CN108226507A - A kind of yellow fever antigen near-infrared fluorescent detection kit and application thereof - Google Patents
A kind of yellow fever antigen near-infrared fluorescent detection kit and application thereof Download PDFInfo
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Abstract
The present invention relates to biotechnology, more particularly to a kind of yellow fever antigen near-infrared fluorescent detection kit and application thereof.The present invention provides a kind of yellow fever antigen near-infrared fluorescent detection kit, including the reagent strip being located on backboard, since reagent strip be followed successively by sample pad being loaded end, be marked with the glass fibre element film of immune fluorescent probe, be coated with the nitrocellulose filter and blotting paper of LP antibody.Detection kit provided by the present invention, can be quick with reference to Near-infrared fluorescent detection technique using the urine of patient or sputum as sample is detected, and accurately detects specificity, solubility LP antigens in patient's sample.
Description
Technical field
The present invention relates to biotechnology, more particularly to a kind of yellow fever antigen near-infrared fluorescent detection kit and
Its purposes.
Background technology
Yellow fever (yellow fever) is a kind of using fever, yellow subcutaneous ulcer, bleeding as the acute viral infection mainly showed
Sick river.Since environmental disruption, international tourism increase and the vaccine coverage of yellow fever epidemic situation country reduces since modern age, 1979
Nian Qi, yellow fever restart some areas outbreak of epidemic in Africa and South America, it is estimated that the whole world actually has every year
200000 patients, death toll is up to 30 000, and only sub-fraction is made a definite diagnosis.Although Kou Qian China has no the big of yellow fever
Scale is popular, but the certain areas geographical conditions in south China are similar to yellow fever region occurred frequently, and exists to yellow fever
The media such as malicious sensitive host animal and Aedes aegypti, there is also the possibility of morbidity, simultaneously because not occurring before China
It is popular on a large scale to cross yellow fever, China resident is relatively low to its immunity, once breaking out will generate a panic.Therefore, it is fast to inquire into it
Fast diagnostic method, epidemic situation monitoring and epidemiological survey to yellow fever virus are important so as to which corresponding control measure be taken to have
Meaning.
Invention content
In view of the foregoing deficiencies of prior art, the purpose of the present invention is to provide a kind of yellow fever antigen near-infrared is glimmering
Light detection kit and application thereof for establishing efficiently feasible yellow fever antigen Near-infrared fluorescent detection technique, solves existing
The problems in technology.
In order to achieve the above objects and other related objects, it is glimmering to provide a kind of yellow fever antigen near-infrared for first aspect present invention
Light detection kit, including the reagent strip being located on backboard, reagent strip be followed successively by since being loaded end sample pad, be marked with it is immune
The glass fibre element film of fluorescence probe, the nitrocellulose filter and blotting paper for being coated with LP antibody.
Preferably, the immune fluorescent probe is the coated near-infrared fluorescent latex beads particle of LP antibody.
It is furthermore preferred that a diameter of 140-160nm of the fluorescent latex microsphere particle.
Near infrared light (NIR), is the electromagnetic wave between visible ray (VIS) and mid-infrared light (MIR), ASTM definition
NIR is electromagnetic wave of the wavelength in the range of 780nm-2526nm.In near-infrared region, organism self-absorption or fluorescence intensity are very
It is small, background interference can be avoided.Simultaneously because the biquadratic of scattered light intensity and wavelength is in inverse ratio, the scattering interference of near infrared region is big
To reduce.
Second aspect of the present invention provides the preparation method of the yellow fever antigen near-infrared fluorescent detection kit, including such as
Lower step:
1) disperse with phosphate buffer by fluorescent latex particles eccentric cleaning and by latex particle (fluorescent latex is micro-
The phosphate buffer solution cleaning of grain PH7.3,0.01mol/L, 10000rpm centrifugation l0min, the sediment after centrifugation
Redissolved with phosphate buffer, and disperseed emulsion particle with ultrasonic cell disruptor, this step is repeated twice), it is cleaning
LP antibody is added in good latex particle;Adding in EDCA makes antibody be coupled with particle;It adds on ethanol amine closing latex particle
Extra group, prepares probe solution;
2) it will be added on nitrocellulose filter containing the spray of the buffer solution of LP antibody using trace of albumin point membranous system, drying is standby
With;
3) with the probe solution sized glass fibres film for preparing, drying for standby in vacuum drier;
4) by the coating nitrocellulose filter of LP antibody, the glass fibre element film for being marked with immune fluorescent probe, sample pad, water suction
Paper and backboard assembling are prepared into yellow fever antigen near-infrared fluorescent detection kit.
