CN107727853A - Vibrio splindidus colloidal gold immuno-chromatography test paper strip and preparation method - Google Patents
Vibrio splindidus colloidal gold immuno-chromatography test paper strip and preparation method Download PDFInfo
- Publication number
- CN107727853A CN107727853A CN201710795942.7A CN201710795942A CN107727853A CN 107727853 A CN107727853 A CN 107727853A CN 201710795942 A CN201710795942 A CN 201710795942A CN 107727853 A CN107727853 A CN 107727853A
- Authority
- CN
- China
- Prior art keywords
- flgt
- flagellin
- vibrio splindidus
- vibrio
- monoclonal antibodies
- 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
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
-
- 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
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
-
- 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
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
-
- 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
- G01N2333/28—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Vibrionaceae (F)
Abstract
The present invention disclose one kind can quick detection stichopus japonicus " change skin sick " pathogen --- colloidal gold immuno-chromatography test paper strip and preparation method of Vibrio splindidus, there is bottom plate, on bottom plate sample pad is stained with from one end is from top to bottom stepped successively, gold standard pad, nitrocellulose filter and adsorptive pads, the Vibrio splindidus flagellin FlgT monoclonal antibodies of colloid gold label are coated with the gold standard pad, the nature controlling line b for having the detection line a of coating Vibrio splindidus polyclonal antibody on nitrocellulose filter and being made up of sheep anti-mouse igg, the Vibrio splindidus flagellin FlgT monoclonal antibodies are that mouse is immunized by the hybridoma 4A5 that preserving number is CGMCC No.14311, mouse ascites are purified is prepared.
Description
Technical field
Can quick detection the invention belongs to the immunology rapid detection technical field of aquatic livestock pathogenic bacteria, especially one kind
The colloidal gold immuno-chromatography test paper strip and preparation method of stichopus japonicus " changing skin disease " pathogen-Vibrio splindidus.
Background technology
Vibrio splindidus is under the jurisdiction of category vibrionaceae（Vibrionaceae）, vibrio（Vibrio）, it is stichopus japonicus
（Apostichopus japonicus）" changing skin disease "（Skin ulcer syndrome）Main Pathogenic Bacteria.The disease has that infectiousness is strong, passes
The characteristics of scope is wide is broadcast, the symptom such as the body surface ulcer of stichopus japonicus occurrence of large-area, mouth turgescence canker, body atrophy, row can be caused dirty.This
Outside, Vibrio splindidus can result in a variety of fish, bivalve shellfish, shell-fish and echinoderm morbidity even death, make infected
Fish body both sides, especially proximate to there is blutpunkte at afterbody, cause skin to rot, form ulcer, dorsal fin, pectoral fin, the fin of tail fin
Bar base portion is congested, the symptom such as kidney, liver erosion.At present, mainly there is traditional microorganism for the detection method of Vibrio splindidus
Learn detection technique, immunofluorescence technique, enzyme linked immunosorbent assay and PCR technologies etc..Above-mentioned detection method because complex operation,
Time-consuming instrument and equipment long, needs are special and professional, and do not reach the quick, purpose of Site Detection, be not suitable in basic unit
Promote the use of.Colloidal gold immuno-chromatography test paper strip with its is fast and convenient, be not required to specific apparatus, result can naked eyes interpretation, sensitivity
The advantages that height, high specificity, it has also become one of current quick sensitive immunoassay technology, and start gradually to support applied to aquatic products
Grow the multiple fields of industry.
Colloidal gold immuno-chromatography test paper strip is the test strips that width is 0.3 cm, has bottom plate, on bottom plate from one end successively
It is stained with sample pad, gold standard pad, nitrocellulose filter and adsorptive pads from top to bottom steppedly, it is overlapping between each several part to connect.
