CN112881683A - Whole blood antibody detection method for treponema pallidum - Google Patents
Whole blood antibody detection method for treponema pallidum Download PDFInfo
- Publication number
- CN112881683A CN112881683A CN202110037629.3A CN202110037629A CN112881683A CN 112881683 A CN112881683 A CN 112881683A CN 202110037629 A CN202110037629 A CN 202110037629A CN 112881683 A CN112881683 A CN 112881683A
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- Prior art keywords
- blood
- blood sample
- buffer solution
- fingertip
- treponema pallidum
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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
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56911—Bacteria
-
- 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
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
- G01N33/54346—Nanoparticles
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2469/00—Immunoassays for the detection of microorganisms
- G01N2469/20—Detection of antibodies in sample from host which are directed against antigens from microorganisms
Abstract
The invention discloses a whole blood antibody detection method for treponema pallidum, which relates to the technical field of whole blood sample pretreatment, and comprises the following steps: the fingertip blood sampling needle punctures the fingertip; 0.5 ml of washing solution is absorbed by a polyethylene dropper to elute about 100ul of fingertip blood (two drops), and the fingertip blood is dripped into a blood sample collecting tube; taking another polyethylene dropper to suck 0.2 ml of buffer solution and drip the buffer solution into a blood sample collecting tube, and blowing and beating the buffer solution for three times to homogenize the solution; taking another polyethylene dropper to suck 0.2 ml of buffer solution to be dripped into the blood sample collecting pipe, blowing and beating for three times to homogenize, and standing the blood sample collecting pipe for 10 minutes at the room temperature of 10-35 ℃; the pretreated sample was aspirated from the blood sample collection tube with a third polyethylene pipette (0.5 ml). The method improves the detection sensitivity of the syphilis RPR antibody by using a newly designed buffer solution and colloidal gold test paper, thereby realizing earlier treatment and intervention on syphilis patients.
Description
Technical Field
The invention relates to the technical field of whole blood sample pretreatment, in particular to a whole blood antibody detection method for treponema pallidum.
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 the treponema pallidum antibody RPR in a whole blood sample is detected, the detection is usually carried out after four weeks of high-risk exposure, and the syphilis antibody RPR content in the blood is higher, so that the syphilis antibody cannot be detected earlier.
Disclosure of Invention
The invention aims to provide a method for detecting treponema pallidum whole blood antibody.
In order to achieve the purpose, the invention provides the following technical scheme: a method for detecting treponema pallidum whole blood antibody comprises the following steps:
s1, pricking the fingertip by the fingertip blood sampling needle;
s2, sucking 0.5 ml of washing solution by a polyethylene dropper to elute about 100ul of fingertip blood (two drops), and dropping the washing solution into a blood sample collection tube;
s3, taking another polyethylene dropper to suck 0.2 ml of buffer solution to drop into the blood sample collecting tube, and blowing and beating for three times to homogenize;
s4, taking another polyethylene dropper to suck 0.2 ml of buffer solution to drop into the blood sample collecting tube, and blowing and beating for three times to homogenize;
s5, standing the blood sample collection tube for 10 minutes at the room temperature of 10-35 ℃;
s6, 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.
Preferably, the washing solution comprises: sodium acetate, wherein the molar concentration of the sodium acetate is 10mM, and the pH value of the binding buffer is 9.0.
Preferably, the buffer comprises: tris hydrochloride, heparin sodium, NP40 neutral surfactant and Benzonase nuclease, wherein the pH value of the buffer solution is 8.0;
the molar concentration of the Tris hydrochloride is 290-310 mM, the addition amount of the heparin sodium is 9-11 IU, the mass percentage content of the NP40 neutral surfactant is 0.084-0.12%, and the addition amount of the Benzonase nuclease is 0.8-1.2 IU.
Preferably, the molar concentration of the Tris hydrochloride is 300mM, the addition amount of the heparin sodium is 10IU, the mass percentage content of the NP40 neutral surfactant is 0.1%, and the addition amount of the Benzonase nuclease is 1 IU.
Preferably, the colloidal gold test strip is a colloidal gold test strip containing cardiolipin.
Compared with the prior art, the invention has the beneficial effects that:
the method improves the detection sensitivity of the syphilis RPR antibody by using a newly designed buffer solution and colloidal gold test paper, thereby realizing earlier treatment and intervention on syphilis patients.
