CN102707053A - Carbohydrate chip for quickly diagnosing tuberculosis, preparation method thereof and tuberculosis diagnostic kit adopting carbohydrate chip - Google Patents
Carbohydrate chip for quickly diagnosing tuberculosis, preparation method thereof and tuberculosis diagnostic kit adopting carbohydrate chip Download PDFInfo
- Publication number
- CN102707053A CN102707053A CN2012101854397A CN201210185439A CN102707053A CN 102707053 A CN102707053 A CN 102707053A CN 2012101854397 A CN2012101854397 A CN 2012101854397A CN 201210185439 A CN201210185439 A CN 201210185439A CN 102707053 A CN102707053 A CN 102707053A
- Authority
- CN
- China
- Prior art keywords
- phase carrier
- solid phase
- carbohydrate chip
- tuberculosis
- chip
- 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.)
- Pending
Links
Images
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention relates to a carbohydrate chip for quickly diagnosing tuberculosis, which comprises a solid-phase carrier. The carbohydrate chip is characterized by further comprising a mycobacterium tuberculosis lipopolysaccharides antigen probe which is fixed on the solid-phase carrier through oximation reaction. The invention also relates to a preparation method of the carbohydrate chip and a tuberculosis diagnostic kit adopting the carbohydrate chip. The preparation method comprises the following steps: separating, extracting and purifying lipopolysaccharides in a mycobacterium tuberculosis cell wall; fixing LAM (lipoarabinomannan) and LM (lipomannan) probes in the solid-phase carrier, such as a nitrocellulose membrane or porous plate, and the like, by utilizing a method of oximation reaction, thereby obtaining the carbohydrate chip; and judging if the mycobacterium tuberculosis is infected, by bonding the carbohydrate chip with a corresponding antibody in blood serum and performing enzyme linked color-developing reaction. A cell wall lipopolysaccharides antigen with species specificity adopted by the carbohydrate chip only can be combined with a specific tuberculosis antibody, so that the detecting specificity and sensitivity of the tuberculosis are greatly increased.
Description
Technical field
The present invention relates to biological medicine external diagnosis reagent field, more particularly, relate to a kind of carbohydrate chip that is applied to tuberculosis rapid diagnosis, its preparation method and adopt its reagent box for tuberculate diagnosis.
Background technology
China is high infection the lungy, high incidence country, and number of patients lungy occupy second in countries in the world.Much's bacillus Clinical detection method has bacteriological detection, serology to detect and gene diagnosis at present.The bacteriological detection of tuberculosis patient is the possible means of diagnosis of tuberculosis, treatment and evaluating efficacy, and these means comprise BACTEC detection system, mycobacterium growth indication mechanism, bacteriophage reporting techniques.Even the detection method of these means is shortening the problem that still exists some not promote after this shortcoming detection time, like the expensive equipment of needs, standardized bacterium etc.Gene diagnosis is shorter to the time that pulmonary tuberculosis detection, Much's bacillus strain identification, drug resistance monitoring etc. have higher specificity and needs; But because of popularizing of its false-negative result appearred and effects limit such as the required instrument costliness of part laboratory facilities is arranged in its high specific on the contrary easily, these means mainly comprise Much's bacillus DNA cloning method, real-time fluorescence quantitative PCR technology, PCR-RFLP, dna probe technology and biochip technology.The inspection that Serological testing is convenient, simple to operate because of its sampling, fast, do not rely on pathogen is a clinical diagnosis important supplementary means lungy, but the specific immunity diagnosis depends on specific antigen.The humoral immunity dynamic rule is very complicated in the tuberculosis infection process, lack the theoretical foundation of antigen optimal combination, and traditional Serum Antibody Detection is because mark and detection technique are backward, so detection sensitivity is not high.
Present commercial combinatorial antibody detection kit is used the antibodies in specific protein antigen and the serum more, can't detect approach antibody in all tubercular's serum.These detection methods are the problem of aspect such as life period, specificity all, can not detect the entrained Much's bacillus of patient fast and accurately, has brought very big inconvenience for treatment lungy and control.Therefore, seeking fast and accurately, the new method of diagnosis of tuberculosis is a control important means lungy.