The antibody of colloid gold label is generally coated with gold standard pad, has the detection line a of coated antibody on nitrocellulose filter and has sheep
The nature controlling line b that anti-mouse IgG is formed.Its Cleaning Principle is the quilt being placed in sample pad using the capillarity of adsorptive pads formation
Detect antibody binding of the antigen first with the colloid gold label in gold standard pad, and along nitrocellulose filter（NC films）Continue to
Preceding movement, when arrival is fixed with the detection line a of antibody, antibody is captured, and as reaction is carried out, constantly enrichment reaches meat
Eye visible horizon；The antibody of excessive colloid gold label continues to move along, and reaches the nature controlling line b for being fixed with sheep anti-mouse igg
When, it is captured and is constantly enriched with.If therefore sample is the positive, occurs vitta band in detection line and nature controlling line, if sample
For feminine gender, then only occurs the vitta band of same color on nature controlling line.But due to not being applied to collaurum so far
The Vibrio splindidus monoclonal antibody of immuno-chromatographic test paper strip, so that still using microbiologic inhibition tests for Vibrio splindidus at present
Technology, immunofluorescence technique, enzyme linked immunosorbent assay, bacterial agglutination method and PCR technologies etc. are detected, colloid gold immune
Chromatograph test strip is not applied in Vibrio splindidus is detected.
The content of the invention
The present invention is to solve the above-mentioned technical problem present in prior art, there is provided one kind can quick detection stichopus japonicus
The colloidal gold immuno-chromatography test paper strip and preparation method of " changing skin disease " pathogen-Vibrio splindidus.
The present invention technical solution be：A kind of Vibrio splindidus colloidal gold immuno-chromatography test paper strip, has bottom plate, in bottom plate
On from one end is from top to bottom stepped be successively stained with sample pad, gold standard pad, nitrocellulose filter and adsorptive pads, the gold mark
The Vibrio splindidus flagellin FlgT monoclonal antibodies of colloid gold label are coated with pad, have coating bright on nitrocellulose filter
The detection line a of the rotten vibrios polyclonal antibody and nature controlling line b being made up of sheep anti-mouse igg, the Vibrio splindidus flagellin
FlgT monoclonal antibodies are resisted by the anti-Vibrio splindidus flagellin FlgT monoclonals that preserving number is CGMCC No. 14311
Mouse is immunized in the hybridoma cell strain 4A5 of body, obtains that mouse ascites are purified to be prepared.
A kind of preparation method of above-mentioned Vibrio splindidus colloidal gold immuno-chromatography test paper strip, it is characterised in that according to the following steps
Carry out：
A. Vibrio splindidus flagellin FlgT monoclonal antibodies are prepared：Take the anti-brilliance that preserving number is CGMCC No. 14311
Mouse is immunized in the hybridoma cell strain 4A5 of vibrios flagellin FlgT monoclonal antibodies, takes mouse ascites, puts centrifuge tube centrifugation
Supernatant is collected afterwards and is purified, and produces Vibrio splindidus flagellin FlgT monoclonal antibodies；
B. collaurum is prepared：30 nm colloidal gold solutions are prepared as reducing agent using trisodium citrate, and it is molten to adjust collaurum
The pH of liquid to 8.2, it is standby；
C. the Vibrio splindidus flagellin FlgT monoclonal antibodies of colloid gold label are prepared：By magnificent arc under magnetic stirrer
Bacterium flagellin FlgT monoclonal antibodies are slowly added in standby colloidal gold solution, the quality and collaurum of monoclonal antibody
The volume ratio of solution is 0.018：1, the Vibrio splindidus flagellin FlgT that colloid gold label is formed after purified and concentration is mono-
Clonal antibody；
D. to gold standard pad and nitrocellulose film process：The Vibrio splindidus flagellin FlgT monoclonals of colloid gold label are resisted
Body is with 9 μ L/cm2Even application is in gold standard pad；Vibrio splindidus polyclonal antibody is diluted to 0.5 mg/mL, with 1 μ L/cm
It is sprayed on nitrocellulose filter, forms detection line a；Sheep anti-mouse igg is diluted to 1 mg/mL, is sprayed at 1 μ L/cm
On nitrocellulose filter, nature controlling line b, 37 DEG C of 4 h of drying are formed；
E. assemble：It is fixed on sample pad, gold standard pad, nitrocellulose filter and adsorptive pads are stepped from top to bottom successively not
On dry glue bottom plate, there is lap between sample pad, gold standard pad, nitrocellulose filter and adsorptive pads each several part, it is wide to be cut into specification
The test strips of degree.
