CN106442079A - Hemofiltration sample pad and preparation method thereof - Google Patents
Hemofiltration sample pad and preparation method thereof Download PDFInfo
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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Abstract
The invention relates to a hemofiltration sample pad. According to a preparation mode, a certain proportion of human erythrocyte antibodies, trihydroxymethyl aminomethane, casein, polyvinylpyrrolidone, surfactants and the like are prepared into a treatment solution according to different product properties in accordance with different sequences; the prepared solution is subjected to pH regulation so as to form a final treatment solution; the prepared treatment solution is uniformly processed on glass fiber and is put into a 37-DEG C drying box to be dried for 8 to 24 hours. The hemofiltration sample pad prepared by the method has the advantages that the whole blood sample is separated under the condition of not influencing the sample running board; a fast diagnosis reagent subjected to sample feeding has clean background without phenomena such as blood residue traces, flooding, line whitening or rail lines.
Description
Technical field
The present invention relates to the whole blood seperation film in pharmaceutical test field, more particularly, to Whole blood assay and preparation method thereof.
Technical background
Hemofiltration sample pad uses main material at this stage is whole blood seperation film, is also called hemofiltration film or plasma separation membrane, can
Blood plasma in quick separating whole blood and no haemolysis, allow the terminal of product the use of client need not be serum centrifugal blood, and
Further saving detection time increases efficiency, directly uses peripheral blood or new blood.Hemofiltration UF membrane is complete
Blood retains red blood cell in whole blood, leucocyte by physical separation mode.
Add whole blood seperation film on testing reagent bar, without extra means, be convenient for carrying and use；The speed of separated plasma
Degree is fast；Select suitable hemofiltration film, haemolysis possibility very little.But, there is certain defect, such as in existing hemofiltration film：1st, it is used in inspection
When testing reagent strip detection whole blood sample, often can affect to detect the race plate speed of sample and testing result, occur blood residuals vestige,
The phenomenons such as lines whiting.2nd, prior art is directed to the difference of final products packaged form, different to the length requirement of film, need into
Row is targetedly debugged；Need when the 3rd, using hemofiltration film to be further added by one layer of sealer adhesive tape so that reagent strip thickness itself increases, no
Beneficial to production operation；4th, during viscous hemofiltration film, should be noted that positive and negative operates, in a large amount of productions, operation easily occurs
Error.
Content of the invention
The technical problem to be solved be provide that a kind of operation is simpler, the more preferable sample pad of hemofiltration effect and its
Preparation method.
A kind of hemofiltration sample pad, including the processing solution of glass fibre and glass fibre, described processing solution includes resisting
RBC, surfactant, phosphate buffer and purified water, described processing solution also includes trishydroxymethylaminomethane, casein
And polyvinylpyrrolidone.
The preparation method of the described processing solution of described hemofiltration sample pad is as follows：
1) take the purified water of processing solution aequum 0.9, add three hydroxyls of 0.04～0.06M/L in order in purified water
Aminomethane, the casein of 0.15～0.25mM, the polyvinylpyrrolidone of 0.15～0.35mM, the group of previous addition
Point stirring adds next component after being completely dissolved, and stirs to being completely dissolved；
2) be sequentially added in the solution prepared the sodium carbonate of 0.005～0.015M, required processing solution 0.6～
The anti-RBC of mouse of 1.1% surfactant and required processing solution 3～5%, the component stirring of previous addition is completely molten
Add next component after solution, stir to being completely dissolved；
3) solution ph is adjusted to 8.0 ± 0.1；
4) with purified water, prepared solution is settled to required liquor capacity, and repetition measurement pH value is 8.0 ± 0.1.
The another preparation method of the described processing solution of described hemofiltration sample pad is as follows：
1) take the purified water of processing solution aequum 0.9, add the three hydroxyl first of 0.04～0.06M in order in purified water
Base aminomethane, the casein of 0.15～0.25mM, the polyvinylpyrrolidone of 0.15～0.35mM, previous component stirring is completely
Add next component after dissolving, stir to being completely dissolved；
2) in step 1) prepare solution in be sequentially added into：The Na of 0.01～0.015M2CO3, required processing solution 0.2～
0.6% surfactant, the stirring of previous component adds next component after being completely dissolved, and stirs to being completely dissolved；
3) solution ph is adjusted to 8.0 ± 0.1；
4) add in the solution adjust pH value：The anti-RBC of mouse of required processing solution 3～5%, stirs to solution
Clarification；
5) with purified water, solution is settled to required processing solution volume, and repetition measurement pH value is 8.0 ± 0.1.