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 method for detecting treponema pallidum whole blood antibody, which comprises the following steps:
firstly, a fingertip blood sampling needle punctures a fingertip;
then, about 100ul of fingertip blood (two drops) was eluted by sucking 0.5 ml of a washing solution with a polyethylene pipette and dropped into the blood sample collection tube, wherein the washing solution included: sodium acetate, wherein the molar concentration of the sodium acetate is 10mM, and the pH value of the binding buffer is 9.0;
next, another polyethylene pipette was pipetted 0.2 ml of buffer into the blood sample collection tube and pipetted three times to homogenize, the buffer comprising: tris hydrochloride, heparin sodium, NP40 neutral surfactant and Benzonase nuclease, wherein the pH value of the buffer solution is 8.0, the molar concentration of the Tris hydrochloride is 300mM, the addition amount of the heparin sodium is 10IU, the mass percentage content of the NP40 neutral surfactant is 0.1%, and the addition amount of the Benzonase nuclease is 1 IU;
then taking another polyethylene dropper to suck 0.2 ml of buffer solution to be dripped into the blood sample collecting pipe, and blowing and beating for three times to homogenize;
then, standing the blood sample collection tube for 10 minutes at the room temperature of 10-35 ℃;
and finally, sucking 0.5 ml of the pretreated sample from the blood sample collecting pipe by using a third polyethylene dropper, dropping the sample on a sample pad of a colloidal gold test strip, wherein the colloidal gold test strip is the colloidal gold test strip containing the cardiolipin, and standing for 15-20 minutes to read the result.
The method is suitable for the detection of the RPR antibody in blood after the high-risk exposure of the treponema pallidum for 5 days, and the detection sensitivity of the RPR antibody of the syphilis is improved by using the newly designed buffer solution and the colloidal gold test paper, so that the earlier treatment and intervention of syphilis patients are realized.
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 (5)
1. A method for detecting treponema pallidum whole blood antibody is characterized in that: the method comprises the following steps:
s1, pricking the fingertip by the fingertip blood sampling needle;
s2, sucking 0.5 ml of washing solution by a polyethylene dropper to elute about 100ul of fingertip blood (two drops), and dropping the washing solution into a blood sample collection tube;
s3, taking another polyethylene dropper to suck 0.2 ml of buffer solution to drop into the blood sample collecting tube, and blowing and beating for three times to homogenize;
s4, taking another polyethylene dropper to suck 0.2 ml of buffer solution to drop into the blood sample collecting tube, and blowing and beating for three times to homogenize;
s5, standing the blood sample collection tube for 10 minutes at the room temperature of 10-35 ℃;
s6, 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.
2. The method for detecting treponema pallidum whole blood antibody according to claim 1, wherein: the washing liquid comprises: sodium acetate, wherein the molar concentration of the sodium acetate is 10mM, and the pH value of the binding buffer is 9.0.
3. The method for detecting treponema pallidum whole blood antibody according to claim 1, wherein: the buffer solution comprises: tris hydrochloride, heparin sodium, NP40 neutral surfactant and Benzonase nuclease, wherein the pH value of the buffer solution is 8.0;
the molar concentration of the Tris hydrochloride is 290-310 mM, the addition amount of the heparin sodium is 9-11 IU, the mass percentage content of the NP40 neutral surfactant is 0.084-0.12%, and the addition amount of the Benzonase nuclease is 0.8-1.2 IU.
4. The method for detecting treponema pallidum whole blood antibody according to claim 3, wherein: the molar concentration of the Tris hydrochloride is 300mM, the addition amount of the heparin sodium is 10IU, the mass percentage content of the NP40 neutral surfactant is 0.1%, and the addition amount of the Benzonase nuclease is 1 IU.
5. The method for detecting treponema pallidum whole blood antibody according to claim 1, wherein: the colloidal gold test strip is a colloidal gold test strip containing cardiolipin.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110037629.3A CN112881683A (en) | 2021-01-12 | 2021-01-12 | Whole blood antibody detection method for treponema pallidum |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110037629.3A CN112881683A (en) | 2021-01-12 | 2021-01-12 | Whole blood antibody detection method for treponema pallidum |
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| Publication Number | Publication Date |
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| CN112881683A true CN112881683A (en) | 2021-06-01 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN202110037629.3A Pending CN112881683A (en) | 2021-01-12 | 2021-01-12 | Whole blood antibody detection method for treponema pallidum |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4738932A (en) * | 1985-12-03 | 1988-04-19 | Advanced Polymer Systems, Inc. | Reaginic test for syphilis |
| CN101661040A (en) * | 2009-09-10 | 2010-03-03 | 杭州艾力康医药科技有限公司 | Spittle rapid detection strip for treponema pallidum antibody |
| US20140273027A1 (en) * | 2013-03-13 | 2014-09-18 | Awareness Technology Inc. | Serological Methods and Diagnostic Tests for Syphilis Antibodies |
-
2021
- 2021-01-12 CN CN202110037629.3A patent/CN112881683A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4738932A (en) * | 1985-12-03 | 1988-04-19 | Advanced Polymer Systems, Inc. | Reaginic test for syphilis |
| CN101661040A (en) * | 2009-09-10 | 2010-03-03 | 杭州艾力康医药科技有限公司 | Spittle rapid detection strip for treponema pallidum antibody |
| US20140273027A1 (en) * | 2013-03-13 | 2014-09-18 | Awareness Technology Inc. | Serological Methods and Diagnostic Tests for Syphilis Antibodies |
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