CN1834652A - Immunity chromatography test paper for detecting Brucella and its prepn. method - Google Patents
Immunity chromatography test paper for detecting Brucella and its prepn. method Download PDFInfo
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- CN1834652A CN1834652A CN 200610076701 CN200610076701A CN1834652A CN 1834652 A CN1834652 A CN 1834652A CN 200610076701 CN200610076701 CN 200610076701 CN 200610076701 A CN200610076701 A CN 200610076701A CN 1834652 A CN1834652 A CN 1834652A
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Abstract
This invention discloses immune chromatography test paper and its preparation method that used to detect bulushi germ antigen. The paper includes sample mat 1, golden mark mat that nearly connected to one end of the mat 1 and contains bulushi germ antibody mark colloidal gold probe 2, nitrate cellulose film 3 that connected to another end of the gold mat and water absorbing mat 4 connected to another end of the NC film, the NC film is coated by separated detection line 5 and quality control line 6, the detecting line is bulushi germ antibody, the quality control line is goat anti rabbit IgG. This invention is used to the antibody detecting of clinical specimen, pollution and environmental bulushi germ, also can be used to identification of pure culture bulushi germ.
Description
Technical field
The present invention relates to the brucellar test paper of a kind of detection, the brucellar immune chromatography test paper of particularly a kind of detection also relates to its preparation method.
Background technology
Cloth Lu Shi disease is a kind of infectious disease of infecting both domestic animals and human, and it is distributed in all over the world widely.Found that at present Brucella (Brucella) has 6 kinds, comprise Bacterium melitense (Br.Militensis), Bacillus abortus (Br.abotus), Brucella suis (Br.suis), Brucella melitensis (Br.ovis), wooden mouse brucella (Br.neotomae) and dog brucella (Br.canis), wherein Bacterium melitense, Bacillus abortus, Brucella suis and Brucella melitensis are caused a disease to the people.Brucella (Brucella spp) is Gram-negative brevibacterium or coccobacillus, the nutritional requirement harshness, and poor growth, first separation and Culture needed 7 days even the longer time just can grow the visible bacterium colony of naked eyes.Boivin antigen is brucellar surface antigen, also is brucellar common antigen not of the same race in the Brucella.
Nineteen forty-three the U.S.'s offensive biological warfare agent that begins one's study, and in the fight production of agent of Camp Detrick, the kind of the biological warfare agent of studying first and producing comprises that bacillus anthracis, plague bacillus, soil draw hot bacterium, Brucella suis, comma bacillus, Salmonella typhi, the hot Richettsia of Q, Venezuelan equine encephalitis virus, botulinum toxin and Type B staphylococcal enterotoxin etc.The biological warfare agent of the U.S.'s first weaponization in newly-built Pine Bluff munitions factory in 1954 promptly is a brucella.Up to now, brucella still as classical biological warfare agent, is put into " international Biological Weapons Convention " biological warfare agent inventory.After " 9.11 " terrorist incident, brucella is listed in the strong pathogenic microorganism that can cause bio-terrorism again in American National anti-terrorism prediction scheme.
Brucella can pass through the gasoloid hyperinfection, and 10～100 hyphopodiums are so that people's morbidity.Therefore, brucella fast detecting Research on New is the needs that biological defence and anti-bio-terrorism attack.
Because the brucella poor growth, in the biohazard accident that reply is caused by brucella, it is particularly important that the fast detecting of pathogen seems.Brucellar fast detecting mainly comprises the detection of specific antigen and the detection of specific nucleic acid fragment, what Detection of antigen was commonly used is immunofluorescent test and enzyme linked immunosorbent assay (ELISA), that detection of nucleic acids is commonly used is polymerase chain reaction (PCR), the use of these methods has many bibliographical informations [Hamdy ME, Amin AS.Detection of Brucella species in the milk of1nfected cattle, sheep, goats and camels by PCR.Vet J 2002,163 (3): 299-305; Morata P, Queipo-Ortuno MI.Development and evaluationof a PCR-enzyme-linked immunosorbent assay for diagnosis of humanbrucellosis.J Clin Microbiol 2003,41 (1): 144-148].