The another preparation method of the described processing solution of described hemofiltration sample pad is as follows：
1) take the purified water of processing solution aequum 0.7, add the three hydroxyl first of 0.04～0.06M in order in purified water
Base aminomethane, the polyvinylpyrrolidone of 0.1～0.3mM, 0.02～0.1% surfactant, 0.15～0.25mM
Casein, the ethylenediamine tetra-acetic acid of 0.02～0.04M, the protein stabiliser of required processing solution 8～12%, required process are molten
The anti-RBC of sheep source property of liquid 2～5%, the stirring of previous component adds next component after being completely dissolved, and stirs to being completely dissolved；
2) solution ph is adjusted to 8.0 ± 0.1；
3) pH value will be adjusted obtain solution purified water and be settled to required liquor capacity, and repetition measurement pH value will be 8.0 ± 0.1；
The another preparation method of the described processing solution of described hemofiltration sample pad is as follows：
1) take the purified water of processing solution aequum 0.8, purified water is sequentially added into the three hydroxyl first of 0.08～0.12M
Base aminomethane, the polyvinylpyrrolidone of 0.3～0.5mM, the ethylenediamine tetra-acetic acid of 0.02～0.04M, required processing solution 8
～12% protein stabiliser, the surfactant of required processing solution 0.2～0.6%, the casein of 0.15～0.25mM, front
One component stirring adds next component after being completely dissolved, and stirs to being completely dissolved；
2) solution ph is adjusted to 8.0 ± 0.1；
3) add in the solution adjust pH value：The anti-RBC of mouse of required processing solution 3～5%, stirs to solution
Clarification；
4) solution is settled to required processing solution volume, and repetition measurement pH value is 8.0 ± 0.1.
Preferably, described adjustment pH value solution be hydrochloric acid or sodium hydroxide solution any one.
The preparation method of described hemofiltration sample pad, comprises the following steps：
1) prepare described processing solution；
2) by the processing solution preparing uniform treatment on the glass fibers；
3) glass fibre processing treated solution is placed in 37 DEG C of environment and dries 8～24 hours.
Beneficial effects of the present invention：
1st, the component proportion of the processing solution of this hemofiltration sample pad and processing method can make this hemofiltration sample pad in not shadow
Ring in the case that sample runs plate and whole blood sample is carried out separating well, abundant experimental results and evidence obtaining show using this hemofiltration
Fast diagnosis reagent bar clean background after sample pad sample-adding no blood residuals vestige, no Flooding, no lines whiting, path
Etc. phenomenon.
2nd, the compound method of the processing solution of sample pad makes this hemofiltration sample pad can improve the homogeneity of sample, reconciles mark
This rate of release, improves solvability in sample pad for the tested substance, eliminates false positive to a certain extent, makes in detection line
Immune protein conjugate more stable.
3rd, this hemofiltration sample pad will not be adjusted with the change of reagent strip width；Mould adhesive tape need not be sealed, simplify production work
Skill；Production operation convenient purification need not special differentiation sample pad positive and negative, improve production efficiency；In the case that product quality is constant,
Reduce production cost, promote enterprise development.
Specific embodiment
This hemofiltration sample pad is the fiberglass packing that a kind of buffered salting liquid was processed.The process of this hemofiltration sample pad is molten
Liquid comprises a certain proportion of human red cell antibody (following shorthand：Anti- RBC), trishydroxymethylaminomethane (following shorthand：
Tris), casein (following shorthand：Casein), polyvinylpyrrolidone (following shorthand：PVP), surfactant, ethylenediamine
Tetraacethyl, protein stabiliser.Add different anti-RBC according to different properties of product and other auxiliary solutes have reached optimal inspection
Survey effect.
