CN110346570A - Detect the colloidal gold kit and method of diabetic nephropathy biomarker - Google Patents
Detect the colloidal gold kit and method of diabetic nephropathy biomarker Download PDFInfo
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- CN110346570A CN110346570A CN201810301654.6A CN201810301654A CN110346570A CN 110346570 A CN110346570 A CN 110346570A CN 201810301654 A CN201810301654 A CN 201810301654A CN 110346570 A CN110346570 A CN 110346570A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/04—Endocrine or metabolic disorders
- G01N2800/042—Disorders of carbohydrate metabolism, e.g. diabetes, glucose metabolism
Abstract
The present invention provides a kind of colloidal gold kit and method for detecting diabetic nephropathy biomarker, which includes sample pad, gold-labelled pad, nitrocellulose filter, water absorption pad and supporting pad.Nitrocellulose filter sets gradually the first detection reagent article, the second detection reagent article, third detection reagent article, the 4th detection reagent article and capture reagent strip.First detection reagent item is coated with anti-type Ⅳ collagen protein monoclonal antibody, the anti-sufficient glycocalicin monoclonal antibody of second detection reagent item coating, third detection reagent item is coated with anti-neutrophil gelatinase-associated lipocalin monoclonal antibody, and the 4th detection reagent article is coated with anti-liver type fatty acid binding protein monoclonal antibody；Capture the secondary antibody that reagent strip is coated in conjunction with monoclonal antibody coated in gold-labelled pad.Colloidal gold kit of the present invention realizes a variety of biomarkers of simultaneous quantitative detection early diabetic nephropathy, saves the cost of detection early diabetic nephropathy, improves detection efficiency.
Description
Technical field
The invention belongs to biomarker detection field more particularly to a kind of glue for detecting diabetic nephropathy biomarker
Body gold reagent box and method.
Background technique
Diabetic nephropathy is one of chronic microvascular complication of diabetes most serious, and finally causes terminal kidney failure,
It is the main reason for diabetic is dead.However, diabetic nephropathy early stage is difficult to find, and clinical diabetes diagnosis of nephropathy is yellow
Goldstandard creatinine and albuminuria can only also reflect that nephrolithotomy venereal disease becomes indirectly, and early stage sugar can not be diagnosed during normal albuminuria
Urinate sick nephrosis.Usually when clinical definite, Diabetic Nephropathy patients miss optimal therapic opportunity, and disease is caused sharply to dislike
Change, it is irreversible.Therefore, for the early discovery of diabetic nephropathy earlier damage biomarker, early intervention, there is important reality
Meaning.
It has now been found that many early diabetic nephropathies relevant biomarkers, some biomarkers can be
This stage is discharged into urine, and the height of protein classes and content in urine directly reflects urinary system, especially kidney
Health status, can predict diabetic nephropathy there is a situation where, development and prognosis.Such as sufficient glycocalicin
(podocalyxin), it is the early stage biomarker of Podocytes in Renal Tissue, and is positively correlated with the development of disease；Type Ⅳ collagen albumen
(Collagen IV) is played an important role in terms of maintenance cell basilar memebrane correctly assembly；Nephrosis albumen (Nephrin),
It splits in the assembly of sky diaphragm and plays an important role；Liver type fatty acid binding protein (L-FABP) is deep cell damage
The biomarker of wound can predict the generation of diabetic nephropathy；Neutrophil gelatinase-associated lipocalin
(NGAL), cystatin C (cystatinC), nerve growth factor -1 (Netrin-1) are the biological markers of renal tubule Urgent Management of Trauma of Proximal
Object.To these, the biomarker closely related with disease development is detected, and helps to understand diabetic nephropathy
And development, effectively improve the predictive ability to renal complications.
Common method is using Enzyme-multiplied immune technique in the detection device of detection early diabetic nephropathy at present
(ELISA), technical disadvantages are as follows: detection device volume is big, at high cost；Detection time is long, repeatability is bad, is not suitable for clinic
Rapid screening.However, being there is no at present using a variety of biomarkers while the equipment that carries out quantitative detection, therefore can not achieve
Simultaneous quantitative detects a variety of biomarkers of early diabetic nephropathy, to can not mention for early diabetic nephropathy clinical diagnosis
For foundation.