But in practice, the use of these methods exposes some common inferior positions, and for example: experiment must be carried out in special laboratory; Need power supply and special instrument; Operating personnel must pass through special training; Reagent needs refrigeration; Experimental system is not easy standardization, for making credible result, need carry out repeatedly trial test and set up multinomial contrast; The test operation complex steps needs more than 2 hours at least.Therefore, biohazard accident field condition and the use of methods such as immunofluorescent test, ELISA and PCR is restricted to the requirement of time.
Up to the present, do not see immune colloidal gold technique as yet and be used to detect brucellar report.
Summary of the invention
The object of the invention provides the brucellar immune chromatography test paper of a kind of detection (Immuno-Chromatographic Assay, ICA test paper).
The brucellar immune chromatography test paper of detection provided by the present invention (ICA test paper), comprise sample pad, closely be connected in the gold mark pad that contains Brucella antibody mark colloidal gold probe of described sample pad one end, with the close-connected nitrocellulose membrane of the other end (NC film) of described gold mark pad with closely be connected in the adsorptive pads of the described nitrocellulose membrane other end; Described nitrocellulose membrane is coated with detection line and the nature controlling line that is separated from each other, and described detection line is anti-brucella specific antibody, is preferably rabbit antibody, and described nature controlling line is anti-rabbit antiantibody.
Convenient in order to use, the following backboard that also closely is connected with of described adsorptive pads.The material of backboard can be diversified, as plastics, resin etc.
Second purpose of the present invention provides a kind of method for preparing the brucellar immune chromatography test paper of above-mentioned detection.
The method of the brucellar immune chromatography test paper of the above-mentioned detection of preparation provided by the present invention may further comprise the steps:
1) the brucella specific antibody can prepare as follows: subcutaneous multi-point injection immunity Fu Shi Freund's complete adjuvant brucella thalline.Every injection 2 * 10
8Cfu bacterium/ml injects 3 times altogether.Slide agglutination test detects serum titer and reaches above blood sampling in 1: 1280.
The brucella specific antibody is sprayed onto on the tunica fibrosa, and bag is obtained detection line by a zone of NC film; The antiantibody solution of anti-rabbit antiantibody is sprayed onto on the tunica fibrosa, and bag is obtained nature controlling line by another zone of NC film; 37 ℃ dry 2.5-3.5 hour, then the one end is sticked on the suction paper washer on;
2) preparation contains the immune colloid gold probe solution of brucella specific antibody mark; Get 5～5.5ml, adding 0.5～0.55g sucrose fully dissolves, glass fibre or polyester film are immersed this immune colloid gold probe solution, placed 10-12 hour for-20 ℃～-50 ℃, freeze dryer is drained and is promptly obtained gold mark pad, and it is sticked on an end of the close described detection line of the tunica fibrosa that step 1) obtains;
3) in step 2) in gold mark pad above paste sample pad again, obtain detecting brucellar immune chromatography test paper.
Convenient in order to use, the following also adhesive back of described adsorptive pads.
In brucellar immune chromatography test paper of detection provided by the present invention and preparation method thereof, described anti-brucella specific antibody mark colloidal gold probe can be prepared by following method:
1) with HAuCl
4Be mixed with 0.01% aqueous solution, get 100ml and be heated to and boil, stir the 1% trisodium citrate (Na that accurately adds 1.6ml down
3C
6H
5O
72H
2O) aqueous solution continued heated and boiled 10-12 minute, stablized into the grape wine redness up to liquid color, obtained colloidal gold solution, and cooling back water returns to original volume;
2) use K
2CO
3Or the HCl adjust pH is 8.5-9.5, adds the brucella specific antibody by 30 μ g/ml, stirs 25-35 minute, prepares the immune colloid gold probe solution of brucella specific antibody.Add 10%BSA 5ml then, stirred 20-30 minute, add 1ml 10%PEG20000, stirred 20-30 minute, 20,000-23, the centrifugal 25-35 of 500g minute, abandon supernatant, collecting precipitation is preserved liquid with sodium tetraborate and is collected, and obtains collaurum mark probe.