The preparation process of this sample pad is as follows：
1st, each component raw material is configured to according to corresponding ratio and order by solution according to different product performance, to preparing
Solution be adjusted pH value to form the optimized buffer system of suitable antigen-antibody reaction；
2nd, solution aequum is calculated according to corresponding glass fibre imbibition coefficient, by the processing solution preparing uniform treatment
On the glass fibers；
3rd, the glass fibre that treated solution was processed is placed in 37 DEG C of baking ovens or drying room and dries 8～24 hours.
Embodiment 1：The hemofiltration sample of human immunodeficiency virus (HIV 1/2) antibody test reagent (emulsion technique) product
Pad is processed
The preparation process of this hemofiltration sample pad is as follows：
1) purified water of prep solution aequum 0.9, is sequentially added into Tris, the 0.15～0.25mM of 0.04～0.06M
Casein, the PVP of 0.15～0.35mM, the stirring of previous component adds next component after being completely dissolved, and stirs to being completely dissolved；
2) in step 1) middle addition in order in the solution prepared：The sodium carbonate of 0.01～0.015M is (hereinafter abbreviated as：
Na2CO3), the surfactant of required processing solution 0.6～1.1%, the anti-RBC of mouse of required processing solution 3～5%, front
One component stirring adds next component after being completely dissolved, and stirs to being completely dissolved, solution is clarified；
3) adjusting pH value with hydrochloric acid or sodium hydroxide solution is 8.0 ± 0.1；
4) with purified water, prepared solution is settled to required liquor capacity, and repetition measurement pH value is 8.0 ± 0.1；
5) process uniform for the processing solution preparing on the glass fibers, and dried in 37 DEG C of baking ovens or drying room
8～24 hours.
Embodiment 2:Hepatitis b virus s antigen detection reagent (colloidal gold method) outturn sample pad is processed
The preparation process of this sample pad is as follows：
1) take the purified water of processing solution aequum 0.7, add the Tris of 0.04～0.06M in order in purified water,
The PVP of 0.1～0.3mM, the surfactant of required processing solution 0.02～0.1%, the Casein of 0.15～0.25mM, 0.02
The ethylenediamine tetra-acetic acid of～0.04M, the protein stabiliser of required processing solution 8～12%, the sheep of required processing solution 2～5%
The anti-RBC of source property, previous component stirring adds next component after being completely dissolved, and stirs to being completely dissolved, solution is clarified；
2) adjusting solution ph with hydrochloric acid or sodium hydroxide solution is 8.0 ± 0.1；
3) solution purified water is settled to required liquor capacity, and repetition measurement pH value is 8.0 ± 0.1；
4) process uniform for the processing solution preparing on the glass fibers, and dried in 37 DEG C of baking ovens or drying room
Dry 8～24 hours.
Embodiment 3:Syphilis helicoid antibody detection reagent (emulsion technique) outturn sample pad is processed
The preparation process of this sample pad is as follows：
1) take the purified water of processing solution aequum 0.9, in order in purified water add 0.04～0.06M Tris,
The Casein of 0.15～0.25mM, the PVP of 0.15～0.35mM, previous component stirring adds next component after being completely dissolved, and stirs
Mix to being completely dissolved, solution is clarified；
2) in step 1) prepare solution in be sequentially added into：The Na of 0.01～0.015M2CO3, required processing solution 0.2～
0.6% surfactant, previous component stirring adds next component after being completely dissolved, and stirs to being completely dissolved, solution is clarified；
3) with hydrochloric acid or NaOH, solution ph is adjusted to 8.0 ± 0.1；
4) add in the solution adjust pH value：The anti-RBC of mouse of required processing solution 3～5%, stirs to solution
Clarification；
5) with purified water, solution is settled to required processing solution volume, and repetition measurement pH value is 8.0 ± 0.1；
6) process uniform for the processing solution preparing on the glass fibers, and dried in 37 DEG C of baking ovens or drying room
8～24 hours.
