 2007-04-18 | 爱科来株式会社 | Sample analyzing tool |
| CN101025415A (en) * | 2006-02-23 | 2007-08-29 | 顺德职业技术学院 | Test paper for detecting uric acid content range in urine |
| WO2008043543A2 (en) * | 2006-10-10 | 2008-04-17 | Miacom Diagnostics Gmbh | Nucleic acid beacons for fluorescent in-situ hybridisation and chip technology |
| EP2179291A2 (en) * | 2007-07-13 | 2010-04-28 | Prometheus Laboratories, Inc. | Drug selection for lung cancer therapy using antibody-based arrays |
| CN101762628A (en) * | 2009-10-10 | 2010-06-30 | 张祥成 | Dry enzyme method biochip/test strip |
-
2011
- 2011-09-14 CN CN2011102702132A patent/CN102998439A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1446925A (en) * | 2003-03-24 | 2003-10-08 | 肖洪武 | Single stable colorimetric reagent of enzyme in liquid state and its application |
| CN1950700A (en) * | 2004-04-30 | 2007-04-18 | 爱科来株式会社 | Sample analyzing tool |
| CN101025415A (en) * | 2006-02-23 | 2007-08-29 | 顺德职业技术学院 | Test paper for detecting uric acid content range in urine |
| WO2008043543A2 (en) * | 2006-10-10 | 2008-04-17 | Miacom Diagnostics Gmbh | Nucleic acid beacons for fluorescent in-situ hybridisation and chip technology |
| EP2179291A2 (en) * | 2007-07-13 | 2010-04-28 | Prometheus Laboratories, Inc. | Drug selection for lung cancer therapy using antibody-based arrays |
| CN101762628A (en) * | 2009-10-10 | 2010-06-30 | 张祥成 | Dry enzyme method biochip/test strip |
Non-Patent Citations (1)
| Title |
|---|
| 王方方等: "纸芯片制作及其在化学发光法检测葡萄糖和尿酸中的应用", 《分析科学学报》 * |
Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104502516A (en) * | 2015-01-27 | 2015-04-08 | 天津出入境检验检疫局工业产品安全技术中心 | Micro-fluidic refractive index detection method used for polymerized triglyceride |
| CN106018522A (en) * | 2016-07-25 | 2016-10-12 | 济南大学 | Preparation of photo-induced chemical paper chip for detecting expression of polysaccharides on surfaces of cancer cells |
| CN106485250A (en) * | 2016-09-09 | 2017-03-08 | 华东理工大学 | A kind of recognition methods of paper chip gray value and its application |
| CN108273573A (en) * | 2017-12-29 | 2018-07-13 | 哈尔滨工业大学 | It is a kind of can a step realize ELISA immune responses three-dimensional paper chip and preparation method thereof |
| CN108745428B (en) * | 2018-06-05 | 2020-11-27 | 合肥工业大学 | Multi-channel three-dimensional microfluidic paper chip and preparation method thereof |
| CN108745428A (en) * | 2018-06-05 | 2018-11-06 | 合肥工业大学 | A kind of multichannel three-dimensional microflow control paper chip and preparation method thereof |
| CN109012770B (en) * | 2018-07-12 | 2021-05-14 | 中国科学院成都生物研究所 | Multilayer paper chip structure, manufacturing apparatus and method, and fluid interlayer flow method |
| CN109012770A (en) * | 2018-07-12 | 2018-12-18 | 中国科学院成都生物研究所 | Multi-ply paper chip structure, manufacturing equipment and method and fluid interlayer current method |
| CN109916890A (en) * | 2019-04-12 | 2019-06-21 | 吉林省汇酉生物技术股份有限公司 | A kind of dry chemistry reagent piece and preparation method thereof quantitative determining total cholesterol concentration |
| CN109916894A (en) * | 2019-04-12 | 2019-06-21 | 吉林省汇酉生物技术股份有限公司 | A kind of dry chemistry reagent piece and preparation method thereof quantitative determining concentration of glucose |
| CN110646410A (en) * | 2019-07-23 | 2020-01-03 | 太原理工大学 | Cardiovascular and cerebrovascular disease multi-index joint diagnosis method based on combination of portable scanner and microfluidic filter paper chip |
| CN111351925A (en) * | 2019-07-29 | 2020-06-30 | 中南大学 | Paper chip capable of simultaneously detecting various aminoglycoside antibiotics, preparation and application thereof |
| CN111781264A (en) * | 2020-07-15 | 2020-10-16 | 桂林电子科技大学 | Preparation method of PtNPs-based 3D paper-based electrochemical glucose sensor |
| CN113959997A (en) * | 2021-09-29 | 2022-01-21 | 中南大学 | 3D folding ratio fluorescence microfluidic device and method for simultaneously detecting alkaline phosphatase and butyrylcholinesterase |
| CN114317678A (en) * | 2021-12-31 | 2022-04-12 | 港龙生物技术（深圳）有限公司 | Biological paper chip, high-flux multi-connection detection microporous plate device, preparation method and kit for multi-connection detection of vaginitis |
| CN114317678B (en) * | 2021-12-31 | 2023-11-10 | 港龙生物技术（深圳）有限公司 | Biological paper chip, high-throughput multi-detection microplate device, preparation method and kit for vaginal inflammation multi-detection |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102998439A (en) | Micro-fluidic paper chip for simultaneously detecting glucose, uric acid, triglycerides and cholesterols, and its manufacturing method | |
| Van Hoovels et al. | Can wearable sweat lactate sensors contribute to sports physiology? | |
| CN110573063B (en) | Colorimetric sensor for non-invasive screening of glucose in sweat of prodromal and type 2 diabetics | |
| US20050175505A1 (en) | Personal monitor to detect exposure to toxic agents | |
| US11612356B2 (en) | On-diaper body fluid screening device and associated method | |
| CN110361529A (en) | A kind of reagent card, detection method and application | |
| WO2010090810A2 (en) | Diagnostic device and method | |
| CN107217001A (en) | A kind of detection of biological samples device and the gene tester based on the device | |
| Brazaca et al. | The use of biological fluids in microfluidic paper-based analytical devices (μPADs): Recent advances, challenges and future perspectives | |
| CN107782906A (en) | A kind of detection means and detection method | |
| CN201955341U (en) | Diagnostic instrument for oral liquid | |
| WO2018165180A1 (en) | Colorometric sensor for the non-invasive screening of glucose in sweat in pre and type 2 diabetes | |
| EP3575780A1 (en) | Fecal occult blood test pad | |
| ES2430072T3 (en) | Test element for the analysis of a body fluid | |
| KR20110064906A (en) | Diagnostic kit for respiratory disease | |
| Dixon et al. | The diagnostic potential of microneedles in infectious diseases | |
| CN102089441B (en) | Enzymatic analytical membrane, test device and method | |
| KR20230065972A (en) | Devices for Analyzing Peritoneal Dialysate | |
| CN207300865U (en) | The chip of blood glucose and blood fat is detected at the same time | |
| CN107384772A (en) | One kind can scale paper substrate detection chip and its manufacture method | |
| US20080237038A1 (en) | Biostrip | |
| CN200975990Y (en) | Liquid detection test piece | |
| CN202757874U (en) | Body fluid sample special quantitative detection slide | |
| CN204594918U (en) | Anti-interference sensing electrode | |
| TW201623955A (en) | A biosensor with structure of micro chamber |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20130327 |