Preferably, in the step 1, the phosphoric acid of phosphate buffer 0.009-0.0llmol/L, PH7.25-7.35
Salt buffer.
Preferably, in the step 1, a diameter of 140-160nm of fluorescent latex particles.
Preferably, in the step 1, latex particle, LP antibody, EDCA weight ratio be 9-11:0.9-1. 1；0.9 1
1.1。
In an embodiment of the present invention latex particle, LP antibody, EDCA inventory be respectively 1mg, l00ug, 100ug.
The addition of ethanol amine is appropriate in the step 1, and those skilled in the art can adjust ethyl alcohol according to actual conditions
The dosage of amine.
Preferably, in the step 2, the buffer solution is PBS buffer solution.
Those skilled in the art can choose debita spissitudo and the PBS buffer solution of pH.
Preferably, in the step 2, a concentration of 1. 8-2.2mg/ml of LP antibody in the buffer solution containing LP antibody.
Preferably, in the step 2, the spray dosage of LP antibody is 0.9-1.1ul/cm.
Preferably, in the step 2, the actual conditions of drying are dried for 36-38 DEG C of baking oven.
Preferably, the preparation method of the LP antibody is as follows:
1) preparation of antigen:By yellow fever inoculation on culture medium, physiological saline eluting bacterial from tablet is used after culture,
Centrifugation obtains bacterium mud (while removing the liquid component that tablet is eluted), and bacterium mud adds appropriate PBS buffer solution sonic disruption, it is excessively cloudy from
Sub- column purification is washed with NaCl aqueous solutions, then is eluted with NaCl aqueous solutions, eluent dress bag filter, dense with PEG embedding water suctions
Contracting obtains antigen;
Preferably, the yellow fever bacterial strain is purchased from ATCC.
Preferably, the condition of culture in the preparation process of the antigen is specially:It 37 DEG C, cultivates under conditions of 5% C02,
Lower lawn is washed after 3-5 days.
Preferably, the culture medium in the preparation process of the antigen is buffers active carbon yeast agar medium (BCYE).
Preferably, the condition that the centrifugation obtains bacterium mud is 4500-5500rpm.
Preferably, described when being washed with NaCL aqueous solutions, the solution concentration of NaCl is 45-55mM.
Preferably, during the elution with NaCl aqueous solutions, the solution concentration of NaCl is 135-165mM.
2) preparation of monoclonal antibody:
Animal immune:Mouse is immunized;
Preferably, it is described that immune specifically comprise the following steps is carried out to mouse:Antigen adds the Fu Shi of equivalent complete when immune for the first time
Full adjuvant is fully emulsified, and for the second time and third time antigen adds equivalent freund 's incomplete adjuvant fully emulsified, the preceding abdominal cavity of cell fusion
Direct injection antigen booster immunization, one exempts from antigen Freund Freund's complete adjuvant, and subcutaneous inoculation, amount of antigen is 0.lmg/ mouse, 2 after 15 days
Exempt from, subcutaneous inoculation antigen Freund Freund's incomplete adjuvant, amount of antigen is 0.2mg/ mouse, and three exempt from antigen Freund and not exclusively help after 25 days
Agent, subcutaneous inoculation, amount of antigen are 0.2mg/ mouse, tail are cut after 10 days, blood is taken to survey ELISA potency, choose potency and are more than 100,000 times
Mouse carries out booster immunization, and aqua intraperitoneal injection antigen 1 mg/ mouse take spleen cell to merge after three days;
Cell fusion:Mouse is put to death, sterile working takes out spleen, prepares bone-marrow-derived lymphocyte suspension, by bone-marrow-derived lymphocyte with preparing
The good mixing of the homology SP2/0 myeloma cell in exponential phase, prepares hybridoma, and carry out hybridoma sieve
Choosing;
The specific method of filtering hybridoma is in the preparation process of the preferred monoclonal antibody:It is trained with HAT culture mediums
Cell is supported, the hybridoma of myeloma cell and bone-marrow-derived lymphocyte can be filtered out after 2 weeks.
The cloning of hybridoma:Until 100% antibody positive is detected in each hole of clone cell growth;
Preferably, the method for the cloning is limiting dilution assay.