Carbohydrate chip is the new bio technology that after genetic chip and protein chip, is born, and has that the sample size that needs is few, advantages such as high flux and high specific, and its ultimate principle is similar with protein chip with genetic chip, all is based on the specific effect between the material.It is that the glycan molecule of a plurality of different structures is fixed on the matrix of chemical modification through effect covalently or non-covalently, and then protein, glycoprotein are waited for the alanysis method that the test sample article are tested, analyzed.Superiority is: one of which, biologically active is stable.The long preservation biologically active is constant at ambient temperature for carbohydrate chip.Its two, have wide range of applications.Be applied to the carbohydrate chip technology on the clinical medicine, can carry out quick diagnosis simultaneously multiple infectiousness and NCD.Because all cell surfaces all contain glucide, these carbohydrate molecules play an important role in the identifying of cellular elements.
Much's bacillus cell wall constituent fat AM (lipoarabinomannan with species specificity; LAM), fat mannan (lipomannan; LM) be immune response regulatory factor important in the tuberculosis infection process; Be the important part of Much's bacillus and macrophage and BMDC effect, it is the research target spot of tuberculosis pathogenesis, immunopathogenesis, diagnoses and treatment as the important antigen of Much's bacillus always.The foreign study person detects LAM-IgG and is used for diagnosis of tuberculosis and did research, and develops corresponding diagnostic kit.Domestic also have the scholar to carry out similar research, but be mostly that directly buying external antigen or kit carries out application evaluation.Dependence on import reagent, price are comparatively expensive, have limited this Application for Diagnosis Technology and have promoted.
The present invention is intended on the basis of carbohydrate chip technology of independent development; Utilize the method for the fixing sugared probe of oximation reaction; The boivin antigen that purifying is obtained is fixed on the solid phase carrier, and applies to serodiagnosis, sets up a kind of quick; Accurately based on the technological tuberculosis rapid diagnosis kit of carbohydrate chip, for the tuberculosis prevention and control provide effective means.
Summary of the invention
Technical matters to be solved by this invention is, provides a species specificity good, highly sensitive and be applicable to the tuberculosis method for quick based on two kinds of liopopolysaccharides antigens and antibody of extensive detection.
In order to address the above problem; One object of the present invention is to provide a kind of carbohydrate chip that is used for tuberculosis rapid diagnosis; It comprises solid phase carrier; It is characterized in that said carbohydrate chip also comprises Much's bacillus liopopolysaccharides antigen probe, said Much's bacillus liopopolysaccharides antigen probe is fixed on the said solid phase carrier through oximation reaction.
In one embodiment, said Much's bacillus lipopolysaccharides extracts from Much's bacillus H
37The Rv bacterial strain.
In one embodiment, said Much's bacillus lipopolysaccharides is lipopolysaccharides LAM and LM.
In one embodiment, said solid phase carrier is nitrocellulose membrane or the microwell plate through chemical modification.
In one embodiment, the chemical modification of said solid phase carrier is to carry out according to following steps:
(1) solid phase carrier is soaked into epoxy silane/toluene solution, form the monofilm of epoxide function;
(2) the above-mentioned solid phase carrier that obtains is used α, ω-diamines-polyglycol (M
W=2000) solution-treated makes the polyglycol individual layer molecule that covers one deck band primary amino radical on the solid phase carrier;
(3) place the phosphate buffer of tertbutyloxycarbonyl aminooxy group acetate, ethene and N-hydroxy-succinamide to handle amidized solid phase carrier, handle with HCl/ acetic acid the washing back, makes primary amino radical convert azanol to through chemical modification.
Another object of the present invention is to provide preparation the said carbohydrate chip method that is used for tuberculosis rapid diagnosis, may further comprise the steps:
(1) extraction, purifying Much's bacillus cell membrane liopopolysaccharides antigen;
(2) utilize the method for oximation reaction that the liopopolysaccharides antigen probe is fixed in the solid phase carrier through chemical modification, be made into carbohydrate chip.
