CN106645763B - The Test paper of total cholesterol - Google Patents
The Test paper of total cholesterol Download PDFInfo
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- CN106645763B CN106645763B CN201710000932.XA CN201710000932A CN106645763B CN 106645763 B CN106645763 B CN 106645763B CN 201710000932 A CN201710000932 A CN 201710000932A CN 106645763 B CN106645763 B CN 106645763B
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- G—PHYSICS
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- 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
- G01N33/92—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving lipids, e.g. cholesterol, lipoproteins, or their receptors
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Abstract
The invention discloses the Test papers of total cholesterol.The detection reagent includes the 4-AA and 0.20~0.30g/L substance that show color of the cholesterol esterase of 6~9KU/L, the cholesterol oxidase of 3~5KU/L, the horseradish peroxidase of 15~20KU/L, the ascorbic acid oxidase of 3~5KU/L, 0.20~0.35g/L.Detection reagent of the invention can reach containing there are many specific proportion of certain enzyme progress and detect quickly and detect accurate purpose.Test paper of the invention includes reaction base, conversion zone, hemofiltration layer and hydrophilic layer, these layers, which are superimposed, constitutes a siphon system, further ensures detection quickly and detection is more accurate.
Description
Technical field
The present invention relates to the technical fields of total cholesterol detection, in particular to the Test paper of total cholesterol.
Background technique
Currently, the method for detecting the content in blood with the presence or absence of certain ingredient or the ingredient includes liquid reagent inspection
It surveys and drying chemical reagent paper detects.Drying chemical reagent paper has the characteristics that easy to use, easy to operate, quick result out.
It, can be by class there are many drying chemical reagent papers according to flow path of the detection liquid sample on drying chemical reagent paper
Type, longitudinally perpendicular Test paper are one kind of drying chemical reagent paper.Longitudinally perpendicular Test paper includes the load that will have reaction reagent
Body is overlapped mutually together, and liquid sample is top-down to flow through each reaction reagent layer, finally shows testing result.Such as beauty
The method that state patent US4816224 uses longitudinally perpendicular Test paper.
The area of longitudinally perpendicular its reaction reagent carrier of Test paper is all smaller, therefore less liquid sample is only needed to use
Amount can be completed to detect.Because of individual difference, during finger tip blood sampling, the sample size of acquisition is very rambunctious, example
Such as, the finger tip blood sampling volume of occidentals is often more than Asian.And reaction reagent carrier has certain saturation to the absorption of liquid
Degree, when the blood sample being added dropwise on test paper is excessive, existing dry chemical Test paper can not handle extra blood sample in time.Such as
Longitudinally perpendicular Test paper in the prior art, the test paper include: that bottom plate adheres to and support conversion zone, are covered in conversion zone
Sample separating layer, fiberglass layer and clathrum.Glass and sample separating layer are vertically flowed through after liquid is added drop-wise to clathrum, are finally reached
It is chemically reacted to conversion zone.If be added blood sample amount it is more, reach hold-up conversion zone cannot reabsorb it is more
When remaining sample, these extra blood samples will overflow checking test paper, pollute environment or touch tester.Extra blood
It can also be gathered in reaction reagent layer and form drop, not only influence the interpretations of the detecting instruments to result such as optics, it can also be serious
Pollution detection instrument.
Patent CN 102128918A uses a kind of lock liquid structure, this structure test paper includes: to adhere to and support on bottom plate
Conversion zone is covered with sample-adding layer relative to the other side of bottom plate in conversion zone, there is an adding mouth in sample-adding layer.Conversion zone is the bottom of along
At least side in plate direction overlaps lock liquid component, and lock liquid component is fixed on bottom plate.In bottom plate relative to having at conversion zone
One for reading the detection window of reaction result.All structures pass through the position between the stronger fixing them of bonding sheet.Work as liquid
It is first vertical to flow into conversion zone and absorbed by conversion zone when body sample is added to sample-adding layer.When conversion zone hold-up, liquid is logical
Cross lateral flow measurement mode be diffused into in conversion zone overlapped lock liquid component.Since the speed that lock liquid component absorbs liquid is less than
Or the absorption liquid velocity equal to conversion zone, so the liquid in conversion zone is when surplus liquid is diffused into lock liquid component
Relevant detection reaction is carried out enough, thus effective dose needed for having no effect on detection reaction.When excessive sample is added,
Extra sample is absorbed by lock liquid component, therefore extra sample not will form drop and overflow except test paper.