The K of described adjusting pH value
2CO
3Concentration can be 0.15-0.25M, be preferably 0.2M; The concentration of described adjusting pH value HCl can be 0.08-0.12mol/L, is preferably 0.1mol/L.
Immune colloidal gold technique detects brucellar ultimate principle: with antibody sandwich cellulose nitrate (NC) film of anti-brucella surface antigen, in order to catch brucella or the brucellergen in the sample, the immune colloid gold probe in detecting of specific antibody of having used mark then.Brucella in the sample or brucellergen macroscopic precipitation line promptly occurs after analysing through about 5 minutes ply of paper.This technology and other method relatively, advantage is: sample disposal is simple in the testing process, does not need specialized equipment and staff training, non-specialized-technical personnel can operate to specifications, and observations rapidly, are well suited for the on-the-spot and basic unit's use of accident.
Immune chromatography test paper of the present invention adopts double antibody sandwich method, to resist the brucella specific antibody to be coated on the nitrocellulose filter, be used for catching the brucellergen of sample, the immune colloid gold probe with cloth Lu Shi specific antibody mark detects then.
Test paper of the present invention can be used for brucellar detecting in clinical samples, pollutant and the environment, also can be used for the brucellar evaluation of pure culture.Advantage of the present invention is that sample disposal is simple in the testing process, does not need specialized equipment and staff training, and non-specialized-technical personnel can operate to specifications, and observations rapidly, is well suited for the on-the-spot and basic unit's use of accident.
Description of drawings
Fig. 1 is the structural representation of brucella immune chromatography test paper.Immune chromatography test paper is made up of adsorptive pads 4, nitrocellulose membrane 3, gold mark pad 2 and glass fibre membrane sample pad 1 four parts; Be coated with detection line 5 and nature controlling line 6 on the nitrocellulose membrane 3.
Embodiment
The preparation of embodiment one, the brucellar immune chromatography test paper of detection
1 main material
Gold chloride (HAuCl
4), available from Sigma company, 1g/ bottle packing.
Nitrocellulose membrane (NC film), sample pad and absorbent filter are available from Millipore company.
Plastic back plate, Beijing Yan Hua company.
Miscarriage (ox) brucella strain (preserving number 210105) is drawn from Beijing biological products assay institute existing by the Micro biological Tests research centre strain library preservation of the entire PLA of Military Medical Science Institute.
2 methods
Experimental technique among the following embodiment if no special instructions, is conventional method.
Percentage composition among the following embodiment if no special instructions, is the quality percentage composition.
2.1 the preparation of brucella specific antibody
2.1.1 the preparation of brucellergen
Miscarriage (ox) brucella (Microbiology and Epidemic Disease Inst., Academy of Military-Medical Sciences's strain library is protected a surname, preserving number 210105) is seeded on pancreatin soybean blood agar (the 5% sheep blood) flat board 37 ℃ of 5%CO
2Incubator is cultivated, and observes dull and stereotyped colonial morphology, the typical single bacterium colony of picking, transferred species pancreatin soybean blood agar inclined-plane, 37 ℃ of 5%CO of going up
2Cultivated 48-50 hour in the incubator.The inclined-plane of cultivating is washed lawn with stroke-physiological saline solution, collect bacterium liquid, boiling water bath 10-15 minute, be cooled to room temperature, transferring bacteria concentration with stroke-physiological saline solution is 2 * 10
8Cfu bacterium/ml is the brucella somatic antigen, 4 ℃ of preservations.