Summary of the invention
The main purpose of the present invention is to provide a kind of colloidal gold kit for detecting diabetic nephropathy biomarker, purports
Solving the problems, such as that current detection device can not the simultaneous quantitative detection a variety of biomarkers of early diabetic nephropathy.
To achieve the above object, the present invention provides a kind of colloid gold reagents for detecting diabetic nephropathy biomarker
Box, the colloidal gold kit include sample pad, gold-labelled pad, nitrocellulose filter, water absorption pad and supporting pad, wherein described
Sample pad, gold-labelled pad, nitrocellulose filter, the overlapped one section of preset length of water absorption pad head and the tail, on the nitrocellulose filter
Set gradually the first detection reagent article, the second detection reagent article, third detection reagent article, the 4th detection reagent article and capture examination
Agent item, in which:
The sample pad includes type Ⅳ collagen protein standard substance, sufficient glycocalicin standard items, neutrophil leucocyte gelatinase phase
Close lipocalin protein standard items and liver type fatty acid binding protein standard items；
Type Ⅳ collagen protein monoclonal antibody, sufficient glycocalicin monoclonal antibody, neutral grain are coated in the gold-labelled pad
Cell gelatinase associated lipocalin monoclonal antibody and liver type fatty acid binding protein monoclonal antibody；
The first detection reagent item is coated with anti-type Ⅳ collagen protein monoclonal antibody, the second detection reagent item packet
There is anti-sufficient glycocalicin monoclonal antibody, the third detection reagent item is coated with anti-neutrophil leucocyte gelatinase related lipid
Transporter monoclonal antibody, the 4th detection reagent article are coated with anti-liver type fatty acid binding protein monoclonal antibody；
The capture reagent strip is coated with the secondary antibody in conjunction with monoclonal antibody coated in the gold-labelled pad.
Preferably, first detection reagent article, the second detection reagent article, third detection reagent article, the 4th detection reagent
Item and capture reagent strip further include the indicator for having the biomarker for quantitative biological sample liquid.
Preferably, the indicator is enzyme, prothetic group, fluorescent material, bioluminescence substance or radioactive materials.
Preferably, first detection reagent article, the second detection reagent article, third detection reagent article, the 4th detection reagent
It is along the sample chromatography direction in sample pad and according to biological marker that putting in order for the nitrocellulose filter, which is arranged in, in item
Object molecular weight parallel arranged at equal intervals from big to small.
Preferably, the preset length is 1.5~2mm, and the secondary antibody is sheep anti-mouse igg or rabbit anti-mouse igg.
Preferably, the sample pad and gold-labelled pad are all made of glass fibre membrane or polyester film material is made.
Preferably, the supporting pad is a kind of rigid plastics bottom plate or PVC support baseboard.
Preferably, the water absorption pad is made of the water-absorption fiber or water-absorbing sponge material of water absorbing capacity, for adsorbing simultaneously
The biological sample liquid in the gold-labelled pad is drained, makes biological sample liquid by the gold-labelled pad by gold labeling antibody compound edge
Sample liquids chromatography direction be adsorbed on the nitrocellulose filter corresponding detection reagent position or capture reagent strip
Position.
The present invention also provides a kind of method using colloidal gold kit detection diabetic nephropathy biomarker, this method
Include the following steps: to obtain urine from subject as biological sample liquid；Biological sample liquid is poured into the sample-adding of gold-labelled pad
At position；The biomarker of biological sample liquid forms colloidal gold antibody compound in conjunction with the corresponding antibodies in gold-labelled pad；
Colloidal gold antibody compound with sample liquids on nitrocellulose membrane chromatographic flow, by the first detection reagent item, second detection
Reagent strip, third detection reagent article, the monoclonal antibody in the 4th detection reagent article combine；Color on judgement capture reagent strip
Whether change；If the color on capture reagent strip changes, by the first detection reagent item, the second detection reagent
Article, third detection reagent article and the color of the 4th detection reagent article and the standard items color of sample pad be compared and obtain biological mark
The content of will object.