Above in the prior art, accuracy in detection is to be improved, detects not quick enough.
Summary of the invention
In view of this, one aspect of the present invention is to provide a kind of detection reagent of total cholesterol, detection reagent detection compared with
To be quick, accuracy in detection is higher.
A kind of detection reagent of total cholesterol, it includes the cholesterol esterases of 6~9KU/L, the cholesterol oxygen of 3~5KU/L
Change the 4- ammonia of enzyme, the horseradish peroxidase of 15~20KU/L, the ascorbic acid oxidase of 3~5KU/L, 0.20~0.35g/L
Base antipyrine and 0.20~0.30g/L substance that show color.
It further, also include the Tris-HCl buffer that 0.15~0.25mmol/L pH is 5.5~6.5.
It further, also include 0.80~1.0g/L trehalose；
It preferably, also include 0.1~0.3g/L BSA；
It preferably, also include 0.15~0.25g/L CHAPS.
Further, the substance that show color be selected from one of TOOS, TODB, DAOS, MOAS, phenols, TMB or two kinds with
On.
Another aspect of the present invention is to provide a kind of Test paper of total cholesterol, and Test paper detection is more quick,
Accuracy in detection is higher.
A kind of Test paper of total cholesterol, including reaction base, be set to it is described reaction one surface of base conversion zone,
It is set to the hemofiltration layer on a surface of conversion zone, and is set to the hydrophilic layer on one surface of hemofiltration layer, the table of the conversion zone
Face is coated with such as above-mentioned detection reagent.
Further, the material of the reaction base is polyvinyl chloride, polycarbonate, polyesteramide or polyester；
Preferably, a double-sided adhesive is equipped between reaction base and the conversion zone.
Further, a double-sided adhesive is equipped between the hydrophilic layer and hemofiltration layer.
Further, the hydrophilic layer is hydrophilic film or grenadine.
Further, the hydrophilic layer is hydrophilic film or grenadine.
Further, the reaction base is equipped with hole, and the double-sided adhesive between the reaction base and conversion zone is equipped with hole.
Detection reagent of the invention, containing there are many certain enzymes to carry out specific proportion, can reach detection quickly and detection compared with
For accurate purpose.Test paper of the invention includes reaction base, conversion zone, hemofiltration layer and hydrophilic layer, these layers are superimposed structure
At a siphon system, further ensures detection quickly and detection is more accurate.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of Test paper of the embodiment of the present invention.
Main element symbol description:
100 Test papers
30 reaction bases
31,41 holes
40,70 double-sided adhesives
50 conversion zones
60 hemofiltration layers
80 hydrophilic layers.
Specific embodiment
Unless otherwise defined, all technologies used herein and scientific term have and the common skill of fields of the present invention
The normally understood identical meaning of art personnel.When there is a conflict, the definition in this specification shall prevail.
Term as used herein:
" by ... preparation " it is synonymous with "comprising".Term "comprising" used herein, " comprising ", " having ", " containing "
Or its any other deformation, it is intended that cover non-exclusionism includes.For example, composition, step, method comprising listed elements,
Product or device are not necessarily limited to those elements, but may include not expressly listed other elements or such composition, step
Suddenly, method, product or the intrinsic element of device.
Conjunction " by ... form " exclude any element that do not point out, step or component.If in claim,
This phrase will make claim closed, so that it is not included the material in addition to the material of those descriptions, but relative
Except customary impurities.When phrase " by ... form " be rather than immediately following theme in the clause that appears in claim main body after
When, only it is limited to element described in the clause；Other elements be not excluded the claim as a whole it
Outside.