The preparation of antibody:Paraffin is injected intraperitoneally in mouse first, after 1-2 weeks, intraperitoneal inoculation hybridoma, 1-2 weeks
Afterwards, ascites is extracted with syringe, a large amount of monoclonal antibody can be obtained after purification.Preferably, the intraperitoneal inoculation hybridization
During oncocyte, hybridoma concentration is adjusted to 1 × 105A/ml injects lml inoculations by every mouse peritoneal.
Preferably, in the preparation process of the monoclonal antibody, mouse is 6-8 week old BALB/C mices.
Preferably, the purifying of monoclonal antibody:Selection HiTraprProtein A FF 5m1 prepackage chromatographies resist
Body, the antibody for collecting purifying are spare.
Third aspect present invention provides the yellow fever antigen near-infrared fluorescent detection kit and is detected in yellow fever antigen
The purposes in field.
The purposes is specially that yellow fever antigen is carried out using the yellow fever antigen near-infrared fluorescent detection kit
Detection.
Near infrared light (NIR) is the electromagnetic wave between visible ray (VIS) and mid-infrared light (MIR), ASTM definition NIR
The electromagnetic wave for being wavelength in the range of 780nm-2526nm.In near-infrared region, organism self-absorption or fluorescence intensity very little can
Avoid background interference.Simultaneously because the biquadratic of scattered light intensity and wavelength is in inverse ratio, the scattering interference of near infrared region greatly subtracts
It is few.With the foundation of laser fluorescence, sensor, immunoassay device, near-infrared fluorescent analyzer is shown in immunoassays field
Great superiority is shown.
Detection kit provided by the present invention is using the urine of patient or sputum as detection sample, with reference to near-infrared fluorescent
Detection technique, can be quick, accurately detects specificity, solubility LP antigens in patient's sample.LP antigens near-infrared fluorescent is examined
Survey method high sensitivity, specificity is good, and sample collection and processing are simple in detection process, can fast and accurately obtain as a result, very
Accident scene and base is suitble to use.
Yellow fever antigen Near-infrared fluorescent detection technique high sensitivity, Idiotype is good, may be used in clinical detection, to face
Bed treatment provides qualitative foundation.
Specific embodiment
Bacteria Culture is the goldstandard of LP infection detections, but the condition of culture of LP is harsh, and time-consuming, is not suitable for clinical detection
And Site Detection;Because patient's Serum Antibody content can be only achieved detection level after infection LP the 3-6 weeks, disease early stage is not achieved
The requirement of diagnosis, therefore Serum Antibody Detection is usually used in the Retrospective epidemiological investigation of LP infection.LD patient is after infection 1-3 days
0 one polysaccharide antigens of LP with thermal stability and antitrypsic activity can be discharged by urine, the antigen is dense in urine
Degree is 30-100 times higher than in serum.Compared with EIA, the operation of near-infrared fluorescent detection method is easier, time-consuming shorter,
It is more preferably more promising detection means.
Yellow fever antigen near-infrared fluorescent detection method provided by the present invention, the laboratory result of appraisal show that the method is to LPl
The limit of identification of type antigen is l0ng/ml, cross reaction does not occur with other serotype antigens and various bacteria, is trained with bacterium
It is more consistent to support result, there is higher sensibility and specificity.Since yellow fever condition of culture requires harshness, the positive may be caused
Sample missing inspection.Generally speaking, the confidence level of near-infrared fluorescent method detection yellow fever and properties have reached clinical qualitative inspection
The requirement of survey is expected to be used for the quick early diagnosis of the legionaires' disease caused by yellow fever.
Illustrate embodiments of the present invention below by way of specific specific example, those skilled in the art can be by this specification
Disclosed content understands other advantages and effect of the present invention easily.The present invention can also pass through in addition different specific realities
The mode of applying is embodied or practiced, the various details in this specification can also be based on different viewpoints with application, without departing from
Various modifications or alterations are carried out under the spirit of the present invention.
Before further describing the specific embodiments of the present invention, it should be appreciated that protection scope of the present invention is not limited to down
State specific specific embodiment;It is also understood that the term used in the embodiment of the present invention is specific specific in order to describe
Embodiment, the protection domain being not intended to be limiting of the invention;In description of the invention and claims, unless in text
In addition explicitly point out singulative "one", " one " and " this " include plural form.