Preferably, the detection method of the colloidal gold kit of the detection diabetic nephropathy biomarker further include as
Lower step: if the color on capture reagent strip changes, it is determined that the first detection reagent item of colloidal gold kit, second
Detection reagent article, third detection reagent article, the 4th detection reagent article have failed.
Compared to the prior art, the colloidal gold kit of detection diabetic nephropathy biomarker of the present invention is using upper
Technical solution is stated, following technical effect is reached: can be examined for a variety of early stage biomarkers of early diabetic nephropathy
It surveys, it is poor to solve the problems, such as that narrow and foresight is composed in single creature marker screening, testing cost can be saved, improve detection standard
True property.In addition, the colloidal gold kit can be used together while realizing a variety of biomarkers in conjunction with instrument
Quantitative detection provides foundation for early diabetic nephropathy clinical diagnosis, improves detection efficiency.
Detailed description of the invention
Fig. 1 is the structural representation of the colloidal gold kit preferred embodiment of present invention detection diabetic nephropathy biomarker
Figure；
Fig. 2 is the method preferred embodiment using colloidal gold kit of the present invention detection diabetic nephropathy biomarker
Flow diagram.
The embodiments will be further described with reference to the accompanying drawings for the realization, the function and the advantages of the object of the present invention.
Specific embodiment
Further to illustrate the technical means and efficacy of the invention taken to reach above-mentioned purpose, below in conjunction with attached drawing
And preferred embodiment, a specific embodiment of the invention, structure, feature and its effect are described in detail.It should be appreciated that this
Locate described specific embodiment to be only used to explain the present invention, be not intended to limit the present invention.
It should be noted that the colloidal gold kit that the present invention detects diabetic nephropathy biomarker is suitble to detection following
Four kinds of biomarkers provide auxiliary by detecting the content of these four biomarkers for diabetogenous nephrosis disease early diagnosis.
Sufficient glycocalicin (podocalyxin) is the transmembrane glycoprotein of O connection, is located at Renal Podocytes podocytic process teleblem area,
It is the main component of glomerular basement membrane electrostatic barrier, guarantees the integrality of glomerular filtration membrane electrostatic barrier, is filtered in kidney
It is played an important role in function.
Type Ⅳ collagen albumen (Collagen IV) is that the important set of glomerular basement membrane and extracellular matrix claims ingredient, small in kidney
It is played an important role in ball basilar memebrane filtration integrality.The stimulation of diabetes patient's hyperglycemia generates IV collagen, if increased
IV collagen is deposited on diabetes patient's renal glomerulus extracellular matrix, can generate dispersivity glomerulosclerosis；If deposition
Onto glomerular basement membrane or renal tubule, mesentery amplification, glomerulus interstitial and renal damage will lead to.
Liver type fatty acid binding protein (L-FABP) is expressed in renal proximal tubule cell, participates in renal tubular interstitium cell energy
Amount generates and metabolic process.In diabetic nephropathy concurrent process, in urine L-FABP expression and urine albumin concentration at
It is positively correlated, implies that L-FABP in urine is the biomarker of renal tubular interstitium cellular damage.
Neutrophil gelatinase-associated lipocalin (NGAL) is a kind of iron ion binding protein, in renal tubule
It is expressed in chrotoplast, participates in multiple biological function, such as Apoptosis, the innate immunity, kidney growth development.In diabetogenous nephrosis
Sick early stage, there is high expression in urine in NGAL, and the increase of NGAL expression quantity and the damage of renal tubule proximal tubule are proportional,
Imply that NGAL in urine is the biomarker of renal tubular interstitium cellular damage.