Equivalent, concentration or other values or parameter are excellent with range, preferred scope or a series of upper limit preferred values and lower limit
When the Range Representation that choosing value limits, this should be understood as specifically disclosing by any range limit or preferred value and any range
Any pairing of lower limit or preferred value is formed by all ranges, regardless of whether the range separately discloses.For example, when open
When range " 1~5 ", described range should be interpreted as including range " 1~4 ", " 1~3 ", " 1~2 ", " 1~2 and 4~
5 ", " 1~3 and 5 " etc..When numberical range is described herein, unless otherwise stated, otherwise the range is intended to include its end
Value and all integers and score in the range.
" mass parts " refer to the basic measurement unit for indicating the mass ratio relationship of multiple components, and 1 part can indicate arbitrary list
Position quality, can such as be expressed as 1g, may also indicate that 2.689g etc..If we say that the mass parts of component A are a parts, the matter of B component
Measuring part is b parts, then it represents that the quality of component A and the mass ratio a:b of B component.Alternatively, indicating that the quality of component A is aK, B group
The quality divided is bK (K is arbitrary number, indicates multiplying factor).It can not misread, unlike mass fraction, all components
The sum of mass parts be not limited to 100 parts of limitation.
"and/or" is used to indicate that one of illustrated situation or both may to occur, for example, A and/or B includes (A
And B) and (A or B)；
In addition, indefinite article "an" before element of the present invention or component and "one" quantitative requirement to element or component
(i.e. frequency of occurrence) unrestriction.Therefore "one" or "an" should be read as including one or at least one, and odd number
The element or component of form also include plural form, unless the obvious purport of the quantity refers to singular.
The detection reagent 52 of total cholesterol of the invention, it includes the cholesterol esterases of 6~9KU/L, the gallbladder of 3~5KU/L
Sterol oxidizing ferment, the horseradish peroxidase of 15~20KU/L, the ascorbic acid oxidase of 3~5KU/L, 0.20~0.35g/L
4-AA and 0.20~0.30g/L substance that show color.
Specifically, the content of cholesterol esterase can for 6KU/L, 6.1KU/L, 6.5KU/L, 7KU/L, 7.5KU/L,
8.5KU/L or 9KU/L etc.；The content of cholesterol oxidase can be 3KU/L, 3.5KU/L, 4KU/L, 4.5KU/L, 5KU/L etc.；
The content of horseradish peroxidase can for 15KU/L, 15.5KU/L, 16KU/L, 17.5KU/L, 19KU/L, 19.5KU/L or
20KU/L etc.；The content of ascorbic acid oxidase can be 3KU/L, 3.5KU/L, 4KU/L, 4.5KU/L, 5KU/L etc.；4- amino
The content of antipyrine can be 0.20g/L, 0.22g/L, 0.25g/L, 0.28g/L, 0.32g/L, 0.34g/L, 0.35g/L
Deng；The content of substance that show color can be 0.20g/L, 0.22g/L, 0.25g/L, 0.28g/L, 0.29g/L, 0.30g/L etc..
Herein, substance that show color can enumerate one of TOOS, TODB, DAOS, MOAS, phenols, TMB or two kinds with
On.Here, TOOS, that is, N- ethyl-N- (2-Hydroxy-3-sulfopropyl)-3-methylaniline sodium salt, No. CAS is 82692-93-1；
Bis- (4- the sulphur butyl) -3- methylaniline disodium salts of TODB, that is, N, N-, No. CAS is 127544-88-1；DAOS, that is, N- ethyl-N-
(2- hydroxyl -3- sulfopropyl) -3'5- dimethoxyaniline sodium salt, No. CAS is 83777-30-4；TMB, that is, 3,3', 5,5'- tetra-
Methyl biphenyl amine, No. CAS is 54827-17-7；Phenols, that is, phenolic developer.