Wherein, the chemical modification of said solid phase carrier is carried out through aforesaid step.
A purpose more of the present invention is to provide a kind of tuberculosis detection kit, and said tuberculosis detection kit comprises the above-mentioned carbohydrate chip that is used for the tuberculosis fast detecting.
This kit is based on liopopolysaccharides antigen in the Much's bacillus cell membrane---and (lipoarabinomannan, LAM) (lipomannan LM) is the Much's bacillus method for quick of target to the fat AM with the fat mannan.
The present invention provides a kind of Much's bacillus carbohydrate chip detection method based on lipopolysaccharides and antibody.The detection method that invention is provided is that the antigenicity substance lipopolysaccharides with bacterial classification, the specific Much's bacillus cell membrane of bacterial strain surface is made into chip through a series of processing; Utilize itself and the lipopolysaccharides and the specificity of antibody to combine to detect whether contain lipopolysaccharides and antibody in the blood serum sample; And this lipopolysaccharides and antibody identified; And then the bacterial strain kind of the entrained Much's bacillus of definite patient, for the treatment provider in later stage to.May further comprise the steps:
1, the extraction of Much's bacillus lipopolysaccharides and silver dye the colour developing evaluation
2, the purifying of lipopolysaccharides and evaluation
3, quantitative sugared constituent analysis
(1) anthrone method is measured the content of polysaccharide in the lipopolysaccharides extract
(2) discriminating has or not protein contamination:
4, the lipopolysaccharide molecule amount is identified
5, the antigenicity of elisa assay Much's bacillus CA
6, the making of carbohydrate chip
7, chip detection
The present invention has the following advantages:
(1) specificity is good: the lipopolysaccharides of Much's bacillus cell membrane has very high antigentic specificity, can be with lipopolysaccharide antibody resisting specific bond in the serum;
(2) highly sensitive: it is few to have test sample, characteristics such as flux height;
(3) cheap: as not need the equipment of expensive or special reagent, only need add testing sample during detection, for the user saves a large amount of financial resource and material resource;
(4) be applicable to extensive detection: this product method of application is simply quick, does not have specific equipment, reagent requirement, thus can use at any environment, can large-scale promotion.
(5) time is short: required time is no more than 2 hours.
The present invention uses this method and has designed a carbohydrate chip of on the sheet base, placing two kinds of liopopolysaccharidess that from Much's bacillus type strain H37Rv strain cell wall, extract respectively; The tuberculosis that can fast detecting causes by Much's bacillus; Make shorten detection time lungy and be no more than 2 hours, fully shown the superiority of this method.Therefore this method might develop into the new method that is applied to clinical fast detecting, for control lungy plays an important role.
Description of drawings
Fig. 1 is the carbohydrate chip mode chart; The negative Quality Control point of cloudy ginseng among the figure, the positive Quality Control point of sun ginseng, H37Rv-LM is the LM antigen of Much's bacillus type strain H37Rv; H37Rv-LAM is the LAM antigen of Much's bacillus type strain H37Rv, every kind of equal triplicate of antigen point sample.
Embodiment
Elaborate in the face of embodiment provided by the invention down.
Embodiment 1
1, the mycobacterium lipopolysaccharides mentions that slightly silver dyes colour developing and identifies
Cultivate and collect Much's bacillus type strain H
37Rv strain (bacterium type strain H
37The Rv strain is identified institute available from Chinese pharmaceutical biological product), 2 hours deactivation bacteriums of 100 ℃ of water-baths.Use chloroform: methyl alcohol: shake 2h in the solvent of water=10: 10: 3, the thalline lipid repeated degreasing 3 times, wrap with aluminium-foil paper, behind the Zha Dong in vacuum drying chamber dry (＜50 ℃).Dried cell is smashed to pieces with clean glass bar; And, carry out ultrasonication (500w, 60S * 90S * 12 time) with minimum broken damping fluid dissolved cell; Solution after the ultrasonication repeats to extract three times with the mixed solution of Triton X-114 and PBS; Merge organic phase, add 10 times of volume 95% ethanol and spend the night collecting precipitation.Add the pronase (available from Shanghai company limited of Roche Group) of 1/10 volume, hatch 24h for 37 ℃, use distill water dialysis 24h again, obtain lipopolysaccharides bullion (sample contains LAM, LM, materials such as PIMs).