2.1.2 the preparation of brucella specific antibody
Select body weight 2kg Healthy female white big ear rabbit (available from Academy of Military Medicine, PLA's animal center) for use, subcutaneous multi-point injection immunity Fu Shi Freund's complete adjuvant brucella somatic antigen 2 * 10
8Cfu bacterium/1ml/ only, respectively at the 20th day behind the initial immunity, the 30th day, the 40th supplementary immunization aqua 1 pin, boost and approach be with identical for the first time, last immunity examination in back 10 days blood, slide agglutination test detects serum titer and reaches 1: 1280 above blood sampling.
Adopt conventional saturated ammonium sulfate salting out method, 33% saturated ammonium sulfate is saltoutd 2 times, collects antibody behind the dialysis desalting.
3 preparation immune colloid gold probes
3.1 preparation colloidal gold solution: adopt the citrate reducing process to prepare colloid gold particle, concrete grammar is: with HAuCl
4Be mixed with 0.01% aqueous solution, get 100ml and be heated to boiling, stir the 1% trisodium citrate (Na that accurately adds 1.6ml down
3C
6H
5O
72H
2O) aqueous solution, liquid color are stablized into the grape wine redness, promptly obtain colloidal gold solution.The cooling back returns to original volume with distilled water, makes the colloid gold particle that particle diameter is 25nm.
3.2 determine collaurum coupling antibody saturation concentration: use 0.2M K
2CO
3Or 0.1M HCl adjusting colloidal gold solution pH8.5-9.5, prepare 5 clean tube, add the 1ml colloidal gold solution respectively.Purified anti-brucella specific antibody dilution is 1mg/ml, adds 10 μ l, 15 μ l, 25 μ l, 35 μ l respectively in 4 test tubes, another is contrast, under room temperature, placed 5 minutes behind the mixing, add the 10%NaCl aqueous solution, mixing leaves standstill after 10-20 minute and observes liquid color.Contained minimum antibody amount was the optimum concentration of stablizing the required antibody of 1ml collaurum when the colloidal gold solution color was constant, increased by 20% antibody amount based on this, was collaurum coupling antibody saturation concentration.The result shows: keeping the constant antibody amount of colloidal gold solution color is 25 μ l, i.e. 25 μ g/ml, and selecting the coupling antibody concentration is 30 μ g/ml.
3.3 the preparation of gold mark pad 2: preparation contains the immune colloid gold probe solution 50ml that concentration is the anti-brucella specific antibody of 30 μ g/mL as stated above, stirred 25-35 minute, add 10%BSA 5ml, stirred 20-30 minute, add 1ml 10%PEG20000, stirred 20-30 minute, 20,000～23, the centrifugal 25-30 of 500g minute, abandon supernatant, precipitation is once preserved liquid collecting precipitation 5ml with sodium tetraborate in the back with the sodium tetraborate washing.Get gold mark probe 5ml and add 0.5g sucrose, fully dissolving evenly is added on the glass fibre membrane, places 10-12 hour for-20 ℃～-50 ℃, and freeze dryer is drained, and obtains gold mark pad 2.
4, the preparation of brucella quick detection test paper
As shown in Figure 1, detecting brucellar immune chromatography test paper is made up of adsorptive pads 4, nitrocellulose membrane 3, gold mark pad 2 and glass fibre membrane sample pad 1 four parts; Be coated with detection line 5 and nature controlling line 6 on the nitrocellulose membrane 3.