It, can also be slow comprising Tris-HCl in addition to may include above-mentioned raw materials as above-mentioned detection reagent 52 of the invention
Fliud flushing, with the pH stability of maintenance system.The content of Tris-HCl buffer is preferred with 0.15~0.25mmol/L, such as
0.15mmol/L, 0.18mmol/L, 0.20mmol/L, 0.22mmol/L, 0.24mmol/L or 0.25mmol/L etc., preferably
0.20mmol/L etc..
The pH of Tris-HCl buffer is preferably 5.5~6.5, such as 5.5,5.6,5.8,6,6.2,6.5.
It can also further include 0.80~1.0g/L trehalose as above-mentioned detection reagent 52 of the invention, such as sea
The content of algae sugar can be 0.8g/L, 0.82g/L, 0.85g/L, 0.9g/L, 0.95g/L, 0.98g/L, 1g/L etc..
It can also further include 0.1~0.3g/L BSA, such as its content as above-mentioned detection reagent 52 of the invention
It can be 0.1g/L, 0.15g/L, 0.2g/L, 0.25g/L, 0.3g/L etc..Herein, BSA, that is, bovine serum albumin(BSA) (BSA), again
Claim fifth component, be one of cow's serum albumin, include 583 amino acid residues, molecular weight 66.430kDa waits electricity
Point is 4.7.
It can also further include 0.15~0.25g/LCHAPS as above-mentioned detection reagent 52 of the invention, such as
0.15g/L, 0.16g/L, 0.20g/L, 0.22g/L, 0.24g/L, 0.25g/L etc..Herein, CHAPS is [(3- gallbladder amide groups third
Base) dimethylammonio] -1- propane sulfonic acid salt, No. CAS is 75621-03-3.
The form for reacting base 30 can be film or thin plate, a hole 31 can be set, using as Light-Echo detection
It is used.
The surface of conversion zone 50 is used to drip attached detection reagent, and the form of conversion zone 50 can be film.Conversion zone 50 and anti-
Answer a settable double-sided adhesive 40 between base 30.The area of the double-sided adhesive 40 can be less than reaction base 30, can be greater than conversion zone
50。
A hole 41 can be set in the double-sided adhesive 40, which can be aligned in the hole 31 of reaction base 30, in conversion zone 50
When being covered on reaction base 30 by double-sided adhesive 40.
As Test paper 100 of the invention, the form of hemofiltration layer 60 can be film.The area of hemofiltration layer 60 can be greater than
Conversion zone, its both ends is pasted onto double-sided adhesive 40 when such hemofiltration layer 60 is covered in conversion zone 50.The area of hemofiltration layer 60 can be small
In the size for the double-sided adhesive 40 for bonding it.
A double-sided adhesive 70 can be equipped between hydrophilic layer and hemofiltration layer.The double-sided adhesive can be U-shaped double-sided adhesive and rectangular pair
Face glue, and it is respectively adhered on the upper and lower ends of hemofiltration layer 60.It should be noted that the U-shaped mouth structure of hydrophilic layer 80 and double-sided adhesive 70
The siphon groove built reaches hemofiltration layer.
Above-mentioned hydrophilic layer 80 can be hydrophilic film or grenadine.
It does not address place above and is suitable for the prior art.
Embodiment 1
Step 1: double-sided adhesive 40 sticks in the reaction base 30 in the PET material of certain hardness, it is spare after punching.
Step 2: tearing double-sided adhesive in reaction base 40, reaction film 50 is pasted on the double-sided adhesive 40 of PET sheet, is reacted
Film 50 needs to cover all the hole 31 on pet substrate.
Step 3: preparing detection reagent: the cholesterol oxidation of cholesterol esterase, 3KU/L comprising 6KU/L according to the proportion
Enzyme, the horseradish peroxidase of 15KU/L, the ascorbic acid oxidase of 3KU/L, 0.20g/L 4-AA,
0.20g/L substance that show color, the Tris-HCl buffer of 0.15mmol/L (pH 5.5), 0.80g/L trehalose, 0.1g/L BSA,
0.15g/L CHAPS.Prepared detection reagent is added drop-wise in the hole of reaction diaphragm plate with dispenser, every hole weight is 6mg, drop
The diaphragm plate of complete liquid is put into drying tunnel drying, and drying temperature: 37 DEG C, drying time 20min obtains conversion zone 50.