As shown in Figure 1, Fig. 1 is the colloidal gold kit preferred embodiment of present invention detection diabetic nephropathy biomarker
Structural schematic diagram.In the present embodiment, the colloidal gold kit include sample pad 1, gold-labelled pad 2, nitrocellulose filter 3,
Water absorption pad 4 and supporting pad 5.Wherein, the sample pad 1, gold-labelled pad 2, nitrocellulose filter 3,4 head and the tail of water absorption pad are overlapped
One section of preset length (such as 1.5~2mm) is simultaneously pasted on the supporting pad 5.The sample pad 1 includes sufficient glycocalicin standard
Product, type Ⅳ collagen protein standard substance, liver type fatty acid binding protein standard items and the delivery of neutrophil leucocyte gelatinase related lipid
Protein standard substance.
It is coated in the gold-labelled pad 2 in the gold-labelled pad and is coated with type Ⅳ collagen protein monoclonal antibody, sufficient glycocalicin
Monoclonal antibody, neutrophil gelatinase-associated lipocalin monoclonal antibody and liver type fatty acid binding protein Dan Ke
Grand antibody；
The first detection reagent item 31, the second detection reagent item 32, third inspection are disposed on the nitrocellulose filter 3
Test agent article 33, the 4th detection reagent article 34 and capture reagent strip 35.In the present embodiment, the first detection reagent item 31
It is coated with anti-type Ⅳ collagen protein monoclonal antibody, it is anti-that the second detection reagent item 32 is coated with anti-sufficient glycocalicin monoclonal
Body, the third detection reagent item 33 are coated with anti-neutrophil gelatinase-associated lipocalin monoclonal antibody, institute
It states the 4th detection reagent article 34 and is coated with anti-liver type fatty acid binding protein monoclonal antibody.The capture reagent strip 35 is coated
Antibody is the secondary antibody in conjunction with monoclonal antibody coated in gold-labelled pad 2, for example, sheep anti-mouse igg or rabbit anti-mouse igg.First inspection
Test agent article 31, the second detection reagent article 32, third detection reagent article 33, the 4th detection reagent article 34 and capture reagent strip 35
It further include having the indicator of the biomarker for quantitative biological sample liquid, such as enzyme, prothetic group, fluorescent material, biology are sent out
Stimulative substance or radioactive materials.One word of coating used in the embodiment of the present invention means the fixed meaning of non-specific adsorption,
The materials such as four kinds of monoclonal antibodies and reagent used in the embodiment of the present invention are common commercially available material unless otherwise indicated.
In the present embodiment, the first detection reagent item 31, the second detection reagent item 32, third detection reagent item 33,
4th detection reagent article 34 puts in order in the setting of the nitrocellulose filter 3 as along the sample chromatography side in sample pad 1
To 6 and according to biomarker molecular weight parallel arranged at equal intervals from big to small.For example, sufficient glycocalicin (podocalyxin),
Type Ⅳ collagen albumen (Collagen IV), liver type fatty acid binding protein (L-FABP) and neutrophil leucocyte gelatinase related lipid
Transporter (NGAL) is successively about 55KDa, 160KDa, 14KDa, 21KDa, therefore, nitrocellulose filter according to molecular weight
The first detection reagent article 31, the second detection reagent article 32, third detection reagent article 33 and the 4th detection reagent are disposed on 3
Coated anti-monoclonal antibody is from left to right followed successively by anti-type Ⅳ collagen protein monoclonal antibody, anti-sufficient glycocalicin to item 34 respectively
Monoclonal antibody, anti-neutrophil gelatinase-associated lipocalin monoclonal antibody, anti-liver type fatty acid binding protein
Monoclonal antibody.
In the present embodiment, the sample pad 1 and gold-labelled pad 2 are all made of glass fibre membrane or polyester film material is made.Institute
State the monoclonal antibody that label in gold-labelled pad 2 has biomarker.The first detection reagent item 31, the second detection reagent
Articles 32, third detection reagent article 33, coated antibody is mutual with the coated antibody of gold-labelled pad 2 respectively in the 4th detection reagent article 34
Match monoclonal antibody.The capture coated antibody of reagent strip 32 be it is a kind of can and gold-labelled pad 2 on the monoclonal antibody that marks
In conjunction with secondary antibody, for example, sheep anti-mouse igg or rabbit anti-mouse igg.The sample pad 1, gold-labelled pad 2, nitrocellulose filter 3, water absorption pad
The overlapped preset length (such as 1.5~2mm) of 4 head and the tail is simultaneously pasted on the supporting pad 5.The water absorption pad 4 is using water suction
The strong water-absorption fiber of ability or water-absorbing sponge material are made, and can have strong attract to flow to the sample liquids in the sample pad 1
Effect, i.e. sample liquids are by the gold-labelled pad 2 by gold labeling antibody-antigenic compound or gold labeling antibody compound along sample
The chromatography direction 6 of liquid is adsorbed on the nitrocellulose filter 3 corresponding detection reagent position or capture reagent strip position.
