CN107192823A - A kind of B races streptozyme linked immune assay kit - Google Patents
A kind of B races streptozyme linked immune assay kit Download PDFInfo
- Publication number
- CN107192823A CN107192823A CN201710427971.8A CN201710427971A CN107192823A CN 107192823 A CN107192823 A CN 107192823A CN 201710427971 A CN201710427971 A CN 201710427971A CN 107192823 A CN107192823 A CN 107192823A
- Authority
- CN
- China
- Prior art keywords
- races
- streptozyme
- assay kit
- immune assay
- linked immune
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000003149 assay kit Methods 0.000 title claims abstract description 21
- 238000004140 cleaning Methods 0.000 claims abstract description 32
- 238000001514 detection method Methods 0.000 claims abstract description 30
- 239000000758 substrate Substances 0.000 claims abstract description 18
- 238000000034 method Methods 0.000 claims abstract description 14
- 239000000427 antigen Substances 0.000 claims abstract description 11
- 102000036639 antigens Human genes 0.000 claims abstract description 11
- 108091007433 antigens Proteins 0.000 claims abstract description 11
- 239000011324 bead Substances 0.000 claims abstract description 11
- 230000027455 binding Effects 0.000 claims abstract description 8
- 238000009739 binding Methods 0.000 claims abstract description 8
- 239000002184 metal Substances 0.000 claims abstract description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 28
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 claims description 16
- 239000007788 liquid Substances 0.000 claims description 16
- 239000011780 sodium chloride Substances 0.000 claims description 14
- 239000012154 double-distilled water Substances 0.000 claims description 13
- 239000002904 solvent Substances 0.000 claims description 13
- 229960002685 biotin Drugs 0.000 claims description 8
- 235000020958 biotin Nutrition 0.000 claims description 8
- 239000011616 biotin Substances 0.000 claims description 8
- 238000002372 labelling Methods 0.000 claims description 7
- 239000012530 fluid Substances 0.000 claims description 6
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 claims description 5
- 229940005654 nitrite ion Drugs 0.000 claims description 5
- 230000010354 integration Effects 0.000 abstract description 2
- 241000194017 Streptococcus Species 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 108010004729 Phycoerythrin Proteins 0.000 description 2
- 108010090804 Streptavidin Proteins 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000001717 pathogenic effect Effects 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 241000193990 Streptococcus sp. 'group B' Species 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 230000001143 conditioned effect Effects 0.000 description 1
- 238000005336 cracking Methods 0.000 description 1
- 208000002925 dental caries Diseases 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 231100000518 lethal Toxicity 0.000 description 1
- 230000001665 lethal effect Effects 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 210000000664 rectum Anatomy 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 210000001215 vagina Anatomy 0.000 description 1
Classifications
-
- 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
- G01N33/56944—Streptococcus
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Hematology (AREA)
- Chemical & Material Sciences (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Cell Biology (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The invention discloses a kind of B races streptozyme linked immune assay kit, renovate the box body (2) of (1) including top tape, the sample process area (4) separated by plunger (3), the first cleaning area (5), detection antigen-binding site (6), the second cleaning area (7), luminous substrate land (8), the 3rd cleaning area (9) and terminator (10) are sequentially provided with box body (2)；The plunger (3) is provided with plunger hole；The sample process area (4) is provided with sample treatment solution, sample treatment solution provided with metal stirrer and is coated with the submicron order superparamagnetic immunomagnetic beads of GBS antibody.The present invention can integration complete each steps of GBS immune detections, be not easily introduced pollution, operation is more convenient, and accuracy of detection is higher.
Description
Technical field
The present invention relates to a kind of B races streptozyme linked immune assay kit, belong to technical field of medical detection.
Background technology
B races streptococcus (group B streptococcus, abbreviation GBS), is normally lodged in vagina and rectum, it is one
Conditioned pathogen is planted, normally healthy population infects GBS and not pathogenic.
About 10%~30% pregnant woman has infection GBS according to statistics, wherein 40%~70% can pass in the progress of labor
Neonate.If early stage invasive infection occurs with this bacterium, about 1%~3% in neonate, wherein thering is 5% can lead
It is lethal to die.Therefore, GBS detection is particularly important.Wherein immunity test is a kind of conventional GBS detection methods, but existing
GBS immunologic detection methods in, due to being related to different processing in each step of immune detection, usual sample is needed through excessive
Individual equipment carries out the processing of different step respectively, and detection is more bothered, and when carrying out different step, transfer is also needed to sometimes
Sample is to different carriers (test tube), and this process is readily incorporated pollution, influences accuracy of detection.
The content of the invention
It is an object of the present invention to provide a kind of B races streptozyme linked immune assay kit.It can integration completion
Each step of GBS immune detections, pollution is not easily introduced, and operation is more convenient, and accuracy of detection is higher.