4.1NC the bag quilt of film 3:
With 0.01M pH7.2PB damping fluid (NaH
2PO
4.2H
2O 0.39g, Na
2HPO
41.07g, deionized water 1000ml) and the anti-brucella specific antibody of dilution, concentration is 4mg/ml, is used to wrap tested survey line.PBS (NaH with 0.01M pH 7.2
2PO
4.2H
2O 0.39g, Na
2HPO
41.07g, NaCl 8.5g, deionized water 1000ml) and the anti-rabbit antiantibody of dilution, concentration is 4mg/ml, is used for bag by nature controlling line.Be sprayed on respectively on the thick nitrocellulose filter of 2.5mm with the XYZ3000 of BIODOT company Membrane jetter, form detection line 5 and 6,37 ℃ of dry 2.5-3.5h of nature controlling line of being separated from each other.
4.2 the preparation of brucella quick detection test paper
5 detect the preparation of brucellar immune chromatography reagent kit
Use for convenience, with the following plastic back plate that closely connects again of the immune chromatography test paper of the brucella fast detecting of step 3 preparation, the test paper that again this is had a backboard is packed in the kit, add drying agent after sealing preserve.This kit is provided with the point sample mouth corresponding to the position of sample pad, is provided with observation window corresponding to the position that contrasts band and calibration tape.
The using method of 6 brucella quick detection test paper and principle
During mensuration test strips sample pad 1 is immersed in the liquid sample, sample pad 1 is that imbitition moves to the upper end, and the gold mark pad of flowing through made the immune Au composite on the dry plate redissolve at 2 o'clock, and drives it and ooze to nitrocellulose membrane 3 and move.If specific antigen to be measured is arranged in the sample, its can with the antibodies of immune Au composite, this antigen antibody complex flow to detection line 5 and is promptly obtained by insolubilized antibody, shows red reaction lines on film.Superfluous immune Au composite continues to move ahead, and combines to the anti-rabbit antiantibody of nature controlling line 6 and solid phase, and shows red Quality Control lines.Otherwise, the then reactionless lines of negative sample, and only show the Quality Control lines.
Embodiment two, brucellar detection reach the cross matching with other Related Bacteria
1 brucellar detection
1.1 by Microbiology and Epidemic Disease Inst., Academy of Military-Medical Sciences (C Micro biological Tests research centre provide, concentration is 1 * 10
6The 3 strain brucella of cfu/ml comprise Brucella abortus 210105 strains, Brucella melitensis 210301 strains and Brucella suis 210401 strains, and bacteria suspension is standby as sample detection liquid.
1.2 the bag through embodiment one preparation is positive findings by the detection of the immune chromatography test paper of brucella specific antibody and anti-rabbit antiantibody.
The cross matching of 2 other Related Bacteria
2.1 by Microbiology and Epidemic Disease Inst., Academy of Military-Medical Sciences (C Micro biological Tests research centre provide, concentration is 1 * 10
8The soil of cfu/ml draws hot bacterium 410062 strains and plague bacillus 410050 strains, and bacteria suspension is standby as sample detection liquid.
2.2 the bag through embodiment 1 preparation is negative findings by the detection of the immune chromatography test paper of brucella specific antibody and anti-rabbit antiantibody.
Embodiment three, laboratory examination
1 specimen preparation
Experiment uses bacterium 44 strains, wherein brucella 33 strains, plague bacillus, soil to draw each 1 strain of hot bacterium, pseudonodule Yale Salmonella, enterocolitis Yale Salmonella, legionella pneumophilia, colon bacillus, shigella dysenteriae, salmonella, proteus, Pseudomonas aeruginosa and comma bacillus altogether.33 strain brucella comprise Brucella abortus 15 strains, Brucella melitensis 12 strains, Brucella suis 6 strains.Above-mentioned bacterium is inoculated suitable culture base separately respectively, and under different conditions, cultivate, wash with stroke-physiological saline solution after growing lawn, turbidimetry preparation brucella bacteria suspension 1 * 10
6Cfu/ml, 1 * 10
7Cfu/ml and 1 * 10
8Cfu/ml, other bacterium bacteria suspension 1 * 10
8Cfu/ml, liquid is standby as detecting.
