CN1912622A - Chemiluminescence investigating method of glucose in body fluid - Google Patents
Chemiluminescence investigating method of glucose in body fluid Download PDFInfo
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- CN1912622A CN1912622A CN 200610114973 CN200610114973A CN1912622A CN 1912622 A CN1912622 A CN 1912622A CN 200610114973 CN200610114973 CN 200610114973 CN 200610114973 A CN200610114973 A CN 200610114973A CN 1912622 A CN1912622 A CN 1912622A
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Abstract
A method for determining glucose in body fluid utilizing chemiluminescence includes oxidizing glucose by glucose oxidase to generate perhydrol, reacting on peroxidase with perhydrol to make chemiluminescent matter be oxidized for lightening and presenting light signal size to be positive correlation with concentration of glucose, recording this light signal in order to estimate out glucose concentration, using known concentration of glucose and measured light signal to work out dosage-reaction curve for calculating out glucose content in unknown sample by utilizing said curve.
Description
Technical field: the present invention relates to a kind of medical agent detection method, particularly the chemical luminescent detecting method of glucose in the body fluid.
Background technology: the ultramicro-determination method of utilizing modern biotechnology to develop has promoted the fast development of the every clinical and research work in biological and each field of medical science, greatly for outstanding contribution has been made in the mankind's cause of science and social progress.
Last century, the seventies was at first delivered the application luminol by Arakawa---the chemiluminescence reaction of peroxidase is carried out the technology that enzyme exempts to analyze, this technology since sensitivity up to 10
-18Mol can fast measuring go out the result in 20 minutes, can easy realization full automation, be valid up to more than 1 year, and "dead" and teratogen is beneficial to environmental protection and is well received, and is the detection technique the most rapidly of development at present.
At present, chemiluminescence has obtained quite universal degree in immunoassay, developed into the routine prison bed detection method that can detect multiple bioactivator, from immune response mode branch, this method has two kinds of fundamental types, be chemiluminescence immune assay (chemiluminescence immunoassay, CLIA) and immunochemiluminometry (Immunochemiluminometricassay, ICMA); From illumination mode, the one, with the direct mark of luminescent substance on antibody or antigen, after the immune response, and luminous, another kind is with enzymic-labelled antibody or antigen under the effect of incipient reagent, after the immune response, add chemical luminous substrate, under the enzyme caloytic action and luminous.
Though this method has outstanding advantage, only limit to immunoassay at present, and still blank in biochemical analysis.
The method of glucose is biological enzyme in the mensuration body fluid that now generally uses, this method is to utilize biology enzyme to make glucose emit hydrogen peroxide, the latter urges down at enzyme look source material is oxidized to colored substance, judges the concentration of glucose in the biological fluid by shade.The weak point of this method mainly contains:
1. the range of linearity is narrow.Because detection is so responsive unlike chemiluminescence to change in color resolution, the range of linearity can not satisfy the needs of clinical detection fully, need measure after diluting the sample of high concentration, brings certain inconvenience.
2. sensitivity is not as chemiluminescence.Enzyme process is the depth than color, and chemiluminescence is to measure light signal, and sensitivity is high.
3. can only use instrumentation.Have plenty of the speed of measuring enzymatic reaction, very strict to time requirement, the equal difficulty of craft or semi-automatic operation reaches requirement, can only use fully-automatic equipment to carry out.
4. full automatic biochemical apparatus costliness.The full automatic biochemical apparatus that uses at present almost is import entirely, and equipment is very expensive, mostly hundreds of thousands arrive millions of between, middle and small hospital is difficult to bear, universal use is restricted.
5. some the look source material in the enzyme process is harmful to, and easily causes environmental pollution, and is not fool proof.