Technical scheme：A kind of B races streptozyme linked immune assay kit, is characterized in：Including top tape
Be sequentially provided with the box body renovated, box body by the sample process area of plunger separates, the first cleaning area, detection antigen-binding site,
Second cleaning area, luminous substrate land, the 3rd cleaning area and terminator；The plunger is provided with plunger hole；At the sample
Manage the submicron order superparamagnetic that area was provided with sample treatment solution, sample treatment solution provided with metal stirrer and be coated with GBS antibody
Immunomagnetic beads；The combination liquid of the GBS antibody containing biotin labeling is provided with the detection antigen-binding site；Luminous substrate land
It is interior to be provided with the substrate nitrite ion containing SA-PE (Streptavidin phycoerythrin)；Terminate liquid is provided with terminator.
In above-mentioned B races streptozyme linked immune assay kit, the component of the sample treatment solution is
| NaCl | 138mM |
| KCl | 2.7mM |
| BSA (bovine serum albumin(BSA)) | 1% mass percent |
In foregoing B races streptozyme linked immune assay kit, the diameter of the submicron order superparamagnetic immunomagnetic beads
For 0.5~10uM, the concentration in sample treatment solution is 50~200mg/ml.
In foregoing B races streptozyme linked immune assay kit, first cleaning area, the second cleaning area and the 3rd are clear
Wash and cleaning fluid is provided with area, the component of cleaning fluid is
| NaCl | 138mM |
| KCl | 2.7mM |
| ddH2O | Solvent |
In the foregoing enzyme-linked immune detection kit of B races streptococcus, the component of the combination liquid is
| NaCl | 138mM |
| KCl | 2.7mM |
| BSA | 1% mass percent |
| The GBS antibody of biotin labeling | 4ug/ml |
| ddH2O | Solvent |
In the foregoing enzyme-linked immune detection kit of B races streptococcus, the component of the substrate nitrite ion is
| NaCl | 138mM |
| KCl | 2.7mM |
| BSA | 1% mass percent |
| SA-PE | 4ug/ml |
| ddH2O | Solvent |
In the foregoing enzyme-linked immune detection kit of B races streptococcus, the component of the terminate liquid is
| NaCl | 138mM |
| KCl | 2.7mM |
| BSA | 1% mass percent |
| ddH2O | Solvent |
In foregoing B races streptozyme linked immune assay kit, the plunger at one end is provided with spring, and the other end is with stretching out
Push rod outside box body；The downside of the push rod outer end is provided with slope.
In foregoing B races streptozyme linked immune assay kit, the taper of 3~5 ° of the plunger hole band, central diameter
For 3~5mm, such setting not only improves magnetic bead and passed through, and blocks each interval liquid freely to wear using capillarity
Cross；Sample process area bottom is closing in wide at the top and narrow at the bottom, and each closing in side presss from both sides 25 °~35 ° angles with vertical direction, in order to
Fully cracking and magnetic bead are smoothly convergeed in plunger hole.
Compared with prior art, the present invention is separated out multiple interval (cavitys) using plunger in same kit, respectively
Individual interval can set the material needed for different detecting steps respectively, and can be with the specific binding of examination target thing using being coated with
The submicron order superparamagnetic immunomagnetic beads of antibody or antigen as examination target thing mobile vehicle, can by using when
The plunger hole of connection, reaction is combined into each interval, therefore whole detection process can be in the case of controllable same
Step in individual kit (with the use of an equipment) needed for all GBS immune detections of progress, it is not easy to produce secondary pollution,
And operating efficiency is greatly improved, accuracy of detection is also greatly improved, and can more solve immune detection need it is many with being arranged
Standby the problem of.The present invention using plunger due to being separated, and it can turn on each area by simple mechanical action
Between, (isolated according to materials such as paraffin without other processing, turn on and heating is needed when assembling, heating influences whether reagent
Reagent and sample in box, carry out influence accuracy of detection), in operation for it is more convenient, and the assembling of plunger it is more simple,
It is easy to the assembling of reagent in each interval.
Brief description of the drawings
Fig. 1 is the structural representation of kit of the present invention.
Mark in accompanying drawing for：1- is renovated, 2- box bodys, 3- plungers, 4- sample process area, the cleaning areas of 5- first, 6- detections
Land, the cleaning areas of 7- second, 8- luminous substrates land, the cleaning areas of 9- the 3rd, 10- terminators, 11- spare areas, 12- bullets
Spring.
Embodiment
The present invention is further illustrated with reference to the accompanying drawings and examples, but be not intended as to the present invention limit according to
According to.