The a steps are that the anti-Vibrio splindidus flagellin FlgT monoclonals for taking preserving number CGMCC No. 14311 resist
Cell suspension is made in the hybridoma cell strain 4A5 of body, and 1000 r/min centrifuge 5 min, supernatant removed, with 1640 cell culture fluids
Cell is resuspended, it is 5 × 10 to make final cell density6Cells/mL, obtain hybridoma liquid；It is immunized with hybridoma liquid
Mouse, mouse ascites are taken after 7 ~ 10 days, put centrifuge tube centrifugation, 2000 r/min, l0 min, collected after centrifugation supernatant simultaneously use
The ascites of Protein-G affinity chromatography purified pools, produce Vibrio splindidus flagellin FlgT monoclonal antibodies.
Present invention can apply to detect Vibrio splindidus, compared with existing detection method, there is advantages below：
（1）Detection is quick：Detection time only 5 ~ 10 min, scene can go out result；
（2）High specificity, high sensitivity：This test strips and the common pathogenic bacteria no cross reaction of other aquatic livestocks, minimum inspection
Survey is limited to 1 × 105 cells/mL ；
（3）It is easy to operate, without professional and the specific apparatus of auxiliary；
（4）The term of validity is grown：2-8 DEG C is sealed, and the term of validity is 1 year.
Brief description of the drawings
Fig. 1 is the structural representation of the embodiment of the present invention.
Hybridoma 4A5 preservation dates：On July 5th, 2017；
Classification And Nomenclature：The hybridoma cell strain of anti-Vibrio splindidus flagellin FlgT monoclonal antibodies；
Depositary institution：China Committee for Culture Collection of Microorganisms's common micro-organisms center（CGMCC）；
Depositary institution address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3；
Preserving number：CGMCC No. 14311 .
Embodiment
The Vibrio splindidus colloidal gold immuno-chromatography test paper strip structure of the present invention is same as the prior art, has bottom plate 5, the bottom of at
On plate 5 sample pad 4, gold standard pad 3, nitrocellulose filter 2 and adsorptive pads 1 are stained with from one end is stepped from top to bottom successively
, it is overlapping between each several part to connect.With prior art except that being coated with the Vibrio splindidus of colloid gold label in gold standard pad 3
Flagellin FlgT monoclonal antibodies, have on nitrocellulose filter 2 coating Vibrio splindidus polyclonal antibody detection line a and
There is the nature controlling line b that sheep anti-mouse igg is formed, the Vibrio splindidus flagellin FlgT monoclonal antibodies are to be by preserving number
Mouse is immunized in CGMCC No. 14311 hybridoma 4A5, obtains that mouse ascites are purified to be prepared.
The Vibrio splindidus colloidal gold immuno-chromatography test paper strip preparation method of the present invention, is carried out according to the following steps：
1. prepare Vibrio splindidus flagellin FlgT monoclonal antibodies：Take and grow vigorous, the good hybridoma 4A5 of form
Cell suspension is made, 1000 r/min centrifuge 5 min, remove supernatant, and cell is resuspended with 1640 cell culture fluids, makes final cell
Density is 5 × 106Cells/mL, it is standby；Take 6 ~ 8 week old Balb/C small white mouses, the sterile atoleine of intraperitoneal injection
Only, after 1 week, the mL/ of intraperitoneal injection hybridoma cell strain 4A5 0.5 only, after 7 ~ 10 days, are shown in that mouse web portion is obvious to 0.5 mL/
Expand, draw neck to put to death mouse, after sterilizing lower abdomen skin with cotton ball soaked in alcohol, extract ascites with syringe, the ascites of collection is mixed
Close, put centrifuge tube centrifugation（2000 r/min, l0 min）, collected after centrifugation supernatant is pure using Protein-G affinity chromatographies
Change the ascites collected, i.e. Vibrio splindidus flagellin FlgT monoclonal antibodies.
2. prepare collaurum：30 nm collaurums are prepared using trisodium citrate reduction method, are by the chlorine of 100 mL 0.01%
Change gold, be heated to seething with excitement；2 mL 1% sodium citrate is taken to add in above-mentioned solution, it is rapid well mixed, then keep boiling 10
Min, until solution colour is in bright red, distilled water recovers to l00 mL volumes after natural cooling；Colloidal gold solution brown
4 DEG C of preservations of bottle, standby, the uniformity and particle diameter of observing colloid gold grain under transmission electron microscope.