2 experimental techniques
Get the brucellergen quick detection reagent, respectively at adding 3 of above-mentioned sample detection liquid (about 150 μ l) on the sample pad, begin observations after 2 minutes, observation in 15 minutes stops.Result's report: it is negative that 1 precipitation line appears in the nature controlling line place, promptly do not have brucellergen and detect; It is positive that 2 precipitation lines appear in detection line and nature controlling line place, promptly has brucellergen to detect.
3 experimental results
The brucellergen quick detection reagent laboratory result of appraisal see Table 1.
The examination of table 1 brucellergen quick detection reagent (colloidal gold method) laboratory
| Bacteria name | The strain number | Bacterium amount (cfu/ml) | ||
| 10 6 | 10 7 | 10 8 | ||
| Brucella abortus | 15 | + | + | + |
| Brucella melitensis | 12 | + | + | + |
| | 6 | + | + | + |
| Plague bacillus | 1 | - | ||
| Soil draws hot bacterium | 1 | - | ||
| Pseudonodule Yale Salmonella | 1 | - | ||
| Enterocolitis Yale Salmonella | 1 | - | ||
| Legionella pneumophilia | 1 | - | ||
| Colon bacillus | 1 | - | ||
| Shigella dysenteriae | 1 | - | ||
| Salmonella | 1 | - | ||
| Proteus | 1 | - | ||
| Pseudomonas aeruginosa | 1 | - | ||
| Comma bacillus | 1 | - | ||
As can be seen from Table 1, the brucellar immune chromatography test paper of a kind of detection can detect 1 * 10
6Cfu/ml ox, sheep and Brucella suis be not with 1 * 10
8Cfu/ml plague bacillus, soil draw hot bacterium, artificial tuberculosis yersinia genus, yersinia enterocolitica, legionella pneumophilia, colon bacillus, shigella dysenteriae, salmonella, Pseudomonas aeruginosa and comma bacillus that cross reaction takes place.
Claims (7)
1, the brucellar immune chromatography test paper of a kind of detection, comprise sample pad (1), closely be connected in the gold mark pad (2) that contains brucella specific antibody mark colloidal gold probe of described sample pad one end, with the close-connected nitrocellulose membrane of the other end (3) of described gold mark pad with closely be connected in the adsorptive pads (4) of the described nitrocellulose membrane other end; Described nitrocellulose membrane is coated with detection line (5) and the nature controlling line (6) that is separated from each other, and described detection line is the brucella specific antibody, and described nature controlling line is anti-rabbit antiantibody.
2, the brucellar immune chromatography test paper of detection according to claim 1, it is characterized in that: the brucella specific antibody is preferably rabbit antibody.
3, the brucellar immune chromatography test paper of detection according to claim 1 is characterized in that: described nature controlling line is anti-rabbit antiantibody.
4, the brucellar immune chromatography test paper of detection according to claim 1 is characterized in that: described sample pad, adsorptive pads, gold mark pad are made by absorbent material; The following backboard that also closely is connected with of described adsorptive pads.
5, the brucellar immune chromatography test paper of detection according to claim 1 and 2, it is characterized in that: described brucella specific antibody mark colloidal gold probe is prepared by following method: 5, a kind of method that detects brucellar immune chromatography test paper for preparing may further comprise the steps:
1) preparation brucella specific antibody is sprayed onto the brucella specific antibody on the tunica fibrosa, and bag is obtained detection line by a zone of NC film; The antiantibody solution of anti-rabbit igg is sprayed onto on the tunica fibrosa, and bag is obtained nature controlling line by another zone of NC film; 37 ℃ dry 2.5-3.5 hour, then the one end is sticked on the suction paper washer on;
2) preparation collaurum mark probe is with HAuCl
4Be mixed with 0.01% aqueous solution, get 100ml and be heated to and boil, stir 1% trisodium citrate aqueous solution that adds 1.6ml down, continued heated and boiled 10-12 minute, cooling back water returns to original volume, uses K
2CO
3Adjust pH is 8.5-9.5, presses 30ug/ml and adds the brucella specific antibody, stirs 25-35 minute, add 10%BSA5ml then, stirred 20-30 minute, and added 1ml 10%PEG20000, stirred 20-30 minute, 20,000g-23, the centrifugal 25-35 of 500g minute, the sucking-off supernatant, precipitation is preserved liquid with sodium tetraborate and is collected, and obtains collaurum mark probe.