Embodiment.A kind of B races streptozyme linked immune assay kit, as shown in Figure 1：Renovated including top tape 1 box
The sample process area 4 separated by plunger 3, two the first cleaning areas 5, detection antigen-binding sites are sequentially provided with body 2, box body 2
6th, two the second cleaning areas 7, luminous substrate lands 8, two the 3rd cleaning areas 9 and terminators 10；The plunger 3 is provided with post
Consent；The sample process area 4 is provided with sample treatment solution, sample treatment solution provided with metal stirrer and is coated with GBS antibody
Submicron order superparamagnetic immunomagnetic beads；The combination of the GBS antibody containing biotin labeling is provided with the detection antigen-binding site 6
Liquid；The substrate nitrite ion containing SA-PE is provided with luminous substrate land 8；Terminate liquid is provided with terminator 10, terminator 10 may be used also
To connect a spare area.
The component of the sample treatment solution is
| NaCl | 138mM |
| KCl | 2.7mM |
| BSA | 1% mass percent |
| ddH2O | Solvent |
A diameter of 0.5~10uM of the submicron order superparamagnetic immunomagnetic beads, the concentration in sample treatment solution is 50
~200mg/ml.
Cleaning fluid is provided with first cleaning area 5, the second cleaning area 7 and the 3rd cleaning area 9, the component of cleaning fluid is
| NaCl | 138mM |
| KCl | 2.7mM |
| ddH2O | Solvent |
The component of the combination liquid is
| NaCl | 138mM |
| KCl | 2.7mM |
| BSA | 1% mass percent |
| The GBS antibody of biotin labeling | 4ug/ml |
| ddH2O | Solvent |
The component of the substrate nitrite ion is
| NaCl | 138mM |
| KCl | 2.7mM |
| BSA | 1% mass percent |
| SA-PE | 4ug/ml |
| ddH2O | Solvent |
The component of the terminate liquid is
| NaCl | 138mM |
| KCl | 2.7mM |
| BSA | 1% mass percent |
| ddH2O | Solvent |
Described one end of plunger 3 is provided with spring 12, and the other end is with stretching out the push rod outside box body 2；The downside of the push rod outer end
Provided with slope.
The taper of 3~5 ° of the plunger hole band, central diameter is 3~5mm, and the bottom of sample process area 4 is under upper width
Narrow closing in, each closing in side presss from both sides 25 °~35 ° angles with vertical direction.
The operation principle of the kit of the present invention：
1. box body 2 is put samples into, lid 1 is covered, the kit that box body 2 then is inserted into supporting detection device is accommodated
Blend stop is provided with groove, kit holding tank, during insertion, the push rod 8 of each plunger 3 is promoted by blend stop successively so that each
Individual plunger 3 overcomes the elastic force of spring 7 to be moved, and causes plunger hole alignment separation cavity, turns on each interval.
2. metal stirrer and then by electromagnetic coil array is driven, sample is stirred evenly, GBS antigens and sub-micro in sample
GBS antibody on meter level superparamagnetic immunomagnetic beads (hereinafter referred to as carrier) is combined；
3. carrier elutes impurity by the first cleaning area 5；
4. carrier enters on the GBS antigens in detection land 6, the GBS antibody bindings to carrier of biotin labeling；
5. carrier passes through the second cleaning area 7, the GBS antibody of the uncombined biotin labeling of elution；
6. the biotin that carrier enters on luminous substrate land 8, the streptavidin and carrier of luminous substrate (SA-PE)
Specific bond；
7. carrier passes through the 3rd cleaning area 9, the uncombined luminous substrate of elution；
8. carrier enters the luminous substrate phycoerythrin on terminate liquid, carrier and produced in the case where instrument excites light action stably
Be excited light；
9. external equipment is popped one's head in using optical detection and obtains fluorescence intensity signals from the transparent window of terminator, is believed with this
Number the GBS antigens in sample are qualitatively or quantitatively detected.
Claims (9)
1. a kind of B races streptozyme linked immune assay kit, it is characterised in that：The box body (2) of (1) is renovated including top tape,
The sample process area (4) separated by plunger (3), the first cleaning area (5), detection antigen-binding site are sequentially provided with box body (2)
(6), the second cleaning area (7), luminous substrate land (8), the 3rd cleaning area (9) and terminator (10)；Set on the plunger (3)
There is plunger hole；The sample process area (4) is provided with sample treatment solution, sample treatment solution provided with metal stirrer and is coated with
The submicron order superparamagnetic immunomagnetic beads of GBS antibody；The GBS containing biotin labeling is provided with the detection antigen-binding site (6)
The combination liquid of antibody；The substrate nitrite ion containing SA-PE is provided with luminous substrate land (8)；Provided with termination in terminator (10)
Liquid.
