CN112881700A - Phosphate buffer solution for whole blood sample pretreatment and use method thereof - Google Patents
Phosphate buffer solution for whole blood sample pretreatment and use method thereof Download PDFInfo
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- CN112881700A CN112881700A CN202110037628.9A CN202110037628A CN112881700A CN 112881700 A CN112881700 A CN 112881700A CN 202110037628 A CN202110037628 A CN 202110037628A CN 112881700 A CN112881700 A CN 112881700A
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- phosphate buffer
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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
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/585—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with a particulate label, e.g. coloured latex
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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
- G01N33/5306—Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
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Abstract
The invention discloses a phosphate buffer solution for whole blood sample pretreatment and a use method thereof, relating to the technical field of whole blood sample pretreatment, wherein the phosphate buffer solution comprises disodium hydrogen phosphate, sodium dihydrogen phosphate, a CA630 surfactant and Benzonase nuclease, and the pH value of the phosphate buffer solution is 7.4; the molar concentration of the disodium hydrogen phosphate is 4-6 mM, the molar concentration of the sodium dihydrogen phosphate is 4-6 mM, the mass percentage content of the CA630 surfactant is 0.08-0.13%, and the addition amount of the Benzonase nuclease is 0.8-1.3 IU. According to the invention, the whole blood sample is pretreated by using a phosphate buffer solution with a brand-new design, so that the blood sample to be detected can be smoothly migrated and detected on the colloidal gold.
Description
Technical Field
The invention relates to the technical field of whole blood sample pretreatment, in particular to a phosphate buffer solution for whole blood sample pretreatment and a use method thereof.
Background
Colloidal gold is called colloidal gold, which is obtained by polymerizing chloroauric acid (HAuCl4) into gold particles with a certain size under the action of a reducing agent such as white phosphorus, ascorbic acid, sodium citrate, tannic acid and the like, forming a negatively charged hydrophobic colloidal solution due to electrostatic interaction and forming a stable colloidal state due to electrostatic interaction. The colloidal gold has negative charge in weak alkali environment, can form firm combination with the positive charge group of protein molecules, and does not influence the biological characteristics of the protein because the combination is electrostatic combination.
Colloidal gold, in addition to binding to proteins, can also bind to many other biomacromolecules, such as SPA, PHA, ConA, and the like. According to some physical properties of colloidal gold, such as high electron density, particle size, shape and color reaction, and immune and biological characteristics of the conjugate, colloidal gold is widely used in the fields of immunology, histology, pathology and cell biology.
When a whole blood sample is used as a sample to be detected of the colloidal gold test strip, the migration of the sample to be detected on the colloidal gold test strip is influenced because blood components are complex, protein content is high, cells are various, and various substances are easy to crosslink and/or interact. Meanwhile, protein or nucleic acid components in blood are easy to combine and shield antigens or antibodies fixed on a colloidal gold test paper detection line. The above two factors affect the reading of the detection result.
Disclosure of Invention
The invention aims to provide a phosphate buffer solution for whole blood sample pretreatment and a use method thereof.
In order to achieve the purpose, the invention provides the following technical scheme: a phosphate buffer for whole blood sample pretreatment, the phosphate buffer comprising disodium hydrogen phosphate, sodium dihydrogen phosphate, CA630 surfactant, Benzonase nuclease, the pH of the phosphate buffer being 7.4;
the molar concentration of the disodium hydrogen phosphate is 4-6 mM, the molar concentration of the sodium dihydrogen phosphate is 4-6 mM, the mass percentage content of the CA630 surfactant is 0.08-0.13%, and the addition amount of the Benzonase nuclease is 0.8-1.3 IU.
Preferably, the molar concentration of the disodium hydrogen phosphate is 5mM, the molar concentration of the sodium dihydrogen phosphate is 5mM, the mass percentage content of the CA630 surfactant is 0.1%, and the addition amount of the Benzonase nuclease is 1 IU.
A method of using a phosphate buffer for whole blood sample pretreatment comprising the steps of:
s1, pricking the fingertip by the fingertip blood sampling needle;
s2, sucking 0.5 ml of 150mM NaCl by using a polyethylene dropper to elute 100ul of fingertip blood, and dripping the fingertip blood into a blood sample collecting tube;
s3, taking another polyethylene dropper to suck 0.2 ml of phosphate buffer solution and drip the phosphate buffer solution into the blood sample collecting tube, and blowing and beating the phosphate buffer solution for three times to homogenize the mixture;
s4, standing the blood sample collection tube for 10 minutes in an environment of 10-35 ℃;
s5, sucking 0.5 ml of the pretreated sample from the blood sample collecting tube by using a third polyethylene dropper, dropping the pretreated sample on a sample pad of a colloidal gold test strip, and standing for 15-20 minutes to read the result.
Compared with the prior art, the invention has the beneficial effects that:
the product pretreats the whole blood sample by using a phosphate buffer solution with brand-new design, so that the blood sample to be detected can be smoothly migrated and detected on the colloidal gold.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments.
The embodiment provides a phosphate buffer solution for whole blood sample pretreatment, wherein the phosphate buffer solution comprises disodium hydrogen phosphate, sodium dihydrogen phosphate, a CA630 surfactant and Benzonase nuclease, and the pH value of the phosphate buffer solution is 7.4;
specifically, the molar concentration of the disodium hydrogen phosphate is 5mM, the molar concentration of the sodium dihydrogen phosphate is 5mM, the mass percentage content of the CA630 surfactant is 0.1%, and the addition amount of the Benzonase nuclease is 1 IU.