3. prepare the Vibrio splindidus flagellin FlgT monoclonal antibodies of colloid gold label：Take the collaurum prepared
The mL of solution 10, with 0.2 mol/L K2CO3Solution adjusts pH value of solution to 8.2, under electromagnetic agitation, adds 180 while stirring
The mg/mL monoclonal antibodies of μ L 1, continue to stir 30 min, then 5% BSA is added dropwise to final concentration of 1%, be added dropwise 5%
PEG 20000 to final concentration of 1%（With the residual epitope of stable colloid gold grain）, stir 30 min.1500 r/min, 4 DEG C
20 min are centrifuged, take supernatant；By supernatant with 10000 r/min, 4 DEG C of 60 min of centrifugation, supernatant is abandoned；It will precipitate with 10 mL gold
Mark redissolves liquid and is resuspended, repeated centrifugation 2~3 times；Finally precipitation is dissolved in the gold medal mark dilution of original volume 1/10,4 DEG C of preservations are standby
With.
4. pair gold standard pad and nitrocellulose film process：By the Vibrio splindidus flagellin FlgT Dan Ke of colloid gold label
Grand antibody is with 9 μ L/cm2Even application is in gold standard pad；Vibrio splindidus polyclonal antibody is diluted to 0.5 mg/mL, with 1 μ
L/cm is sprayed on nitrocellulose filter, forms detection line a；Sheep anti-mouse igg is diluted to 1 mg/mL, sprayed with 1 μ L/cm
It is applied on nitrocellulose filter, forms nature controlling line b, 37 DEG C of 4 h of drying.
5. assembling：By sample pad, gold standard pad, nitrocellulose filter and adsorptive pads successively stepped fixation from top to bottom
In on adhesive sticker bottom plate, there is lap between sample pad, gold standard pad, nitrocellulose filter and adsorptive pads each several part, be cut into rule
The test strips of lattice width.
The Vibrio splindidus colloidal gold immuno-chromatography test paper strip result judgement of the present invention：
Test strips are inserted in sample suspension and taken out after 10 s, room temperature places 5 ~ 10 min, observes nature controlling line and detection line colour developing
Situation, respectively occurs 1 red stripes at positive reaction detection line and nature controlling line；There is a red in negative reaction nature controlling line
Band；If nature controlling line occurs without red stripes, then it represents that test-strips fail.
The specificity analysis of the Vibrio splindidus colloidal gold immuno-chromatography test paper strip of the present invention：
1. the specificity analysis of test strips
With the present invention Vibrio splindidus colloidal gold immuno-chromatography test paper strip detection Vibrio splindidus (Vibrio splendidus)
ATCC33125, Vibrio splindidus (Vibrio splendidus) KCTC12679, Vibrio splindidus (Vibrio splendidus)
2CLM002, vibrio parahaemolytious (Vibrio parahaemolyticus) KCTC2471, Vibrio harveyi (Vibrio harveyi) ATCC14126, vibrio alginolyticus (Vibrio alginolyticus) KCCM40513, Vibrio anguillarum (Vibrio anguillarum) KCTC2711, vibrio fluvialis (Vibrio fluvialis) KCCM40827, Vibrio vulnificus (Vibrio vulnificus) ATCC2126, Edwardsiella tarda (Edwardsiella tarda) ATCC15947, Shewanella
(Shewanella oneidensis) KCCM41821, while using PBS as negative control, evaluation specificity and cross reaction feelings
Condition, bacterial concentration used are 108 cells/mL.As a result show, with the ELISA test strip vibrio parahaemolytious after assembling, breathe out arc maintenance
Bacterium, vibrio alginolyticus, Vibrio anguillarum, vibrio fluvialis, Vibrio vulnificus, Edwardsiella tarda, the result of Shewanella are feminine gender, and
The testing result of 3 plants of Vibrio splindidus illustrates this test strips and test strain no cross reaction, high specificity into the positive.
2. the sensitivity technique of test strips
Take various concentrations（1×109 Cells/mL to 1 × 104 cells/mL）Vibrio splindidus bacteria suspension, tried with collaurum
Paper slip detects, and evaluates its detection limit.As a result show that the lowest detection of test strips is limited to 1 × 105 cells/mL。
3. the stability test of test strips
Test strips are sealed 1 year in 2-8 DEG C, every detection in 1 month once, indices meet requirements above.