2. B races streptozyme linked immune assay kit according to claim 1, it is characterised in that the sample process
The component of liquid is
3. B races streptozyme linked immune assay kit according to claim 1, it is characterised in that：The submicron order
A diameter of 0.5~10uM of superparamagnetic immunomagnetic beads, the concentration in sample treatment solution is 50~200mg/ml.
4. B races streptozyme linked immune assay kit according to claim 1, it is characterised in that first cleaning
Cleaning fluid is provided with area (5), the second cleaning area (7) and the 3rd cleaning area (9), the component of cleaning fluid is
5. B races streptozyme linked immune assay kit according to claim 1, it is characterised in that the combination liquid
Component is
6. B races streptozyme linked immune assay kit according to claim 1, it is characterised in that the substrate colour developing
The component of liquid is
7. B races streptozyme linked immune assay kit according to claim 1, it is characterised in that the terminate liquid
Component is
8. B races streptozyme linked immune assay kit according to claim 1, it is characterised in that：The plunger (3) one
End is provided with spring (12), and the other end is with stretching out the push rod of box body (2) outside；The downside of the push rod outer end is provided with slope.
9. B races streptozyme linked immune assay kit according to claim 1, it is characterised in that：The plunger hole band 3
~5 ° of taper, central diameter is 3~5mm, and sample process area (4) bottom is closing in wide at the top and narrow at the bottom, side of each closing up
25 °~35 ° angles are pressed from both sides with vertical direction.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710427971.8A CN107192823B (en) | 2017-06-08 | 2017-06-08 | Group B streptococcus enzyme-linked immunosorbent assay kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710427971.8A CN107192823B (en) | 2017-06-08 | 2017-06-08 | Group B streptococcus enzyme-linked immunosorbent assay kit |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107192823A true CN107192823A (en) | 2017-09-22 |
| CN107192823B CN107192823B (en) | 2024-03-19 |
Family
ID=59877382
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710427971.8A Active CN107192823B (en) | 2017-06-08 | 2017-06-08 | Group B streptococcus enzyme-linked immunosorbent assay kit |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107192823B (en) |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060003336A1 (en) * | 2004-06-30 | 2006-01-05 | Kimberly-Clark Worldwide, Inc. | One-step enzymatic and amine detection technique |
| CN101533019A (en) * | 2009-04-28 | 2009-09-16 | 中国检验检疫科学研究院 | New method for detecting plague antibodies in serum sample and product thereof |
| CN102112878A (en) * | 2008-06-06 | 2011-06-29 | 国立大学法人富山大学 | Device for detection of influenza virus |
| CN103154739A (en) * | 2010-08-05 | 2013-06-12 | 雅培医护站股份有限公司 | Magnetic immunosensor and method of use |
| CN104316683A (en) * | 2014-10-14 | 2015-01-28 | 南昌大学 | Whole blood-oriented ovarian carcinoma cell detection kit |
| CN104673625A (en) * | 2015-02-13 | 2015-06-03 | 西安交通大学 | Automatic reaction device and method for pretreating cells |
| CN105779398A (en) * | 2015-01-12 | 2016-07-20 | 北京亿森宝生物科技有限公司 | Immunomagnetic bead purification kit and method for 12 kinds of swine common viruses and germs |
| US20160354773A1 (en) * | 2015-06-05 | 2016-12-08 | Yongmei Li | Component of a device, a device, and a method for purifying and testing biomolecules from biological samples |
-
2017
- 2017-06-08 CN CN201710427971.8A patent/CN107192823B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060003336A1 (en) * | 2004-06-30 | 2006-01-05 | Kimberly-Clark Worldwide, Inc. | One-step enzymatic and amine detection technique |
| CN102112878A (en) * | 2008-06-06 | 2011-06-29 | 国立大学法人富山大学 | Device for detection of influenza virus |
| CN101533019A (en) * | 2009-04-28 | 2009-09-16 | 中国检验检疫科学研究院 | New method for detecting plague antibodies in serum sample and product thereof |
| CN103154739A (en) * | 2010-08-05 | 2013-06-12 | 雅培医护站股份有限公司 | Magnetic immunosensor and method of use |
| CN104316683A (en) * | 2014-10-14 | 2015-01-28 | 南昌大学 | Whole blood-oriented ovarian carcinoma cell detection kit |
| CN105779398A (en) * | 2015-01-12 | 2016-07-20 | 北京亿森宝生物科技有限公司 | Immunomagnetic bead purification kit and method for 12 kinds of swine common viruses and germs |
| CN104673625A (en) * | 2015-02-13 | 2015-06-03 | 西安交通大学 | Automatic reaction device and method for pretreating cells |
| US20160354773A1 (en) * | 2015-06-05 | 2016-12-08 | Yongmei Li | Component of a device, a device, and a method for purifying and testing biomolecules from biological samples |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107192823B (en) | 2024-03-19 |
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