When embodiment provides numberical range, it should be appreciated that except non-present invention is otherwise noted, two ends of each numberical range
Any one numerical value can be selected between point and two endpoints.Unless otherwise defined, in the present invention all technologies for using and
Scientific terminology is identical with the normally understood meaning of those skilled in the art of the present technique.Except used in embodiment specific method, equipment,
Outside material, according to record of the those skilled in the art to the grasp of the prior art and the present invention, it can also use and this
Any method, equipment and the material of the similar or equivalent prior art of method, equipment described in inventive embodiments, material come real
The existing present invention.
Unless otherwise stated, disclosed in this invention experimental method, detection method, preparation method using this technology lead
Molecular biology, biochemistry, chromatin Structure and the analysis of domain routine, analytical chemistry, cell culture, recombinant DNA technology and
The routine techniques of related field.These technologies are existing in the prior art perfect to illustrate
Test strain is provided by Military Medical Science Institute in various embodiments of the present invention;Bacterium used in cross-over experiment is trained by this laboratory
It supports and preserves.Near-infrared fluorescent detector is developed by Kaichuang Biotechnology Co. Ltd., Shanghai and provided.
Embodiment 1
1. sputum is acquired and is preserved
In the case where medical worker instructs, firmly the sputum in expectoration respiratory tract deep, spits to sterile wide-mouth bottle patient, covers tightly immediately
Son saves backup, and pending sample should be placed in 4 DEG C of refrigerators and preserve.
2. urine capture and preservation
Patient cleans under the guidance of medical worker and collects first time in early morning urine, and the urine of collection is mounted in clean sterile appearance
It is saved backup in device.Urine specimen can preserve 72 hours under the conditions of 2-8 DEG C, then must be by sample if you need to preserve the longer time
Product are stored under -20 DEG C of environment.Multigelation sample should be avoided, otherwise can generate the experimental result of mistake.Sample can not preserve
In automatic defrosting refrigerator.
3. the preparation of antigen:
Yellow fever inoculation is bought on buffers active carbon yeast agar medium (BCYE) from ATCC,
37℃、5%C02Under conditions of cultivate, with physiological saline eluting bacterial from tablet after 3-5 days, 5000rpm centrifugations obtain
Bacterium mud (while removing the liquid component that tablet is eluted), bacterium mud add appropriate PBS buffer solution sonic disruption, and it is pure to cross anion column
Change, washed with 50mM NaCl, then eluted with 150mM NaCl, eluent dress bag filter embeds water suction concentration with PEG, obtains anti-
It is former.
4. the preparation of LP monoclonal antibodies:
Animal immune:Selection 6-8 week old BALE/C mouse are immunized, and antigen adds the Fu Shi of equivalent to help completely when immune for the first time
Agent is fully emulsified, and for the second time and third time antigen adds equivalent freund 's incomplete adjuvant fully emulsified, and 3 days abdominal cavities are straight before cell fusion
Injections of antigens booster immunization is connect, one exempts from antigen Freund Freund's complete adjuvant, and subcutaneous inoculation, amount of antigen is 0.lmg/ mouse, 2 after 15 days
Exempt from, subcutaneous inoculation antigen Freund Freund's incomplete adjuvant, amount of antigen is 0.2mg/ mouse, and three exempt from antigen Freund and not exclusively help after 25 days
Agent, subcutaneous inoculation, amount of antigen are 0. 2mg/ mouse, tail are cut after 10 days, blood is taken to survey ELISA potency, choose potency and are more than 100,000 times
Mouse carry out booster immunization, aqua intraperitoneal injection antigen 1 mg/ mouse, spleen cell is taken to merge after three days;Cell fusion:Using
Eyeball excise depletion method puts to death mouse, and sterile working takes out spleen, prepares bone-marrow-derived lymphocyte suspension, by bone-marrow-derived lymphocyte with preparing
The good mixing of the homology SP2/0 myeloma cell in exponential phase, prepares hybridoma;Filtering hybridoma:With
HAT medium culture cells can filter out the hybridoma of myeloma cell and bone-marrow-derived lymphocyte after 2 weeks;Hybridoma
Cloning:The method of cloning is limiting dilution assay, is carried out according to the conventional method in laboratory, cloning 3-4 times, until gram
Until 100% antibody positive is detected in each hole of grand cell growth;The preparation of antibody:BALB/C mice is taken, stone is injected intraperitoneally first
Wax, after 1-2 weeks, intraperitoneal inoculation hybridoma adjusts hybridoma concentration to 1 × 105A/ml presses every mouse abdomen
Chamber injection lml inoculations.After 1-2 weeks, ascites is extracted with syringe, you can obtain a large amount of monoclonal antibody.Monoclonal antibody
Purifying:HiTraprProtein A FF 5m1 prepackage chromatography antibody is selected, the antibody for collecting purifying is spare.