15%SDS-PAGE combines silver to dye colour developing evaluation (Shi L, et al.Mol Biol, 2009).
2, the purifying of lipopolysaccharides and evaluation
Choose Sephacryl S-200HiPrep 26/60 series connection Sephacryl S-100HiPrep 16/60 chromatographic column (GE Healthcare company), connect high performance liquid chromatograph (Prominenece UFLC, day island proper Tianjin company).Use preceding with containing 10mM Tris-HCl, the post damping fluid sample dissolution of 0.2M NaCl, upper prop.The adjustment flow velocity is 1ml/min, collects sample with automatic collector, 120/pipe.The method that 15%SDS-PAGE combines silver to dye confirms to contain the lipopolysaccharides pipe, merges LAM and LM sample respectively, obtains LAM and LM behind the purifying.
3, quantitative sugared constituent analysis
(1) anthrone method is measured the content of polysaccharide in the lipopolysaccharides extract
With the anhydrous grape saccharide is the polysaccharide standard, makes the typical curve of concentration range 0～100ug/ml, detects the absorbance of 620nm, the polyoses content in the straight-line regression calculation sample.
(2) discriminating has or not protein contamination:
The a.SDS-PAGE electrophoresis combines the coomassie brilliant blue staining inspection to have or not obvious band to occur
The b.BCA method is measured protein content
4, the lipopolysaccharide molecule amount is identified
Measure LAM, LM molecular weight with ground substance assistant laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOFMS) (autoFLEX, Bruker company), and confirm that molecular weight is respectively about 17KDa and 9.0KDa.
5, use the immunogenicity of ELISA method assessment Much's bacillus CA
(1) with coating buffer antigen LAM and LM are diluted to working concentration, multiple hole is set, encapsulate in 96 hole ELISA Plates, every hole 100 μ l, 4 ℃ are spent the night.
(2) discard coating buffer, cleansing solution washing three times each three minutes, dries on thieving paper.
(3) every hole adds 300 μ l confining liquids, 37 ℃ of incubations 1 hour.
(4) discard confining liquid, cleansing solution washing three times each three minutes, dries on thieving paper.
(5) establish first hole as control wells, every hole adds the serum to be checked of 100 μ l dilution, 37 ℃ of incubations hour.
(6) discard serum, cleansing solution washing three times each three minutes, dries on thieving paper.
(7) every hole adds the goat anti-human igg antibody of the horseradish peroxidase-labeled of 100 μ l dilution, 37 ℃ of incubations 1 hour.
(8) discard liquid, cleansing solution washing three times each three minutes, dries on thieving paper.
(9) every hole adds the substrate solution of the fresh configuration of 100 μ l, 37 ℃ of incubations 10～30 minutes, lucifuge.
(10) every hole adds 50 μ l stop buffer cessation reactions, and 450nm detects the OD value.(assaying reaction carried out with interior after adding stop buffer in 15 minutes.）
6, the making of carbohydrate chip
1) with the epoxy silane/toluene solution infiltration 30min of solid phase carrier, to form the monofilm of epoxide function with 1M;
2) the above-mentioned solid phase carrier that obtains is used α, ω-diamines-polyglycol (M down at 60 ℃
W=2000) (Shanghai Mai Ruier chemical technology company limited) solution-treated makes the polyglycol individual layer molecule that covers one deck band primary amino radical on the solid phase carrier;
3) place the phosphate buffer that contains 1mM tertbutyloxycarbonyl aminooxy group acetate, ethene and N-hydroxy-succinamide to handle 2.5h amidized solid phase carrier, 2h is handled with 2M HCl/ acetic acid in the washing back, makes primary amino radical convert azanol to through chemical modification;
Two kinds of lipopolysaccharides LAM that 4) will from above-mentioned thalline, extract with automatic point sample instrument and LM solution are put successively and are added to surface of solid phase carriers (seeing accompanying drawing 1), lipopolysaccharides are fixed on the solid phase carrier through covalent bond thereby utilize aldehyde radical and the active azanol of lipopolysaccharide molecule reducing end to carry out oximation reaction.