Summary of the invention: technical matters to be solved by this invention is a kind of chemical luminescent detecting method that can overcome glucose in above-mentioned defective, highly sensitive, the body fluid that the range of linearity is wide, convenient, safe that provides.Solve this technical problem the main technical schemes of employing: the chemical luminescent detecting method of glucose in the body fluid is characterized in that glucose oxidase makes glucose oxidase and generates hydrogen peroxide; Hydrogen peroxide makes the chemiluminescent substance oxidation and luminous under the effect of peroxidase; The size of light signal and the concentration of glucose are proportionate, and promptly the big more light signal that sends of concentration of glucose is strong more; Write down this light signal and can infer the concentration of glucose; With the glucose of concentration known and the light signal of measuring, make dose-response curve; The glucose content of unknown sample can come out from this curve calculating; Described chemiluminescent substance is a luminol; Described glucose can be the glucose in the blood; Described glucose can be the glucose in the blood plasma; Described glucose can be the glucose in the serum; Described glucose can be the glucose in the cerebrospinal fluid; Described glucose can be the glucose in the urine.The present invention compared with prior art has following advantage: the method is characterized in that: replace the colour developing thing with chemiluminescent substance, reach sensitiveer, more stable, wide ranges, safer purpose.The commercially available reagent box that can prepare corresponding quantitative measurement glucose with this method.Concrete advantage is: 1. sensitivity is higher.Classic method is to detect shade, and chemiluminescence is the photometry signal, photon counting one by one, and the minimum of being surveyed is littler, thereby sensitivity is higher.2. the range of linearity is wide.The resolution of the depth of survey color is more much lower than the precision of photon counting, and chemiluminescent sensing range is up to 10
53. safe and applicable.Chemiluminescent substance such as luminol etc. are very safe, and whole testing process does not have nuisance and occurs, and are that detection or processing of waste are all very safe, help environmental protection.4. be convenient to popularize.The kit that the present invention is prepared can be used for all automatic measurement, also can be used for craft or semi-automatic measuring, and the medical treatment of suitable large, medium and small each level and research institution use.5. huge social benefit.With chemiluminescence determination biochemical indicator or immune substance, can no longer need the biochemical instruments of import costliness together with a Chemiluminescence Apparatus, save substantial contribution, not only reduced cost, medical institutions, patient and society are all benefited.
Embodiment: now illustrate embodiments of the invention.Glucose oxidase makes glucose oxidase and generates hydrogen peroxide; Hydrogen peroxide makes the chemiluminescent substance oxidation and luminous under the effect of peroxidase; The size of light signal and the concentration of glucose are proportionate, and promptly the big more light signal that sends of concentration of glucose is strong more; Write down this light signal and can infer the concentration of glucose; With the glucose of concentration known and the light signal of measuring, make dose-response curve; The glucose content of unknown sample can come out from this curve calculating; Described chemiluminescent substance is a luminol; Described glucose can be the glucose in the blood; Described glucose can be the glucose in the blood plasma; Described glucose can be the glucose in the serum; Described glucose can be the glucose in the cerebrospinal fluid; Described glucose can be the glucose in the urine.This is the chemical luminescent detecting method of glucose in the body fluid of the present invention.This method can be used for measuring the glucose content in blood, blood plasma, serum, cerebrospinal fluid or the urine.
One, reagent preparation:
0.1MpH7.0 PB
GOD+HRP liquid.
Glucose standard: single or series concentration;
Substrate solution
Two, mensuration program:
According to the form below carries out, and application of sample unit is μ l.
| Standard | Sample | |
| Standard | 20 | |
| Sample | 20 | |
| Enzyme liquid | 50 | 50 |
| Substrate solution | 50 | 50 |
| Mixing, measure R LU behind room temperature or the 37 ℃ of incubations | ||
Three, make dose-response curve with the proper method processing, RLU value is per sample obtained the concentration of its serum glucose from this curve.Or directly obtain by formula:
Concentration of glucose=(sample RLU/ standard RLU) * normal concentration
Four, also available double reagent is measured.
Sensitivity of the present invention is higher: because classic method is to detect shade, and the present invention is the photometry signal, photon counting one by one, and the minimum of being surveyed is littler, thereby sensitivity is higher.The range of linearity is wide: this is that chemiluminescence detection scope of the present invention is up to 10 because the resolution of the depth of traditional survey color is more much lower than the precision of photon counting
5Safe and applicable: chemiluminescent substance of the present invention such as luminol etc. are very safe, and whole testing process does not have nuisance and occurs, and are that detection or processing of waste are all very safe, help environmental protection.Be convenient to popularize: the kit that the present invention is prepared, can be used for all automatic measurement, also can be used for craft or semi-automatic measuring, the medical treatment of suitable large, medium and small each level and research institution use.Social benefit is huge: measure biochemical indicator or immune substance with chemical luminescent detecting method of the present invention, can no longer need the biochemical instruments of import costliness together with a Chemiluminescence Apparatus, save substantial contribution, not only reduced cost, medical institutions, patient and society have all been benefited.The inventive method is simple, and cost is low, is suitable for large, medium and small medical institutions wide popularization and application.