Claims (3)
1. a kind of magnificent bacterium colloidal gold immuno-chromatography test paper strip, there is bottom plate（5）, in bottom plate（5）On from one end successively from top to bottom
Sample pad is stained with steppedly（4）, gold standard pad（3）, nitrocellulose filter（2）And adsorptive pads（1）, it is characterised in that：It is described
Gold standard pad（3）On be coated with the Vibrio splindidus flagellin FlgT monoclonal antibodies of colloid gold label, nitrocellulose filter（2）
On have the detection line a for the being coated with Vibrio splindidus polyclonal antibody and nature controlling line b being made up of sheep anti-mouse igg, the magnificent arc
Bacterium flagellin FlgT monoclonal antibodies are that mouse is immunized by the hybridoma 4A5 that preserving number is CGMCC No. 14311,
Mouse ascites are purified is prepared.
2. a kind of preparation method of Vibrio splindidus colloidal gold immuno-chromatography test paper strip as described in claim 1, its feature exist
Carried out according to the following steps：
A. Vibrio splindidus flagellin FlgT monoclonal antibodies are prepared：Take the anti-brilliance that preserving number is CGMCC No. 14311
Mouse is immunized in the hybridoma cell strain 4A5 of vibrios flagellin FlgT monoclonal antibodies, takes mouse ascites, puts centrifuge tube centrifugation
Supernatant is collected afterwards and is purified, and produces Vibrio splindidus flagellin FlgT monoclonal antibodies；
B. collaurum is prepared：Using 30 nm colloidal gold solutions, and the pH to 8.2 of colloidal gold solution is adjusted, it is standby；
C. the Vibrio splindidus flagellin FlgT monoclonal antibodies of colloid gold label are prepared：By magnificent arc under magnetic stirrer
Bacterium flagellin FlgT monoclonal antibodies are slowly added in standby colloidal gold solution, the quality and collaurum of monoclonal antibody
The volume ratio of solution is 0.018：1, the Vibrio splindidus flagellin FlgT Dan Ke of colloid gold label are formed after purified and concentration
Grand antibody；
D. to gold standard pad and nitrocellulose film process：The Vibrio splindidus flagellin FlgT monoclonals of colloid gold label are resisted
Body is with 9 μ L/cm2Even application is in gold standard pad；Vibrio splindidus polyclonal antibody is diluted to 0.5 mg/mL, with 1 μ L/cm
It is sprayed on nitrocellulose filter, forms detection line a；Sheep anti-mouse igg is diluted to 1 mg/mL, is sprayed at 1 μ L/cm
On nitrocellulose filter, nature controlling line b, 37 DEG C of 4 h of drying are formed；
E. assemble：It is fixed on sample pad, gold standard pad, nitrocellulose filter and adsorptive pads are stepped from top to bottom successively not
On dry glue bottom plate, there is lap between sample pad, gold standard pad, nitrocellulose filter and adsorptive pads each several part, it is wide to be cut into specification
The test strips of degree.