5) got carbohydrate chip in 12 hours in 25-60 ℃ of incubation.
7. the detection of sample
1) directly be added in blood serum sample to be checked on the carbohydrate chip; Antibody is attached on the carbohydrate chip through the specific recognition with lipopolysaccharides, forms specific lipopolysaccharides and antibody complex.
2) hatched 30 minutes for 37 ℃.
3) clean with 1 * lavation buffer solution, remove the non-specific binding material on the carbohydrate chip, disturb to eliminate.
4) add the enzyme labelled antibody working fluid on the carbohydrate chip, 37 ℃ of incubations 30 minutes.
5) clean with 1 * lavation buffer solution, remove the non-specific binding material on the carbohydrate chip, disturb to eliminate.
6) add developer A, B liquid (see table 1 make a bet 2, annotate 3) mixing, put 37 ℃ of lucifuges colour developing 10min.
7) add stop buffer, judged result.
Colour developing result according to reaction judges whether this patient serum sample to be checked is m tuberculosis infection.No matter LAM or LM place have blue spot to occur, and all can be judged as tubercle bacillus affection; Otherwise, locate all not develop the color, be judged to be feminine gender for 2.
8, carbohydrate chip kit composition is seen table 1.
Table 1 carbohydrate chip detection kit is formed
| Detectable | Specification | Quantity |
| Carbohydrate chip (solid phase carrier that combines CA) | Bar | 20 |
| Solution 1 (20 * lavation buffer solution) | The 20ml/ bottle | 1 |
| Solution 2 (serum dilution) | The 20ml/ bottle | 1 |
| Solution 3 (HRP ELIAS secondary antibody) | The 5ml/ bottle | 1 |
| Solution 4 (colour developing liquid A, colour developing liquid B) | The 10ml/ bottle | Each is 1 years old |
| Solution 5 (stop buffer) | The 20ml/ bottle | 1 |
Annotate 1: every box can detect 20 people; Preservation condition: 4 ℃ of preservations.
Solution 1 (20 * lavation buffer solution): 20 * PBST (pH 7.2) 500ml:NaCl 80g, KCl 2g, Na
2HPO
414.4g, KH
2PO
44.8g Tween-20 40ml adds water and is settled to 500ml.
Solution 2 (serum dilution): bovine serum albumin(BSA) (BSA) 0.1g adds 1 * lavation buffer solution and is settled to 100ml.
Solution 3 (HRP ELIAS secondary antibody): the goat anti-human igg antibody of horseradish peroxidase-labeled (R&D company).
Solution 4 (colour developing liquid A, colour developing liquid B): the preparation of 100ml colour developing A liquid: sodium acetate (C
2H
3NaO
2) 2.270g, citric acid (C
6H
8O
7H
2O) 0.320g, 30%H
2O
260ul pH is 5.45; The preparation of 100ml colour developing B liquid: citric acid 0.036g, EDTA-Na
20.172g, glycerine 10.0ml, DMSO 1.5ml, TMB 0.04g
Solution 5 (stop buffer): 2M H
2SO
4
Embodiment 2 utilizes kit to check clinical diagnosis effect lungy, and two kinds of tuberculosis antigen ESAT6 commonly used and CFP10 compare synchronously with in the world.