Embodiment 4:Helicobacter pylori IgG antibody detection reagent (emulsion technique) product hemofiltration sample pad is processed
The preparation process of this sample pad is as follows：
1) take the purified water of processing solution aequum 0.8, by following orders add 0.08～0.12M Tris, 0.3～
The PVP of 0.5mM, the ethylenediamine tetra-acetic acid of 0.02～0.04M, the protein stabiliser of required processing solution 8～12%, required process
Solution 0.2～0.6% surfactant, the Casein of 0.15～0.25mM, previous component stirring adds next after being completely dissolved
Component, stirs to being completely dissolved, solution is clarified；
2) with hydrochloric acid or NaOH, solution ph is adjusted to 8.0 ± 0.1；
3) needed for adding in the solution adjust pH value, the anti-RBC of mouse of processing solution 3～5%, stirs to solution
Clarification；
4) solution is settled to required processing solution volume, and repetition measurement pH is 8.0 ± 0.1；
5) process uniform for the processing solution preparing on the glass fibers, and dried in 37 DEG C of baking ovens or drying room
8～24 hours.
In sum, this sample pad selects different biogenic for the anti-RBC that different product performance is adopted.As the mankind
HIV (HIV 1/2) antibody test reagent (emulsion technique) selects the anti-RBC of mouse, hepatitis b virus s antigen
Detection reagent (colloidal gold method) selects the anti-RBC of sheep source property.
The use of surfactant has cleaning function, can dissolve label, strengthens protein water solubility, eliminates non-spy
The opposite sex combines (covering some miscellaneous sites), is also equipped with certain carrier function simultaneously；PVP can eliminate non-specific adsorption, with
When there is certain Action of Surfactant；The protein materials such as Casein can be with specific adsorption foreign protein (non-specific egg
In vain), eliminate false sun or the false negative in detection process；PH value is adjusted to 8.0 ± 0.1 and can eliminate false positive, sometimes may be used
To change protein conformation, promote more binding sites to expose outside and lead to carrying of sensitivity thus being easier to combine with label
High.
The processing solution for preparing is uniform to be processed on the glass fibers, and carry out drying 8 in 37 DEG C of baking ovens or drying room～
24 hours, preferably attachment on the glass fibers, did not affect this performance of having of hemofiltration sample pad simultaneously can to make solution.
The preparation of fast diagnosis reagent bar and testing result：
Fast diagnosis reagent bar includes hemofiltration sample pad, label pad, NC film and adsorptive pads, and wherein hemofiltration sample pad is tested
This load position of test sample, can be filtered to detection sample and be buffered, be reduced sample ionic strength or acid-base value to inspection
Survey the interference adding；Have by fixing recombinant antigen label be dried in label pad, the antibody in traceable sample therewith
Reaction forms antibody-recombinant antigen label, makes the antibody in sample carry color；Detection line and Quality Control are coated in advance on NC film
Line, can occur immune response combine with antibody-recombinant antigen label, in conjunction with after label in detection line and Quality Control
Assemble at line, interpretation is carried out to result according to the colour developing situation of detection line；Adsorptive pads can by water sorption be liquid sample to
Upper flowing drives the antibody-recombinant antigen label of the formation in label pad to move up, thus the restructuring being gone out with detection line is resisted
Former react.
Hemofiltration sample pad, label pad, NC film and adsorptive pads 4 part are assembled in order, using this hemofiltration sample pad formula
, in the case of keeping excellent checking function (sensitivity, specificity etc.), race plate clean background is without exception for product afterwards,
Save production cost to a great extent, improve operating efficiency.
Fast diagnosis reagent bar using this hemofiltration sample pad formula and preparation technology output passes through many experiments and evidence obtaining
Obtain following result：
Above-mentioned as shown by data uses the fast diagnosis reagent bar clean background no blood residuals after this hemofiltration sample pad sample-adding
Vestige, no Flooding, the no phenomenon such as lines whiting, path.
Described above is exemplary rather than restricted.By described above skilled person realizes that this
The many kind of invention changes and deforms, and it also will fall within the spirit and scope of the invention.
Claims (7)
1. a kind of hemofiltration sample pad, including the processing solution of glass fibre and glass fibre, described processing solution include anti-RBC,
Surfactant, phosphate buffer and purified water are it is characterised in that described processing solution also includes trihydroxy methyl amino first
Alkane, casein and polyvinylpyrrolidone.