The supporting pad 5 is a kind of rigid plastics bottom plate or PVC support baseboard.
As shown in Fig. 2, Fig. 2 be using colloidal gold kit of the present invention detection diabetic nephropathy biomarker method compared with
The flow diagram of good embodiment.
Step S21 obtains urine as biological sample liquid from subject；Colloid gold reagent is stated utilizing using the present invention
When box detects the biomarker of Diabetic Nephropathy patients, tester obtains the urine conduct of Diabetic Nephropathy patients from subject
Biological sample liquid.
Step S22 pours into biological sample liquid at the load position of gold-labelled pad 2；Tester is by the biological sample of subject
Product liquid pours at the load position of gold-labelled pad 2.
Step S23, it is anti-that the biomarker of biological sample liquid forms colloidal gold in conjunction with the corresponding antibodies in gold-labelled pad 2
Nanocrystal composition；The urine of subject flows through gold-labelled pad 2, the relevant biomarker of early diabetic nephropathy and gold in Urine in Patients
The corresponding antibodies marked on pad 2 combine, and form colloidal gold antibody compound, i.e. antigenantibody complex.
Step S24, colloidal gold antibody compound with sample liquids on nitrocellulose membrane 3 chromatographic flow, by first inspection
Test agent article 31, the second detection reagent article 32, third detection reagent article 33, the monoclonal antibody knot in the 4th detection reagent article 34
It closes；Colloidal gold antibody compound with sample liquids on nitrocellulose membrane 3 chromatographic flow, when chromatography arrive detection reagent strip area
When, then the monoclonal antibody capture for being coated with marker in advance is trapped in the first detection reagent item 31, the second detection reagent item
32, third detection reagent article 33, in the 4th detection reagent article 34, when certain marker of capture is more, detects and try in a certain item
Color is deeper on agent item.
Whether step S25, the color captured on reagent strip 35 change；Tester estimates the color on capture reagent strip
Whether change.If the color on capture reagent strip 35 changes, step S26 is executed；If captured on reagent strip 35
Color there is no variation, execute step S27.In chromatography process, not by the first detection reagent item 31, the second detection reagent
Articles 32, third detection reagent article 33 and the colloidal gold antibody compound of the 4th detection reagent article 34 retention continue to move ahead, and work as arrival
35 region of reagent strip is captured, then can be identified by coated secondary antibody (such as sheep anti-mouse igg) in advance and is trapped in capture reagent strip 35
On, it is displayed in red.
Step S26, by the first detection reagent article 31, the second detection reagent article 32, third detection reagent article 33 and the 4th inspection
The color of test agent item 34 obtains the content of biomarker compared with the standard items color of sample pad.Tester is according to different inspections
The red depth carrys out the content of the unlike signal object in judgement sample on test agent item.If without containing certain mark in sample
Then color is not presented on corresponding detection reagent item in object, will be in corresponding detection reagent item if containing certain marker
The upper red that different depth degrees are presented.
Step S27 determines the various detection reagent items failure of colloidal gold kit.It, will if containing certain marker
The red of different depth degrees is presented on corresponding detection reagent item.It captures on reagent strip 35 if taken out of without red bar
It is existing, then illustrate the reagent failure of colloidal gold kit.