1, sample:
Mycobacterium clinical patient serum or plasma specimen 90 examples (medical treatment center, Wuhan), divide two groups:
Experimental group: active tuberculosis 57 examples, all patients all dynamic observe and make a definite diagnosis through bacteriology, iconography and clinical symptoms.
Control group: be non-tuberculosis patient and normal healthy people, totally 33 examples comprise pneumonia, chronic bronchial pneumonia, asthma, lung cancer and normal health examinee.Non-tuberculosis patient is all made a definite diagnosis according to the effect of various inspection method inspections and clinical treatment.
2, method:
Detect with the serum of patent kit of the present invention above-mentioned experimental group and control group.Contrast with homemade Much's bacillus specific antigen ESAT6 and CFP10 synchronously.
Remarks:
Self-control recombinant antigen ESAT6 and CFP10 albumen: with Much's bacillus type strain H
37Rv pnca gene group DNA is a template; The Auele Specific Primer according to albumen with according to restriction enzyme site retrieval and design of primers principle and application software Primer 5.0 designs makes up pET21a-esat6 or Pet32a-cfp10 recombinant plasmid by conventional gene engineering method, changes e. coli bl21 (DE3) physS expression strain over to, and 1mM IPTG induces 3h can obtain a large amount of recombinant proteins for 37 ℃; Its purity of nickel post affinity purification reaches more than 90%; Recombinant protein is used 20mM Tris-HCl, pH 8.0 dialysis back ultrafiltration and concentration, the BCA method detects protein concentration; Press the packing of 1mg/ pipe, freeze-drying is preserved.
The carbohydrate chip testing result is judged: no matter LAM or LM place have blue spot to occur, and can be judged as positive findings is the infection of tuberculosis mycomycete; Otherwise, locate all not develop the color, be judged to be feminine gender for 2.
3, result
Table 2 is distinguished based on carbohydrate chip and ESAT-6, the CFP-10 antigen of LAM, LM antigen
To normal healthy controls serum and tuberculosis patient serum IgG reaction result (case load: 90)
Table 2 result shows: compare with CFP10 with two kinds of tuberculosis antigen ESAT6 commonly used in the world, carbohydrate chip detection specificity that makes up based on antigen LAM and LM and single antigen are approaching, but susceptibility has the raising of certain degree.In addition, this research has fast, and is easy, need not specific installation, is fit to characteristics such as clinical practice.In a word, this result of study tentatively shows: carbohydrate chip of the present invention demonstrates good prospects for application in the tuberculosis clinical diagnosis, is worth on probation at clinical expansion, and carries out more deep clinical research.
The above only is a preferred implementation of the present invention; Should be pointed out that for those skilled in the art, under the prerequisite that does not break away from the principle of the invention; Can also make some improvement and retouching, these improvement and retouching also should be regarded as protection scope of the present invention.
Claims (8)
1. carbohydrate chip that is used for tuberculosis rapid diagnosis; It comprises solid phase carrier; It is characterized in that said carbohydrate chip also comprises Much's bacillus liopopolysaccharides antigen probe, said Much's bacillus liopopolysaccharides antigen probe is fixed on the said solid phase carrier through oximation reaction.
2. carbohydrate chip as claimed in claim 1 is characterized in that, said Much's bacillus lipopolysaccharides extracts from Much's bacillus H37Rv bacterial strain.
3. carbohydrate chip as claimed in claim 1 is characterized in that, said Much's bacillus lipopolysaccharides is lipopolysaccharides LAM and LM.
4. carbohydrate chip as claimed in claim 1 is characterized in that, said solid phase carrier is nitrocellulose membrane or the microwell plate through chemical modification.
5. carbohydrate chip as claimed in claim 4 is characterized in that, the chemical modification of said solid phase carrier is to carry out according to following steps:
(1) solid phase carrier is soaked into epoxy silane/toluene solution, form the monofilm of epoxide function;
(2) the above-mentioned solid phase carrier that obtains is used α, ω-diamines-polyglycol (M
W=2000) solution-treated makes the polyglycol individual layer molecule that covers one deck band primary amino radical on the solid phase carrier;
(3) place the phosphate buffer of tertbutyloxycarbonyl aminooxy group acetate, ethene and N-hydroxy-succinamide to handle amidized solid phase carrier, handle with HCl/ acetic acid the washing back, makes primary amino radical convert azanol to through chemical modification.