2. the preparation method of hemofiltration sample pad according to claim 1 is it is characterised in that the preparation side of described processing solution
Method is as follows：
1) take the purified water of processing solution aequum 0.9, add the trihydroxy methyl ammonia of 0.04～0.06M in order in purified water
Methylmethane, the casein of 0.15～0.25mM, the polyvinylpyrrolidone of 0.15～0.35mM, the component stirring of previous addition
Add next component after being completely dissolved, stir to being completely dissolved；
2) it is sequentially added into the sodium carbonate of 0.005～0.015M, required processing solution 0.6～1.1% in the solution prepared
The anti-RBC of mouse of surfactant and required processing solution 3～5%, the component stirring of previous addition adds after being completely dissolved
Enter next component, stir to being completely dissolved；
3) solution ph is adjusted to 8.0 ± 0.1；
4) with purified water, prepared solution is settled to required liquor capacity, and repetition measurement pH value is 8.0 ± 0.1.
3. the preparation method of hemofiltration sample pad according to claim 1 is it is characterised in that the preparation side of described processing solution
Method is as follows：
1) take the purified water of processing solution aequum 0.9, add the trihydroxy methyl ammonia of 0.04～0.06M in order in purified water
Methylmethane, the casein of 0.15～0.25mM, the polyvinylpyrrolidone of 0.15～0.35mM, previous component stirring is completely dissolved
After add next component, stir to being completely dissolved；
2) in step 1) prepare solution in be sequentially added into：The Na of 0.01～0.015M2CO3, required processing solution 0.2～0.6%
Surfactant, the stirring of previous component adds next component after being completely dissolved, and stirs to being completely dissolved；
3) solution ph is adjusted to 8.0 ± 0.1；
4) add in the solution adjust pH value：The anti-RBC of mouse of required processing solution 3～5%, stirs clear to solution
Clearly；
5) with purified water, solution is settled to required processing solution volume, and repetition measurement pH value is 8.0 ± 0.1.
4. the preparation method of hemofiltration sample pad according to claim 1 is it is characterised in that the preparation side of described processing solution
Method is as follows：
1) take the purified water of processing solution aequum 0.7, add the trihydroxy methyl ammonia of 0.04～0.06M in order in purified water
Methylmethane, the polyvinylpyrrolidone of 0.1～0.3mM, 0.02～0.1% surfactant, the junket egg of 0.15～0.25mM
In vain, the ethylenediamine tetra-acetic acid of 0.02～0.04M, the protein stabiliser of required processing solution 8～12%, required processing solution 2～
The 5% anti-RBC of sheep source property, the stirring of previous component adds next component after being completely dissolved, and stirs to being completely dissolved；
2) solution ph is adjusted to 8.0 ± 0.1；
3) pH value will be adjusted obtain solution purified water and be settled to required liquor capacity, and repetition measurement pH value will be 8.0 ± 0.1.
5. the preparation method of hemofiltration sample pad according to claim 1 is it is characterised in that the preparation side of described processing solution
Method is as follows：
1) take the purified water of processing solution aequum 0.8, purified water is sequentially added into the trihydroxy methyl ammonia of 0.08～0.12M
Methylmethane, the polyvinylpyrrolidone of 0.3～0.5mM, the ethylenediamine tetra-acetic acid of 0.02～0.04M, required processing solution 8～
12% protein stabiliser, the surfactant of required processing solution 0.2～0.6%, the casein of 0.15～0.25mM, previous
Component stirring adds next component after being completely dissolved, and stirs to being completely dissolved；
2) solution ph is adjusted to 8.0 ± 0.1；
3) add in the solution adjust pH value：The anti-RBC of mouse of required processing solution 3～5%, stirs clear to solution
Clearly；
4) solution is settled to required processing solution volume, and repetition measurement pH value is 8.0 ± 0.1.
6. the preparation method of the hemofiltration sample pad according to Claims 2 or 3 or 4 or 5 is it is characterised in that described adjustment pH
Value solution be hydrochloric acid or sodium hydroxide solution any one.
7. the preparation method of the hemofiltration sample pad according to claim 1 or 2 or 3 or 4 or 5 is it is characterised in that include following
Step：
1) prepare described processing solution；
2) by the processing solution preparing uniform treatment on the glass fibers；
3) glass fibre processing treated solution is placed in 37 DEG C of environment and dries 8～24 hours.
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Cited By (1)
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| CN106950373A (en) * | 2017-03-29 | 2017-07-14 | 杭州博拓生物科技股份有限公司 | A kind of half-quantitative detection kits of CA15 3 and preparation technology |
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| CN106442079B (en) | 2020-04-14 |
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