Claims (7)
1, the chemical luminescent detecting method of glucose in the body fluid is characterized in that glucose oxidase makes glucose oxidase and generates hydrogen peroxide; Hydrogen peroxide makes the chemiluminescent substance oxidation and luminous under the effect of peroxidase; The size of light signal and the concentration of glucose are proportionate, and promptly the big more light signal that sends of concentration of glucose is strong more; Write down this light signal and can infer the concentration of glucose; With the glucose of concentration known and the light signal of measuring, make dose-response curve; The glucose content of unknown sample can come out from this curve calculating.
2, the chemical luminescent detecting method of glucose in the body fluid according to claim 1 is characterized in that described chemiluminescent substance is a luminol.
3, the chemical luminescent detecting method of glucose in the body fluid according to claim 1 is characterized in that described glucose can be the glucose in the blood.
4, the chemical luminescent detecting method of glucose in the body fluid according to claim 1 is characterized in that described glucose can be the glucose in the blood plasma.
5, the chemical luminescent detecting method of glucose in the body fluid according to claim 1 is characterized in that described glucose can be the glucose in the serum.
6, the chemical luminescent detecting method of glucose in the body fluid according to claim 1 is characterized in that described glucose can be the glucose in the cerebrospinal fluid.
7, the chemical luminescent detecting method of glucose in the body fluid according to claim 1 is characterized in that described glucose can be the glucose in the urine.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200610114973 CN1912622A (en) | 2006-08-10 | 2006-08-10 | Chemiluminescence investigating method of glucose in body fluid |
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| CN 200610114973 CN1912622A (en) | 2006-08-10 | 2006-08-10 | Chemiluminescence investigating method of glucose in body fluid |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102053085B (en) * | 2009-10-28 | 2013-03-20 | 中国科学院高能物理研究所 | Method for detecting glucose by ferroferric oxide nano particle catalytic chemiluminescence |
| CN104990913A (en) * | 2015-05-29 | 2015-10-21 | 南开大学 | Gold nanoparticle growth based method for detection of glucose |
| CN105062490A (en) * | 2015-07-23 | 2015-11-18 | 成都理工大学 | Preparation method for integrated bienzyme-quantum dots used for chemiluminescent resonance energy transfer |
| CN106546578A (en) * | 2016-10-28 | 2017-03-29 | 华南师范大学 | The gravity on content of wax dam/capillary flow cloth chip and its application in glucose sensing |
| CN110231486A (en) * | 2019-06-28 | 2019-09-13 | 吉林大学 | A kind of detection method of glucose |
-
2006
- 2006-08-10 CN CN 200610114973 patent/CN1912622A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102053085B (en) * | 2009-10-28 | 2013-03-20 | 中国科学院高能物理研究所 | Method for detecting glucose by ferroferric oxide nano particle catalytic chemiluminescence |
| CN104990913A (en) * | 2015-05-29 | 2015-10-21 | 南开大学 | Gold nanoparticle growth based method for detection of glucose |
| CN104990913B (en) * | 2015-05-29 | 2017-12-15 | 南开大学 | A kind of method based on gold nano grain detection of the growth glucose |
| CN105062490A (en) * | 2015-07-23 | 2015-11-18 | 成都理工大学 | Preparation method for integrated bienzyme-quantum dots used for chemiluminescent resonance energy transfer |
| CN106546578A (en) * | 2016-10-28 | 2017-03-29 | 华南师范大学 | The gravity on content of wax dam/capillary flow cloth chip and its application in glucose sensing |
| CN106546578B (en) * | 2016-10-28 | 2019-08-30 | 华南师范大学 | The gravity on content of wax dam/capillary flow cloth chip and its application in glucose sensing |
| CN110231486A (en) * | 2019-06-28 | 2019-09-13 | 吉林大学 | A kind of detection method of glucose |
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