6. a method for preparing each described carbohydrate chip of claim 1 to 5 is characterized in that, may further comprise the steps:
(1) extraction, purifying Much's bacillus cell membrane liopopolysaccharides antigen;
(2) utilize the method for oximation reaction that the liopopolysaccharides antigen probe is fixed in the solid phase carrier through chemical modification, be made into carbohydrate chip.
7. method as claimed in claim 6 is characterized in that, the chemical modification of said solid phase carrier is to carry out through following steps:
(1) solid phase carrier is soaked into epoxy silane/toluene solution, to form the monofilm of epoxide function;
(2) the above-mentioned solid phase carrier that obtains is used α, ω-diamines-polyglycol (M
W=2000) solution-treated makes the polyglycol individual layer molecule that covers one deck band primary amino radical on the solid phase carrier;
(3) place the phosphate buffer of tertbutyloxycarbonyl aminooxy group acetate, ethene and N-hydroxy-succinamide to handle amidized solid phase carrier, handle with HCl/ acetic acid the washing back, makes primary amino radical convert azanol to through chemical modification.
8. a tuberculosis detection kit is characterized in that, it comprises each described carbohydrate chip of claim 1 to 5.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012101854397A CN102707053A (en) | 2012-06-07 | 2012-06-07 | Carbohydrate chip for quickly diagnosing tuberculosis, preparation method thereof and tuberculosis diagnostic kit adopting carbohydrate chip |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012101854397A CN102707053A (en) | 2012-06-07 | 2012-06-07 | Carbohydrate chip for quickly diagnosing tuberculosis, preparation method thereof and tuberculosis diagnostic kit adopting carbohydrate chip |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102707053A true CN102707053A (en) | 2012-10-03 |
Family
ID=46899990
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012101854397A Pending CN102707053A (en) | 2012-06-07 | 2012-06-07 | Carbohydrate chip for quickly diagnosing tuberculosis, preparation method thereof and tuberculosis diagnostic kit adopting carbohydrate chip |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102707053A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017211314A1 (en) * | 2016-06-08 | 2017-12-14 | Kei International Limited | Method for detecting presence of mycobacterial material in sample using immobilised mannosyl phosphoketide antigen |
| CN109791145A (en) * | 2016-06-08 | 2019-05-21 | 凯仪国际有限公司 | Use method existing for mycobacteria substance in fixed mannosphosphorylation ketone antigen test sample |
| RU220626U1 (en) * | 2023-08-13 | 2023-09-26 | Алексей Георгиевич Наумов | DEVICE FOR DIAGNOSIS OF TUBERCULOSIS INFECTION |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1164188A1 (en) * | 2000-06-16 | 2001-12-19 | Glaxo Group Limited | Auxotrophic Mycobacterium mutants and tuberculosis vaccine |
| CN101974099A (en) * | 2010-11-15 | 2011-02-16 | 上海市肺科医院 | Purification method of tubercle bacilipin Arab mannan and tubercle bacillus diagnostic reagent |
-
2012
- 2012-06-07 CN CN2012101854397A patent/CN102707053A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1164188A1 (en) * | 2000-06-16 | 2001-12-19 | Glaxo Group Limited | Auxotrophic Mycobacterium mutants and tuberculosis vaccine |
| CN101974099A (en) * | 2010-11-15 | 2011-02-16 | 上海市肺科医院 | Purification method of tubercle bacilipin Arab mannan and tubercle bacillus diagnostic reagent |
Non-Patent Citations (3)
| Title |
|---|
| MIAO TONG等: "A multiplexed and miniaturized serological tuberculosis assay identifies antigens and discriminate maximally between TB and non-TB sera", 《 JOURNAL OF IMMUNOLOGICAL METHODS》 * |