Urine (the biological sample liquid for stating colloidal gold kit detection early diabetic nephropathy patient is being utilized using the present invention
Body) when, the relevant biomarker of early diabetic nephropathy is formed in conjunction with the corresponding antibodies in gold-labelled pad 2 in Urine in Patients
Colloidal gold antibody compound (i.e. antigenantibody complex)；The compound is as sample liquids are on 3 upper layer of nitrocellulose membrane
Analysis flowing.When chromatography is to detection reagent strip area, then the monoclonal antibody capture for being coated with marker in advance is trapped in the
One detection reagent article 31, the second detection reagent article 32, third detection reagent article 33, in the 4th detection reagent article 34, capture certain
Kind marker is more, and color is deeper on a certain detection reagent item；Meanwhile in chromatography process, the colloidal gold that is not trapped
Antibody complex continues to move ahead, and captures 35 region of reagent strip when reaching, then can be by preparatory coated secondary antibody (such as sheep anti-mouse igg)
It identifies and is trapped on capture reagent strip 35, be displayed in red.Sample is judged according to the depth red on different detection reagent items 31
The content of unlike signal object in product.If not containing certain marker in sample, it is not on corresponding detection reagent item
The red of different depth degrees will be presented if containing certain marker in existing color on corresponding detection reagent item.Capture
If occurred without red stripes on reagent strip 35, illustrate the reagent failure of colloidal gold kit.Colloidal gold of the present invention
Kit have the advantages that detection be easy, be widely used, is easy to carry, screening it is comprehensive, and can be in conjunction with color collecting device
Quantitative detection while realizing multiple markers improves detection efficiency.
The above is only a preferred embodiment of the present invention, is not intended to limit the scope of the invention, all to utilize this hair
Equivalent structure made by bright specification and accompanying drawing content or equivalent function transformation, are applied directly or indirectly in other relevant skills
Art field, is included within the scope of the present invention.
Claims (10)
1. a kind of colloidal gold kit for detecting diabetic nephropathy biomarker, which is characterized in that the colloidal gold kit
Including sample pad, gold-labelled pad, nitrocellulose filter, water absorption pad and supporting pad, wherein the sample pad, gold-labelled pad, nitric acid are fine
Plain film, the overlapped one section of preset length of water absorption pad head and the tail are tieed up, sets gradually the first detection reagent on the nitrocellulose filter
Article, the second detection reagent article, third detection reagent article, the 4th detection reagent article and capture reagent strip, in which:
The sample pad includes type Ⅳ collagen protein standard substance, sufficient glycocalicin standard items, neutrophil leucocyte gelatinase correlation rouge
Matter transporter standard items and liver type fatty acid binding protein standard items；
Type Ⅳ collagen protein monoclonal antibody, sufficient glycocalicin monoclonal antibody, neutrophil leucocyte are coated in the gold-labelled pad
Gelatinase associated lipocalin monoclonal antibody and liver type fatty acid binding protein monoclonal antibody；
The first detection reagent item is coated with anti-type Ⅳ collagen protein monoclonal antibody, and the second detection reagent item is coated with
Anti- foot glycocalicin monoclonal antibody, the third detection reagent item are coated with anti-neutrophil leucocyte gelatinase related lipid delivery
Protein monoclonal antibody, the 4th detection reagent article are coated with anti-liver type fatty acid binding protein monoclonal antibody；
The capture reagent strip is coated with the secondary antibody in conjunction with monoclonal antibody coated in the gold-labelled pad.
2. the colloidal gold kit of detection diabetic nephropathy biomarker as described in claim 1, which is characterized in that described
First detection reagent article, the second detection reagent article, third detection reagent article, the 4th detection reagent article and capture reagent strip further include
There is the indicator of the biomarker for quantitative biological sample liquid.
3. the colloidal gold kit of detection diabetic nephropathy biomarker as claimed in claim 2, which is characterized in that described
Indicator is enzyme, prothetic group, fluorescent material, bioluminescence substance or radioactive materials.
4. the colloidal gold kit of detection diabetic nephropathy biomarker as described in claim 1, which is characterized in that described
First detection reagent article, the second detection reagent article, third detection reagent article, the 4th detection reagent article are arranged in the cellulose nitrate
Putting in order for plain film is along the sample chromatography direction in sample pad and parallel from big to small according to biomarker molecular weight
Arranged at equal intervals.