3) preparation contains brucella specific antibody labelled immune colloidal gold probe solution; Get 5-5.5ml, adding 0.5-0.55g sucrose fully dissolves, glass fibre or polyester film are immersed this immune colloid gold probe solution, placed 10-12 hour for-20 ℃～-50 ℃, freeze dryer is drained and is promptly obtained gold mark pad, and it is sticked on an end of the close described detection line of the tunica fibrosa that step 1) obtains;
4) in step 2) in gold mark pad above paste sample pad again, obtain detecting brucellar immune chromatography test paper.
6, method according to claim 5 is characterized in that: the concentration of described brucella specific antibody is 4mg/ml; The concentration of anti-rabbit antiantibody is 4mg/ml; The following backboard that also is pasted with of described adsorptive pads; The K of described adjusting pH value
2CO
3Concentration be 0.15-0.25M.
7, the kit that contains the brucellar immune chromatography test paper of arbitrary described detection among the claim 1-4.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101055274B (en) * | 2007-06-08 | 2012-07-11 | 中国人民解放军军事医学科学院军事兽医研究所 | Animal brucella antigen colloidal gold test paper film detection reagent kit |
| CN101055273B (en) * | 2007-06-08 | 2013-01-09 | 中国人民解放军军事医学科学院军事兽医研究所 | Animal brucella antibody colloidal gold test paper film detection reagent kit |
| CN101539574B (en) * | 2009-04-29 | 2013-01-16 | 北京市肿瘤防治研究所 | Application of Survivin antibody and esophageal cancer immunochromatography detecting test strip prepared therewith |
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| CN111879925A (en) * | 2019-12-30 | 2020-11-03 | 杭州奥泰生物技术股份有限公司 | Colloidal gold immunochromatographic assay test paper for rapidly diagnosing brucellosis |
| CN114075551A (en) * | 2021-06-11 | 2022-02-22 | 华中农业大学 | Monoclonal antibody of brucella salina lipopolysaccharide and application thereof |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101055274B (en) * | 2007-06-08 | 2012-07-11 | 中国人民解放军军事医学科学院军事兽医研究所 | Animal brucella antigen colloidal gold test paper film detection reagent kit |
| CN101055273B (en) * | 2007-06-08 | 2013-01-09 | 中国人民解放军军事医学科学院军事兽医研究所 | Animal brucella antibody colloidal gold test paper film detection reagent kit |
| CN101539574B (en) * | 2009-04-29 | 2013-01-16 | 北京市肿瘤防治研究所 | Application of Survivin antibody and esophageal cancer immunochromatography detecting test strip prepared therewith |
| CN109655614A (en) * | 2019-02-26 | 2019-04-19 | 广州维佰生物科技有限公司 | Ox brucellosis virus fluorescence micro-ball immune chromatography test paper and preparation method thereof and detection method |
| CN111879925A (en) * | 2019-12-30 | 2020-11-03 | 杭州奥泰生物技术股份有限公司 | Colloidal gold immunochromatographic assay test paper for rapidly diagnosing brucellosis |
| CN114075551A (en) * | 2021-06-11 | 2022-02-22 | 华中农业大学 | Monoclonal antibody of brucella salina lipopolysaccharide and application thereof |
| CN114075551B (en) * | 2021-06-11 | 2024-01-26 | 华中农业大学 | Monoclonal antibody of brucella lipopolysaccharide of sarin mouse species and application |
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