Step 4: sticking and hemofiltration layer 60 being covered all conversion zone 50.
Step 5: U-shaped double-sided adhesive 70 is sticked around conversion zone 50, using siphon principle, with U-shaped double-sided adhesive 70 and hydrophilic layer
80 (hydrophilic film or grenadines) can fix the amount for inhaling sample as one micro sample absorbing device of composition.
Embodiment 2
In this example detection reagent 52 include the cholesterol esterase of 9KU/L, the cholesterol oxidase of 5KU/L, 20KU/L it is peppery
Root peroxidase, the ascorbic acid oxidase of 5KU/L, the 4-AA of 0.35g/L, 0.30g/L substance that show color,
Tris-HCl buffer, 1.0g/L trehalose, the 0.3g/L BSA, 0.25g/L CHAPS of 0.25mmol/L (pH 5.5),
It is the same as embodiment 1.
Embodiment 3
In this example detection reagent 52 include the cholesterol esterase of 6KU/L, the cholesterol oxidase of 5KU/L, 15KU/L it is peppery
Root peroxidase, the ascorbic acid oxidase of 3KU/L, the 4-AA of 0.20g/L, 0.25g/L substance that show color,
Tris-HCl buffer, 1.0g/L trehalose, the 0.3g/L BSA, 0.25g/L CHAPS of 0.20mmol/L (pH 6), it is other
With embodiment 1.
Embodiment 4
In this example detection reagent 52 include the cholesterol esterase of 9KU/L, the cholesterol oxidase of 3KU/L, 20KU/L it is peppery
Root peroxidase, the ascorbic acid oxidase of 5KU/L, the 4-AA of 0.35g/L, 0.25g/L substance that show color,
Tris-HCl buffer, 0.9g/L trehalose, the 0.2g/L BSA, 0.20g/L CHAPS of 0.20mmol/L (pH 6), it is other
With embodiment 1.
Embodiment 5
Detection reagent 52 includes the cholesterol esterase of 7.5KU/L, the cholesterol oxidase of 4KU/L, 18KU/L in this example
Horseradish peroxidase, the ascorbic acid oxidase of 4KU/L, the 4-AA of 0.27g/L, 0.25g/L substance that show color,
Tris-HCl buffer, 0.9g/L trehalose, the 0.2g/L BSA, 0.20g/L CHAPS of 0.20mmol/L (pH 6), it is other
With embodiment 1.
Comparative example
Certain hospital's test sample.
Evaluation method:
The Test paper 100 of the total cholesterol of embodiment 5 is inserted directly into self-control dry type chemical analysis instrument, after 6 minutes
As a result it is shown on instrument.Table 1 is to make the test result of dry type chemical analysis instrument by oneself with comparative example test result contrast table,
Its correlation reaches 0.997.It can be directly used for clinically testing.
Table 1, hospital's test sample test result compared with total cholesterol test result of the present invention
Test paper 100 of the invention has the advantage that anti-interference ability is stronger, has effectively eliminated red blood cell and interior
The interference of source property substance, and the concentration of the total cholesterol of whole blood can accurately be tested in 6 minutes, test result and tradition
Method compared to test result it is more acurrate.It is used together with company homemade portable dry type chemical analyzer, more meets clinic
The requirement on side is suitble to hospital emergency, ward, community hospital, domestic consumer etc. to use.
The numberical range of each technological parameter as involved in the present invention can not all embody in the above-described embodiments,
As long as but those skilled in the art's envisioned any numerical value fallen into the above-mentioned numberical range completely can be implemented this
Invention also includes any combination of occurrence in several numberical ranges certainly.Herein, due to space considerations, be omitted to
Out in certain one or more numberical range occurrence embodiment, this disclosure for being not to be construed as technical solution of the present invention do not fill
Point.