| XICHUN ZHOU等: "Oligosaccharide microarrays fabricated on aminooxyacetyl functionalized glass surface for characterization of carbohydrate-protein interaction", 《BIOSENSORS AND BIOELECTRONICS》 * |
| 甘甜甜等: "糖芯片合成中的固定化策略", 《化学进展》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017211314A1 (en) * | 2016-06-08 | 2017-12-14 | Kei International Limited | Method for detecting presence of mycobacterial material in sample using immobilised mannosyl phosphoketide antigen |
| CN109791145A (en) * | 2016-06-08 | 2019-05-21 | 凯仪国际有限公司 | Use method existing for mycobacteria substance in fixed mannosphosphorylation ketone antigen test sample |
| US11243204B2 (en) | 2016-06-08 | 2022-02-08 | Kei International Limited | Method for detecting the presence of mycobacterial material in a sample using at least two antigens |
| RU220626U1 (en) * | 2023-08-13 | 2023-09-26 | Алексей Георгиевич Наумов | DEVICE FOR DIAGNOSIS OF TUBERCULOSIS INFECTION |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP1381864B1 (en) | Detection of candida | |
| CN101738473B (en) | Treponema pallidum antibody diagnostic kit and preparation method thereof | |
| AU2002248996A1 (en) | Detection of candida | |
| CN101014622B (en) | Enriched antibody for detecting mycobacterial infection, methods of use and diagnostic test employing same | |
| CN105137072B (en) | A kind of mycobacterium tuberculosis LAM detection kit, preparation method and using method | |
| Xiang et al. | Development of a rapid, sensitive, dye immunoassay for schistosomiasis diagnosis: a colloidal dye immunofiltration assay | |
| CN101701958A (en) | Fluorescent test paper strip capable of simultaneously testing newcastle disease and bird flu virus and application thereof | |
| EP3243076B1 (en) | Methods for detecting a marker for active tuberculosis | |
| He et al. | Development of a lateral flow immunoassay for the rapid diagnosis of invasive candidiasis | |
| US20230358757A1 (en) | Antigen for 2019 novel coronavirus and detection use thereof | |
| CN101592664A (en) | Hepatitis B e antibody magnetic particle chemiluminescent immunoassay kit and preparation method thereof | |
| CN102707053A (en) | Carbohydrate chip for quickly diagnosing tuberculosis, preparation method thereof and tuberculosis diagnostic kit adopting carbohydrate chip | |
| CN101315379A (en) | Reagent kit for detecting Ractopamine and application thereof | |
| Tel et al. | Development of lateral flow test for serological diagnosis of tularemia | |
| CN108254555A (en) | A kind of fluorescence immunoassay for SFTSV detections detects card and its preparation method and application | |
| CN101377494B (en) | Chemiluminescence immune analytic reagent kit for detecting tuberculosis antibody | |
| CN102887944A (en) | Chlamydia pneumonia antigen, method for preparing antigen, fast detection method and reagent for detecting anti-chlamydia pneumonia antibody by utilizing antigen | |
| CN114720700A (en) | Application of reagent for detecting anti-cytoskeleton-associated protein4-IgG autoantibody in preparation of kit for detecting vascular endothelial injury | |
| JPS61501226A (en) | Methods and devices for concentrating and detecting biomolecules and biological cells | |
| CN101738472A (en) | Biological chip for synchronously detecting typhoid and paratyphoid and preparation method thereof | |
| CN101377499A (en) | Nerve specificity olefinic alcohol enzyme chemiluminescence immune analysis determination reagent kit and preparing method thereof | |
| CN105384805B (en) | A kind of diagnosis polypeptide and its application | |
| CN101592663A (en) | Hepatitis B e antigen magnetic particle chemiluminescent immunoassay kit and preparation method thereof | |
| NL2015553B1 (en) | A method for detecting a marker for active tuberculosis. | |
| CN202066856U (en) | Clenbuterol hydrochloride and ractopamine joint inspection and rapid test card |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20121003 |