The prepared phosphate buffer solution is suitable for the situation that the antigen or the antibody to be detected is free in blood or is not tightly combined with cell membranes.
After the phosphate buffer was dispensed, the following products were then prepared:
a fingertip blood sampling needle;
150mM NaCl sterile saline;
a disposable polyethylene dropper;
a blood sample collection tube.
After the preparation is complete, the method can be used by the following method:
firstly, sampling blood from a fingertip, and puncturing the fingertip by using a needle;
then, 0.5 ml of 150mM NaCl is absorbed by a polyethylene dropper to elute 100ul of fingertip blood, and the fingertip blood is dripped into a blood sample collecting tube;
then another polyethylene dropper is taken to suck 0.2 ml of phosphate buffer solution and is dripped into the blood sample collecting pipe, and the mixture is blown and beaten for three times to be homogenized;
then, the blood sample collection tube is kept still for 10 minutes in the environment of 10-35 ℃;
then, a third polyethylene dropper is used for sucking 0.5 ml of the pretreated sample from the blood sample collecting tube, and the pretreated sample is dripped on a sample pad of the colloidal gold test strip and stands for 15 to 20 minutes, so that the result can be read.
The product pretreats the whole blood sample by using a phosphate buffer solution with brand-new design, so that the blood sample to be detected can be smoothly migrated and detected on the colloidal gold.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Claims (3)
1. A phosphate buffer for use in the pretreatment of a whole blood sample, characterized by: the phosphate buffer solution comprises disodium hydrogen phosphate, sodium dihydrogen phosphate, a CA630 surfactant and Benzonase nuclease, and the pH value of the phosphate buffer solution is 7.4;
the molar concentration of the disodium hydrogen phosphate is 4-6 mM, the molar concentration of the sodium dihydrogen phosphate is 4-6 mM, the mass percentage content of the CA630 surfactant is 0.08-0.13%, and the addition amount of the Benzonase nuclease is 0.8-1.3 IU.
2. The phosphate buffer solution for the pretreatment of a whole blood sample according to claim 1, wherein: the molar concentration of the disodium hydrogen phosphate is 5mM, the molar concentration of the sodium dihydrogen phosphate is 5mM, the mass percentage content of the CA630 surfactant is 0.1%, and the addition amount of the Benzonase nuclease is 1 IU.
3. A method of using a phosphate buffer for the pretreatment of a whole blood sample, comprising the steps of:
s1, pricking the fingertip by the fingertip blood sampling needle;
s2, sucking 0.5 ml of 150mM NaCl by using a polyethylene dropper to elute 100ul of fingertip blood, and dripping the fingertip blood into a blood sample collecting tube;
s3, taking another polyethylene dropper to suck 0.2 ml of phosphate buffer solution and drip the phosphate buffer solution into the blood sample collecting tube, and blowing and beating the phosphate buffer solution for three times to homogenize the mixture;
s4, standing the blood sample collection tube for 10 minutes in an environment of 10-35 ℃;
s5, sucking 0.5 ml of the pretreated sample from the blood sample collecting tube by using a third polyethylene dropper, dropping the pretreated sample on a sample pad of a colloidal gold test strip, and standing for 15-20 minutes to read the result.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110037628.9A CN112881700A (en) | 2021-01-12 | 2021-01-12 | Phosphate buffer solution for whole blood sample pretreatment and use method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110037628.9A CN112881700A (en) | 2021-01-12 | 2021-01-12 | Phosphate buffer solution for whole blood sample pretreatment and use method thereof |
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| CN112881700A true CN112881700A (en) | 2021-06-01 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023059921A3 (en) * | 2021-10-07 | 2023-05-19 | Intelligent Optical Systems, Inc. | Enhanced lateral flow assays and devices for detecting analytes in blood samples |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080160528A1 (en) * | 2005-03-02 | 2008-07-03 | Lorenz Michael G | Use of Nucleases for Degrading Nucleic Acids in the Presence of Chaotropic Agents and/or Surfactants |
| WO2011132089A2 (en) * | 2010-02-26 | 2011-10-27 | Dalhousie University | Methods for tissue decellularization |
| CN108139402A (en) * | 2015-09-04 | 2018-06-08 | 加利福尼亚大学董事会 | The method and apparatus for collecting for the analyte of clinical practice, extracting, concentrate and detecting |
| CN111549101A (en) * | 2020-06-09 | 2020-08-18 | 无锡市申瑞生物制品有限公司 | Preservation solution for biological sample nucleic acid detection and application |
-
2021
- 2021-01-12 CN CN202110037628.9A patent/CN112881700A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080160528A1 (en) * | 2005-03-02 | 2008-07-03 | Lorenz Michael G | Use of Nucleases for Degrading Nucleic Acids in the Presence of Chaotropic Agents and/or Surfactants |
| WO2011132089A2 (en) * | 2010-02-26 | 2011-10-27 | Dalhousie University | Methods for tissue decellularization |
| CN108139402A (en) * | 2015-09-04 | 2018-06-08 | 加利福尼亚大学董事会 | The method and apparatus for collecting for the analyte of clinical practice, extracting, concentrate and detecting |
| CN111549101A (en) * | 2020-06-09 | 2020-08-18 | 无锡市申瑞生物制品有限公司 | Preservation solution for biological sample nucleic acid detection and application |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023059921A3 (en) * | 2021-10-07 | 2023-05-19 | Intelligent Optical Systems, Inc. | Enhanced lateral flow assays and devices for detecting analytes in blood samples |
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