5. immunofluorescence LP antibody particle preparations:
With 0.01mol/L, the phosphate buffer of PH7. 3 ± 0.05 centrifuges the fluorescent latex particles of a diameter of 150nm clearly
It washes 2 times and latex particle is dispersed in addition LP antibody in cleaned latex particle;Adding in EDCA makes antibody be coupled with particle;
Group extra on ethanol amine closing latex particle is added in by the antibody to be marked of every milligram of 100 microgram of microballoon latex, 100 micrograms
EDCA dosages label.Every milligram of microballoon latex is finally plus the ethanol amine of 20 micrograms is closed.
6. the preparation of kit:
LP antibody is diluted to optimum concentration 2.0mg/ml with the PBS buffer solution of 0.01M pH7.2, utilizes trace of albumin point membranous system
Solution spray is added on the nitrocellulose filter in appropriate aperture, carries out spray film by the amount of lul/cm, 37 DEG C of baking ovens are dried for standby.With
The one antibody complex sized glass fibres film of fluorescent latex particles of preparation, drying for standby in vacuum drier.Practise each coating
The nitrocellulose filter of LP antibody, the glass fibre element film for being marked with immune fluorescent probe, sample pad, blotting paper and backboard assembling
It is prepared into yellow fever antigen near-infrared fluorescent detection kit.
Embodiment 2
Yellow fever antigen near-infrared fluorescent detection method and sputum culture contrast experiment:
Sputum culture:Sputum is pre-processed into (the side combined using 1ug/ml trypsin solutions with glass fragment oscillating phase
Method inoculates culture after carrying out pre-treatment to sputum), pretreated sample is seeded in immediately on GVPC selective mediums, right
The bacterium colony of doubtful yellow fever carries out Grain stain, and by the doubtful colony inoculation of Grain stain feminine gender in BCYE-a and BCYE-Cys
On agar plate, 37 DEG C of incubator culture 2d.Every grown on BCYE-a culture mediums and do not grown on BCYE-Cys culture mediums
Bacterium colony i.e. be regarded as yellow fever.
Kit detects:
Sample to be tested is sputum dilution, is diluted using the PBS buffer solution of 0.01M pH7.2, dilution ratio 1:l.Treat test sample
Product and kit, which are balanced to room temperature, to be started to detect, and 3 drop sample (about 120- are added dropwise in the well of each kit with dropper
150u1).At 15 minutes, fluorescence signal is detected, and judged with fluorescent instrument with near-infrared fluorescent detector, analyzer
Detection range to fluorescence signal is AD value 0-10000, and according to the performance of instrument, CUTOFF values are 50, and detection AD values are more than etc.
It is positive findings in 50.Experimental result and bacteria cultivation results contrast verification.For the sputum sample of Bacteria Culture and urine sample
This is from identical patient.The results are shown in Table 1 with sputum culture for yellow fever antigen near-infrared fluorescent detection method:
Table 1
Sensitivity=75%
Specificity=94. 11%
In addition, during using urine as sample to be tested, testing result is consistent with sputum culture testing result.
It can be seen from the table 2 in the table 1 and embodiment 3 in embodiment 2 in the detection of totally 30 parts of clinical samples, warp
The near-infrared fluorescent detection method detection LP positives are 4, and feminine gender is 26;Sputum culture LP positive samples are 3, negative sample
This is 27.The sensitivity of this method is 75%, and specificity is 94.11%;Cross reaction does not occur with other bacteriums;It is minimum to measure
It measures as l0ng/ml, stability and repeatability are good.
In conclusion the present invention effectively overcomes various shortcoming of the prior art and has high industrial utilization.
The above-described embodiments merely illustrate the principles and effects of the present invention, and is not intended to limit the present invention.It is any ripe
The personage for knowing this technology all can carry out modifications and changes under the spirit and scope without prejudice to the present invention to above-described embodiment.Cause
This, those of ordinary skill in the art is complete without departing from disclosed spirit and institute under technological thought such as
Into all equivalent modifications or change, should by the present invention claim be covered.
Claims (8)
1. a kind of yellow fever antigen near-infrared fluorescent detection kit, including the reagent strip being located on backboard, reagent strip is from sample-adding
End starts to be followed successively by sample pad, is marked with the glass fibre element film of immune fluorescent probe, is coated with the nitrocellulose of LP antibody
Film and blotting paper.