The Applicant declares that the present invention is explained by the above embodiments detailed process equipment and process flow of the invention,
But the present invention is not limited to the above detailed process equipment and process flow, that is, it is above-mentioned detailed not mean that the present invention must rely on
Process equipment and process flow could be implemented.It should be clear to those skilled in the art, any improvement in the present invention,
Addition, concrete mode selection of equivalence replacement and auxiliary material component ingredient to each raw material of product of the present invention etc., fall in of the invention
In protection scope.
Claims (3)
1. a kind of Test paper of total cholesterol, which is characterized in that including reaction base, be set to one surface of reaction base
Conversion zone, be set to conversion zone a surface hemofiltration layer, and be set to the hydrophilic layer on one surface of hemofiltration layer, it is described anti-
The surface of layer is answered to be coated with the detection reagent of total cholesterol；
The first double-sided adhesive is equipped between reaction base and the conversion zone；The area of first double-sided adhesive is less than the reactive group
Layer；
The second double-sided adhesive is equipped between the hydrophilic layer and hemofiltration layer；Second double-sided adhesive is U-shaped double-sided adhesive；
The area of the hemofiltration layer is greater than the conversion zone, and is less than the area of first double-sided adhesive；
When hemofiltration layer is covered in the conversion zone, both ends are pasted on first double-sided adhesive；
The reaction base is equipped with the first hole, to be used as Light-Echo detection；
First double-sided adhesive between the reaction base and conversion zone is equipped with the second hole；
When the conversion zone is overlying on reaction base by first double faced adhesive tape, second hole and the base of reacting
The first hole alignment；
Wherein, the detection reagent of the total cholesterol includes the cholesterol oxygen of the cholesterol esterase of 6~8.5KU/L, 3~5KU/L
Change the 4- ammonia of enzyme, the horseradish peroxidase of 15~20KU/L, the ascorbic acid oxidase of 3~5KU/L, 0.20~0.35g/L
The Tris-HCl that base antipyrine, 0.20~0.30g/L substance that show color and 0.15~0.25mmol/L pH are 5.5~6.5 is buffered
Liquid；
It also includes 0.80~1.0g/L trehalose；
It also includes 0.1~0.3g/L bovine serum albumin(BSA)；
It also includes 0.15~0.25g/L [(3- gallbladder amidopropyl) dimethylammonio] -1- propane sulfonic acid salt；
The substance that show color is selected from N- ethyl-N- (2-Hydroxy-3-sulfopropyl)-3-methylaniline sodium salt, N, bis- (the 4- sulphur fourths of N-
Base) -3- methylaniline disodium salt, N- ethyl-N- (2- hydroxyl -3- sulfopropyl) -3'5- dimethoxyaniline sodium salt, phenols, 3,
One of 3', 5,5'- tetramethyl benzidine or two kinds or more.
2. Test paper according to claim 1, which is characterized in that the material of the reaction base is polyvinyl chloride, gathers
Carbonic ester, polyesteramide or polyester.
3. Test paper according to claim 2, which is characterized in that the hydrophilic layer is hydrophilic film or grenadine.
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| CN107402209B (en) * | 2017-09-15 | 2020-04-07 | 光景生物科技(苏州)有限公司 | Test strip for detecting low-density lipoprotein cholesterol in serum and preparation method thereof |
| CN108333175B (en) * | 2018-01-18 | 2021-06-29 | 青岛汉唐生物科技有限公司 | Total cholesterol detection 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 |
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| CN110632331A (en) * | 2019-10-31 | 2019-12-31 | 无锡锦帛诚医疗器械科技有限公司 | Dry tablet reagent for quantitative detection of total cholesterol |
| CN110791546A (en) * | 2019-10-31 | 2020-02-14 | 无锡锦帛诚医疗器械科技有限公司 | Dry tablet reagent for quantitative detection of urea nitrogen |
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