5. the colloidal gold kit of detection diabetic nephropathy biomarker as described in claim 1, which is characterized in that described
Preset length is 1.5~2mm, and the secondary antibody is sheep anti-mouse igg or rabbit anti-mouse igg.
6. the colloidal gold kit of detection diabetic nephropathy biomarker as described in claim 1, which is characterized in that described
Sample pad and gold-labelled pad are all made of glass fibre membrane or polyester film material is made.
7. the colloidal gold kit of detection diabetic nephropathy biomarker as claimed in claim 6, which is characterized in that described
Supporting pad is a kind of rigid plastics bottom plate or PVC support baseboard.
8. the colloidal gold kit of detection diabetic nephropathy biomarker as claimed in claim 7, which is characterized in that described
Water absorption pad is made of the water-absorption fiber or water-absorbing sponge material of water absorbing capacity, for adsorbing and draining the life in the gold-labelled pad
Object sample liquids make chromatography direction of the biological sample liquid by the gold-labelled pad by gold labeling antibody compound along sample liquids
It is adsorbed to corresponding detection reagent position or the position of capture reagent strip on the nitrocellulose filter.
9. a kind of detect diabetic nephropathy biomarker using colloidal gold kit as claimed in any one of claims 1 to 8
Method, which is characterized in that this method comprises the following steps:
Urine is obtained as biological sample liquid from subject；
Biological sample liquid is poured at the load position of gold-labelled pad；
The biomarker of biological sample liquid forms colloidal gold antibody compound in conjunction with the corresponding antibodies in gold-labelled pad；
Colloidal gold antibody compound with sample liquids on nitrocellulose membrane chromatographic flow, by the first detection reagent item, second
Detection reagent article, third detection reagent article, the monoclonal antibody in the 4th detection reagent article combine；
Judge whether the color captured on reagent strip changes；
If the color on capture reagent strip changes, the first detection reagent item, the second detection reagent item, third are detected
The color of reagent strip and the 4th detection reagent article is compared the content for obtaining biomarker with the standard items color of sample pad.
10. as claimed in claim 9 using the method for colloidal gold kit detection diabetic nephropathy biomarker, feature
It is, this method further includes following steps:
If the color on capture reagent strip changes, it is determined that the first detection reagent item of colloidal gold kit, the second inspection
Test agent article, third detection reagent article, the 4th detection reagent article have failed.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810301654.6A CN110346570A (en) | 2018-04-04 | 2018-04-04 | Detect the colloidal gold kit and method of diabetic nephropathy biomarker |
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| CN201810301654.6A CN110346570A (en) | 2018-04-04 | 2018-04-04 | Detect the colloidal gold kit and method of diabetic nephropathy biomarker |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111521831A (en) * | 2020-05-18 | 2020-08-11 | 巴迪泰（广西）生物科技有限公司 | N-terminal brain natriuretic peptide precursor detection kit and preparation method thereof |
| CN113917153A (en) * | 2021-09-22 | 2022-01-11 | 北京松果天目健康管理有限公司 | Application of urine protein marker in preparation of kit for detecting clinical stages of diabetic nephropathy |
-
2018
- 2018-04-04 CN CN201810301654.6A patent/CN110346570A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111521831A (en) * | 2020-05-18 | 2020-08-11 | 巴迪泰（广西）生物科技有限公司 | N-terminal brain natriuretic peptide precursor detection kit and preparation method thereof |
| CN111521831B (en) * | 2020-05-18 | 2023-06-09 | 巴迪泰（广西）生物科技有限公司 | N-terminal brain natriuretic peptide precursor detection kit and preparation method thereof |
| CN113917153A (en) * | 2021-09-22 | 2022-01-11 | 北京松果天目健康管理有限公司 | Application of urine protein marker in preparation of kit for detecting clinical stages of diabetic nephropathy |
| CN113917153B (en) * | 2021-09-22 | 2023-11-24 | 北京松果天目健康管理有限公司 | Application of urine protein marker in preparation of kit for detecting clinical stage of diabetic nephropathy |
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