3. the preparation method of Vibrio splindidus colloidal gold immuno-chromatography test paper strip according to claim 2, it is characterised in that institute
State the hybridization that a steps are the anti-Vibrio splindidus flagellin FlgT monoclonal antibodies for taking preserving number to be CGMCC No. 14311
Cell suspension is made in tumor cell strain 4A5, and 1000 r/min centrifuge 5 min, remove supernatant, is resuspended with 1640 cell culture fluids thin
Born of the same parents, it is 5 × 10 to make final cell density6Cells/mL, obtain hybridoma liquid；With hybridoma liquid be immunized mouse, 7 ~
Mouse ascites are taken after 10 days, put centrifuge tube centrifugation, 2000 r/min, l0 min, collected after centrifugation supernatant simultaneously use Protein-
The ascites of G affinity chromatography purified pools, produce Vibrio splindidus flagellin FlgT monoclonal antibodies.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710795942.7A CN107727853B (en) | 2017-09-06 | 2017-09-06 | Vibrio splindidus colloidal gold immuno-chromatography test paper strip and preparation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710795942.7A CN107727853B (en) | 2017-09-06 | 2017-09-06 | Vibrio splindidus colloidal gold immuno-chromatography test paper strip and preparation method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107727853A true CN107727853A (en) | 2018-02-23 |
| CN107727853B CN107727853B (en) | 2019-05-10 |
Family
ID=61204938
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710795942.7A Active CN107727853B (en) | 2017-09-06 | 2017-09-06 | Vibrio splindidus colloidal gold immuno-chromatography test paper strip and preparation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107727853B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109030823A (en) * | 2018-10-15 | 2018-12-18 | 中国烟草总公司郑州烟草研究院 | A kind of colloidal gold strip and its preparation method and application detecting tobacco ralstonia solanacearum |
| CN109187970A (en) * | 2018-12-05 | 2019-01-11 | 鲁东大学 | It is a kind of quickly to detect aquatic products disease gold mark nucleic acid test strip and preparation method thereof |
| CN111171138A (en) * | 2020-01-14 | 2020-05-19 | 大连深蓝肽科技研发有限公司 | Peptide fragment, monoclonal antibody, colloidal gold test strip and detection method for detecting stichopus japonicus oligopeptide |
| CN112710832A (en) * | 2020-12-22 | 2021-04-27 | 扬州大学 | Test strip for detecting avian adenovirus serotype 4 based on Fiber2 protein |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS63216490A (en) * | 1987-03-06 | 1988-09-08 | Sagami Chem Res Center | Production of eicosapentaenoic acid by microorganism and microorganism using therefor |
| EP1170382A2 (en) * | 2000-07-06 | 2002-01-09 | Institut Francais de Recherche pour l'Exploitation de la Mere(Ifremer) | Method for diagnosing a bacterial infection in the crassostrea gigas oyster larvae |
| CN1944681A (en) * | 2006-10-25 | 2007-04-11 | 大连水产学院 | Method for quick detecting vibrio splendidus |
| CN103468806A (en) * | 2013-09-10 | 2013-12-25 | 中国科学院海洋研究所 | Quick detection method for scallop pathogenic vibrio splendidus |
| CN105039547A (en) * | 2015-07-27 | 2015-11-11 | 宁波大学 | PCR detection primer and method of Vibro Splendidus and application of PCR detection primer |
| CN105198988A (en) * | 2015-09-30 | 2015-12-30 | 大连理工大学 | Vibrio-splendidus-resistant egg yolk antibody and preparation method thereof |
| CN106811542A (en) * | 2017-03-28 | 2017-06-09 | 大连海洋大学 | Genetic chip and application method for detecting ginseng shrimp shellfish culture zone morbid vibrio group |
| CN107064490A (en) * | 2017-05-18 | 2017-08-18 | 中国科学院烟台海岸带研究所 | A kind of utilize recombinates detection reagent bar of fluorescence phycobniliprotein subunit and preparation method thereof |
-
2017
- 2017-09-06 CN CN201710795942.7A patent/CN107727853B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS63216490A (en) * | 1987-03-06 | 1988-09-08 | Sagami Chem Res Center | Production of eicosapentaenoic acid by microorganism and microorganism using therefor |
| EP1170382A2 (en) * | 2000-07-06 | 2002-01-09 | Institut Francais de Recherche pour l'Exploitation de la Mere(Ifremer) | Method for diagnosing a bacterial infection in the crassostrea gigas oyster larvae |
| CN1944681A (en) * | 2006-10-25 | 2007-04-11 | 大连水产学院 | Method for quick detecting vibrio splendidus |
| CN103468806A (en) * | 2013-09-10 | 2013-12-25 | 中国科学院海洋研究所 | Quick detection method for scallop pathogenic vibrio splendidus |
| CN105039547A (en) * | 2015-07-27 | 2015-11-11 | 宁波大学 | PCR detection primer and method of Vibro Splendidus and application of PCR detection primer |