2. a kind of yellow fever antigen near-infrared fluorescent detection kit as described in claim 1, which is characterized in that described immune
Fluorescence probe is the coated near-infrared fluorescent latex beads particle of LP antibody.
3. the preparation side of the yellow fever antigen near-infrared fluorescent detection kit as described in claim 1-2 any claims
Method includes the following steps:
1) by fluorescent latex particles eccentric cleaning and latex particle is disperseed with phosphate buffer, in cleaned latex particle
Middle addition LP antibody;Adding in EDCA makes antibody be coupled with particle;Add group extra on ethanol amine closing latex particle, system
It is standby to obtain probe solution;
2) it will be added on nitrocellulose filter containing the spray of the buffer solution of LP antibody using trace of albumin point membranous system, drying is standby
With;
3) with the probe solution sized glass fibres film for preparing, drying for standby in vacuum drier;
4) by the coating nitrocellulose filter of LP antibody, the glass fibre element film for being marked with immune fluorescent probe, sample pad, water suction
Paper and backboard assembling are prepared into yellow fever antigen near-infrared fluorescent detection kit.
4. the preparation method of yellow fever antigen near-infrared fluorescent detection kit as claimed in claim 3, which is characterized in that institute
It states in step 1, a diameter of 140-160nm of fluorescent latex particles.
5. the preparation method of yellow fever antigen near-infrared fluorescent detection kit as claimed in claim 3, which is characterized in that institute
State in step 1, latex particle, LP antibody, EDCA weight ratio be 9-11:0.9-1.1:0.9-1.10.
6. the preparation method of yellow fever antigen near-infrared fluorescent detection kit as claimed in claim 3, which is characterized in that institute
It states in step 2, the spray dosage of LP antibody is:The antibody-solutions of 1.8-2.2mg/ml are prepared immune by the discharge rate of 0.9-1.l μ L/cm
Nitrocellulose filter.
7. the preparation method of yellow fever antigen near-infrared fluorescent detection kit as claimed in claim 3, which is characterized in that institute
The preparation method for stating LP antibody is as follows:
1) preparation of antigen:By yellow fever inoculation on culture medium, physiological saline eluting bacterial from tablet is used after culture,
Centrifugation obtains bacterium mud, and bacterium mud adds appropriate amount of buffer solution sonic disruption, crosses anion column purifying, is eluted after washing, eluent dress dialysis
Bag embeds water suction concentration with PEG, obtains antigen;
2) preparation of monoclonal antibody:
Animal immune:Mouse is immunized;Cell fusion:Mouse is put to death, sterile working takes out spleen, prepares bone-marrow-derived lymphocyte
Suspension mixes bone-marrow-derived lymphocyte with the ready homology SP2/0 myeloma cell in exponential phase, prepares hybridization
Oncocyte, and carry out filtering hybridoma;The cloning of hybridoma:Each hole detection 100% of clone cell growth is anti-
Until the body positive;The preparation of antibody:Paraffin is injected intraperitoneally in mouse first, hybridoma, 1-2 are inoculated in 1-2 weeks pneumoretroperitoneum
Zhou Hou extracts ascites with syringe, can obtain a large amount of monoclonal antibody after purification.
8. the yellow fever antigen near-infrared fluorescent detection kit as described in claim 1-2 any claims resists in yellow fever
The purposes of former detection field.
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| CN201611163230.5A CN108226507A (en) | 2016-12-15 | 2016-12-15 | A kind of yellow fever antigen near-infrared fluorescent detection kit and application thereof |
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| CN201611163230.5A CN108226507A (en) | 2016-12-15 | 2016-12-15 | A kind of yellow fever antigen near-infrared fluorescent detection kit and application thereof |
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| CN201611163230.5A Pending CN108226507A (en) | 2016-12-15 | 2016-12-15 | A kind of yellow fever antigen near-infrared fluorescent detection kit and application thereof |
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Cited By (2)
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| CN108458996A (en) * | 2017-02-20 | 2018-08-28 | 镇江华维检测技术有限公司 | Yellow fever antigen near-infrared fluorescent detection kit |
| CN114200130A (en) * | 2021-12-10 | 2022-03-18 | 江苏硕世生物科技股份有限公司 | Preparation method, detection system and detection method of fluorescence detection test paper |
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| CN108458996A (en) * | 2017-02-20 | 2018-08-28 | 镇江华维检测技术有限公司 | Yellow fever antigen near-infrared fluorescent detection kit |
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