| CN105198988A (en) * | 2015-09-30 | 2015-12-30 | 大连理工大学 | Vibrio-splendidus-resistant egg yolk antibody and preparation method thereof |
| CN106811542A (en) * | 2017-03-28 | 2017-06-09 | 大连海洋大学 | Genetic chip and application method for detecting ginseng shrimp shellfish culture zone morbid vibrio group |
| CN107064490A (en) * | 2017-05-18 | 2017-08-18 | 中国科学院烟台海岸带研究所 | A kind of utilize recombinates detection reagent bar of fluorescence phycobniliprotein subunit and preparation method thereof |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109030823A (en) * | 2018-10-15 | 2018-12-18 | 中国烟草总公司郑州烟草研究院 | A kind of colloidal gold strip and its preparation method and application detecting tobacco ralstonia solanacearum |
| CN109030823B (en) * | 2018-10-15 | 2020-10-27 | 中国烟草总公司郑州烟草研究院 | Colloidal gold test strip for detecting ralstonia solanacearum and preparation method and application thereof |
| CN109187970A (en) * | 2018-12-05 | 2019-01-11 | 鲁东大学 | It is a kind of quickly to detect aquatic products disease gold mark nucleic acid test strip and preparation method thereof |
| CN111171138A (en) * | 2020-01-14 | 2020-05-19 | 大连深蓝肽科技研发有限公司 | Peptide fragment, monoclonal antibody, colloidal gold test strip and detection method for detecting stichopus japonicus oligopeptide |
| CN112710832A (en) * | 2020-12-22 | 2021-04-27 | 扬州大学 | Test strip for detecting avian adenovirus serotype 4 based on Fiber2 protein |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107727853B (en) | 2019-05-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107727853A (en) | Vibrio splindidus colloidal gold immuno-chromatography test paper strip and preparation method | |
| Tam et al. | Serological classification of Neisseria gonorrhoeae with monoclonal antibodies | |
| CN101315382A (en) | Immune colloidal gold test paper strip for detecting yapamicin relict and preparation thereof | |
| CN109655435B (en) | Fluorescence quenching test paper for detecting okadaic acid and preparation method and application thereof | |
| JPS60259963A (en) | Multiple section immunometric assay method | |
| CN108956989B (en) | Detection reagent card for helicobacter pylori typing detection and preparation method thereof | |
| JPH02500219A (en) | Detection method | |
| CN102590527B (en) | Method for detecting insecticidal crystal proteins Bt Cry1Ab/Ac and special enzyme linked immunosorbent assay kit thereof | |
| CN106226518A (en) | Canine distemper virus colloidal gold immunochromatographydetection detection test paper bar and preparation method thereof | |
| Holt et al. | Rapid detection of Salmonella enteritidis in pooled liquid egg samples using a magnetic bead-ELISA system | |
| CN113637081A (en) | Hybridoma cell strain secreting pendimethalin-resistant monoclonal antibody and application thereof | |
| CN106556701A (en) | Brucella melitensis indirect ELISA antibody assay kit | |
| CN106282126A (en) | The monoclonal antibody hybridoma cell strain YH2 of one strain preventing from heavy metal cadmium and application thereof | |
| CN106405093A (en) | Detection kit for detecting infectious bovine rhinotracheitis virus antibody | |
| JPS63159762A (en) | Method of evaluating mouth microbe | |
| WO2018095254A1 (en) | Monoclonal cell line c4 capable of secreting monoclonal antibody recognizing methylene blue and application thereof | |
| CN105486871B (en) | A kind of quick detection canine parvovirus antibody blood clotting suppresses colloidal gold strip, kit and the detection method of potency | |
| CN102539768A (en) | EB (Epstein-Barr) virus Zta IgA (Immunoglobulin A) antibody colloidal gold detection kit and preparation method thereof | |
| Coutelier et al. | Rheumatoid factor production in 129/Sv mice: involvement of an intestinal infectious agent. | |
| CN108008127B (en) | Preparation method and application of enterobacter sakazakii colloidal gold test strip | |
| CN102676459A (en) | Monoclonal antibody resistant to thermostable direct hemolysin of vibrio parahaemolyticus and preparation method of monoclonal antibody | |
| CA2078162A1 (en) | Specific anti-salmonella monoclonal reagents, and unique serological approach for the detection of different common serotypes of salmonella and the like | |
| JP2014066641A (en) | Method for detecting escherichia coli o157 | |
| Prevot et al. | A lectin immunotest using erythrocytes as marker (ERYTHRO-LIT) for detection and titration of rotavirus antigen | |
| CN107337730A (en) | A kind of monoclonal antibody of anti-channel catfish virus nucleocapsid albumen and its application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |