CN101487047A - Method for detecting Vibrio cholerae O1 by suspending chip technology - Google Patents
Method for detecting Vibrio cholerae O1 by suspending chip technology Download PDFInfo
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- CN101487047A CN101487047A CNA2008101811963A CN200810181196A CN101487047A CN 101487047 A CN101487047 A CN 101487047A CN A2008101811963 A CNA2008101811963 A CN A2008101811963A CN 200810181196 A CN200810181196 A CN 200810181196A CN 101487047 A CN101487047 A CN 101487047A
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- microballoon
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
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- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention relates to a suspended chip that is used for vibrio cholerae O1 detection and a detection method thereof. The suspended chip comprises a microsphere carrier and an oligonucleotide probe that is fixed on the carrier; wherein, the oligonucleotide probe is a section of DNA sequence of specific gene rfb of the vibrio cholerae O1 that is screened from a vibrio cholerae O-antigen gene cluster. A designed primer is used for augmenting and marking a genomic DNA of a sample to be detected, hybridization is implemented with the suspended chip, and whether vibrio cholerae O1 exists in the sample to be detected can be judged according to hybrid fluorescent intensity. The suspended chip has high detecting sensitivity, can reach the fg level in genomic DNA level and satisfy the detection requirements of clinical samples and environmental samples, is characterized by high specificity, simple operation and low cost, and the like, and has easy popularization. A UNG-Taq enzyme PCR reaction system is adopted so as to greatly reduce PCR false positive results.
Description
Technical field
The present invention relates to a kind of molecular Biological Detection chip, concrete is that a kind of using suspending chip technology detects vibrio cholerae O 1.
Background technology
The suspending chip technology is that biochip technology is combined a kind of new technology that produces with low cytometric analysis, be that a kind of multiple data obtains and analysis platform, full name is " multi-functional many indexs Synchronization Analysis system " (Flexible Multiple Analyte Profiling, xMap), be also referred to as Multi-AnalyteSuspensionArray, organic combination fluorescence-encoded micro-beads, laser technology, micro-fluidic technologies, fast signal handle and data analysis system, promptly guaranteed signal quality, high-throughout molecular detection technology platform of new generation is provided again.The maximum difference of suspending chip and traditional gene chip is, traditional gene chip sample applying is on slide or nylon membrane, rely on coordinate setting to carry out addressing, and suspending chip is covalently bonded to detection probes on the different colour code microballoons, according on the colour code microballoon with fluorescein distinguish.
Suspension chip system mainly is made up of three parts: 1, about microsphere diameter 5.6 μ m, be made of polystyrene, the surface has about 10
8Individual carboxyl site can join with the peptide bond coupling with oligonucleotide probe and albumen; 2, in the fluorescein microballoon mark redness and fluorescent orange element, every kind is divided into 10 concentration gradients, and microballoon is divided into 100 kinds of microballoons that can be identified, and after microballoon is by 635nm laser excitation, can launch the fluorescence of 658nm and 712nm; 3, detector is because of microsphere diameter 5.6 μ m, and by fluorescent mark, adopts the low cytometric analysis principle to detect.
Principle: each colour code microballoon can pass through carboxyl, with specific amalyzing substances (oligonucleotide probe, antigen, antibody etc.) covalent attachment.The microballoon of mark bioprobe is carried out specific the combination with determinand, with the reporter molecules reaction that has the third fluorescein, in a reacting hole, can detect 100 kinds of various objectives molecules in the sample simultaneously again.After the reaction, adopt 96 orifice plates to carry out sample introduction.Detector picks up reaction solution the adding peristaltic pump automatically, pumps in the microcapillary, and under the effect of sheath fluid, the single sense channel that passes through of the microballoon of single distribution.Be provided with two laser probes in the sense channel, a probe can be launched the laser of 635nm wavelength, but two kinds of fluoresceins of excitation labeling microballoon, thus discern different colour code microballoons, carry out qualitative detection; Another laser probe can excite the fluorescein of reporter molecules, is with the reporter molecules fluorescence signal intensity by measuring microballoon, can carry out detection by quantitative.Therefore, suspending chip can carry out the qualitative and quantitative detection target molecule.In sample, contain molecules of interest, molecules of interest and specific microballoon with probe when being attached together, twice laser institute excited fluorescent all can be detected; If do not contain molecules of interest in the sample, then only can detect microballoon with fluorescence.By digital signal processor and the automatic statistical software of computer, analyze the wavelength and the strength of signal of twice laser institute fluorescence excitation, thereby judge several extremely tens of kinds of detection target compounds in the sample to be checked, and, carry out quantitative analysis detecting thing by detecting fluorescence signal intensity.Autopipette adds different samples in the peristaltic pump successively, separates with a bit of gas column between the sample, not only can distinguish different samples like this in continuous analysis process, can be also to prevent the pollution between sample.
Compare suspending chip tool distinct advantages with other protein and nucleic acid detection method:
1) carboxyl of applied range microsphere surface can join with nucleic acid probe and protein molecular coupling, can carry out qualitative, quantitative research to purpose nucleic acid and protein molecular;
2) the susceptibility height as reaction carriers, has increased the reactant contact area with microballoon; Each microsphere surface has about 10
8Individual carboxyl site, with covalent coupling connection oligonucleotide probe, antibody or antigen, the probe density height, the signal of generation is strong; Use fluoroscopic examination, amplified reaction signal, susceptibility improves greatly.
3) biological respinse that good reproducibility carried out carries out in liquid phase environment, be beneficial to biomolecules native conformations such as keeping nucleic acid, albumen, overcome the influence of traditional die steric effect, also be more conducive to the reaction of probe and analyte, improved the accuracy that detects;
4) signal to noise ratio is hanged down in microcapillary single microballoon is detected, and the problem that does not exist the background influence to detect need not wash-out and removes background;
5) high-throughput can carry out qualitative and quantitative analysis to the multiple various objectives molecule in the same sample simultaneously, has promptly improved detection information flux, has saved the sample consumption again;
6) speed of response and detection speed have rapidly and efficiently been improved greatly.Because the biological respinse that is carried out carries out in liquid phase environment, hybridization only needed to finish in 15 minutes, had improved speed of response; In 35-60 minutes, can reach the detection of 100 kinds of indexs respectively simultaneously to 96 different samples; Utilize 96 orifice plates and automatic sample handling system, can carry out the detection of n * 96 sample continuously.
7) the low preparation process of cost need not specific installation, and the reactive polypeptide that is dehydrated into that only carries out between carboxyl and amino gets final product, and is simple; Owing to be the fluid storage mode, once preparation can repeatedly be used, and on average each index detects cost and only needs 2.5-5.0 yuan;
8) be easy to stdn based on These characteristics, make this technology can accomplish stdn, be easy to the development and the popularization of test kit.At present, suspending chip is the biochip that the unique approval of U.S. FDA is used for clinical diagnosis (autoimmune disease detects and the human leucocyte antigen typing).
Cholera is a kind of deadly infectious disease, once causes in history to be very popular for 7 times, has caused massive losses for people's lives and properties.At present, the vibrio cholerae of having identified reaches more than 190 serotype, causes that wherein cholera is pandemic for serotype O1, O139.The vibrio cholerae conventional sense is based on traditional cultural method, complex operation not only, length consuming time, and false negative height.Therefore set up a kind of quick, special, sensitive detecting method is significant to the cholera preventing and controlling.
The analysis of vibrio cholerae O-antigen gene coma information science is found one section region of variability of tool on O-antigen synthase gene rfb can be at this zone screening different serotypes specific sequence (Franciosa G., 1994).
Summary of the invention
Purpose of the present invention aims to provide a kind of suspending chip that detects vibrio cholerae O 1.The suspending chip of detection vibrio cholerae O 1 provided by the present invention comprises the diagnose microballoons and the specificity oligonucleotide probe of microballoon coupling connection therewith, and wherein the oligonucleotide probe sequence is one section sequence of vibrio cholerae O 1 specific gene rfb.The invention also discloses one section oligonucleotide probe sequence, this sequence is 5 '-NH
2-(CH
2)
12-TGGTACAATCACTTCATCGCC-3 ', shown in sequence in the sequence table 3, and 5 ' carry out NH
2-(CH
2)
12Modify.Suspending chip of the present invention in addition also comprises a pair of Auele Specific Primer, and upstream primer is: B-O1rfb-F:5 '-CACGCCACAACAGTATCTAAT-3 ' (see sequence 1 in the sequence table, carry out the Biotin mark); Downstream primer is: O1 rfb-R:5 '-AACGGGTAACGCACCACAC-3 ' (seeing sequence 2 in the sequence table).Above-mentioned primer can increase and comprise one section rfb sequence of above-mentioned probe sequence.
Another object of the present invention is to provide the preparation method of above-mentioned suspending chip, this method may further comprise the steps:
1) preparation oligonucleotide probe;
2) with the amido modified probe of pure water dilution;
3) with the microballoon vibration of deposit, transfer to then in the brown EP pipe;
4) centrifugal collection microballoon adds MES liquid and vibrates;
5) in the suspension microballoon, add above-mentioned probe, and vibration;
6) in microspheres solution, add the new 10mg/ml EDC for preparing, vibration; Carry out incubation then;
7) add the new 10mg/ml EDC that disposes once more in microspheres solution, incubation is carried out in vibration then;
8) in microspheres solution, add 0.02% Tween-20, centrifugal collection microballoon;
9) with the resuspended microballoon of 0.1%SDS, centrifugal collection microballoon; With the TE microballoon that suspends;
10) with the number of blood counting chamber calculating microballoon, 2-8 ℃, keep in Dark Place.
Using suspending chip detection vibrio cholerae O 1 of the present invention carries out by the following method.
According to vibrio cholerae O 1 rfb gene design and the synthetic Auele Specific Primer that is used for pcr amplification, shown in sequence in the sequence table 1 and 2, and primer is wherein carried out Biotin modify; Prepare sample genomic dna to be checked according to a conventional method as template, use above-mentioned Auele Specific Primer to carry out pcr amplification, make Biotin modification on the PCR product band simultaneously; With the suspending chip hybridization of PCR product and preparation, just hybridize with the probe that coupling is associated on the microballoon, the hybridization suspending chip is carried out streptavidin phycoerythrin mark, detect fluorescent signal by flow cytometer then.
Another object of the present invention provides a kind of PCR of overcoming false-positive method.UNG-Taq enzyme system that the present invention adopts has effectively been controlled the PCR false negative and has been produced.Concrete grammar is in the PCR reaction system, and the dTTP amount is reduced by half, and is substituted by dUTP, and adds the UNG (ura DNA glycosidase) of 0.05U.Be impregnated in a certain amount of dUTP in the PCR product, UNG can not be identified as template by interrupting the glycosidic link on the dUTP on the nucleic acid chains, make the nucleic acid that contains dUTP.37 ℃ of incubation 5min of elder generation before operation PCR program make UNG fully digest the PCR product, and during 94 ℃ of template sex change, the UNG inactivation carries out normal PCR reaction.
Suspending chip of the present invention can identify and detect vibrio cholerae O 1, have high specific, highly sensitive, fast, low-cost and easy-to is in characteristics such as popularizations.
Description of drawings
Fig. 1 specificity test-results;
Fig. 2 sensitivity test result;
Fig. 3 UNG-Taq enzyme system digestion PCR product test-results.
Embodiment:
For further specifying the application of the present invention in vibrio cholerae O 1 detects, describe especially exemplified by following preferred embodiment, but application of the present invention is not limited to embodiment.
Embodiment one: primer design and synthetic
1) sequence obtains: by to bunch sequential analysis of vibrio cholerae O-antigen gene, choose O1 specific gene rfb as target sequence, and obtain this gene order from the GenBank public database, be numbered X59554;
2) design primer: adopt Primer Premier 5.0 software design primers, correlation parameter is: 55.0 ℃-59.0 ℃ of Tm values, and GC value 40.0%-60.0%, PCR product size is 100bp-500bp, primer size 22 ± 3bp;
3) primer is selected: the primer of software output is carried out suitable manual regulation, increase or reduce several bases, carry out online Blast comparison at GenBank then, choose the high primer of specificity, and will be wherein one carries out 5 ' terminal Biotin mark.The upstream primer sequence is B-O1rfb-F:5 '-Biotin-CACGCCACAACAGTATCTAAT-3 ', and downstream primer is O1rfb-R:5 '-AACGGGTAACGCACCACAC-3 ', and the amplified fragments size is 153bp;
4) primer is synthetic: the synthetic downstream primer O1rfb-R of worker, the upstream primer B-O1rfb-F of the synthetic Biotin mark of Dalian TakaRa are given birth in Shanghai.
Embodiment two: the design of probe and synthetic
1) sequence obtains: at the non-guiding region designing probe of above-mentioned amplified fragments;
2) designing probe: adopt Primer Premier 5.0 software design probes, selected HybridizationProbes order, designing probe on the Anti-sense chain, parameter is with embodiment one;
3) probe is selected: the probe of software output is carried out suitable manual regulation, increase or reduce several bases, carry out online Blast comparison at GenBank then, choose the high probe of specificity, carry out NH at 5 ' end
2-(CH
2)
12Modify, selected probe sequence is O1rfb-Probe:5 '-NH
2-(CH
2)
12-TGGTACAATCACTTCATCGCC-3 ';
4) probe is synthetic: Dalian TakaRa synthesizes above-mentioned probe.
The preparation method of embodiment three suspending chips
1. dilute amido modified probe to 1mM (lnanomole/ μ l) with pure water;
2. with the microballoon vibration 20sec that lays in;
3. getting 200 μ l microballoons transfers in the brown EP pipe;
4. collection microballoon, 8000g, centrifugal 1-2min;
5. abandon supernatant, add 50 μ l 0.1M MES (2-[N-Morpholino] ethanesulfonic acid, 2-(N-morpholino) ethyl sulfonic acid) liquid pH4.5, vibration 20sec;
6. the probe that in the suspension microballoon, adds 2 μ l 1mM, vibration 20sec;
7. prepare fresh 10mg/ml EDC (1-ethyl-3-[3dimethylaminopropyl] carbodiimide hydrochloride, 1-ethyl-3-3-dimethylaminopropyl carbodiimide), use dH
2O;
8. the 10mg/ml EDC that in microspheres solution, adds the new configuration of 2.5 μ l, vibration;
9. incubation 30min under the room temperature dark condition;
10. prepare fresh 10mg/ml EDC once more, use dH
2O;
11. in microspheres solution, add the 10mg/ml EDC of the new configuration of 2.5 μ l, vibration;
12. incubation 30min under the room temperature dark condition;
13. in microspheres solution, add 1.0mL 0.02% Tween-20;
14.8000 the centrifugal 1-2min of * g collects microballoon;
15. abandon supernatant, with the resuspended microballoon of 1.0mL 0.1% SDS;
16.8000 the centrifugal 1-2min of * g collects microballoon;
17. with 100 μ l TE, pH=8.0, the suspension microballoon,
18. calculate the number of microballoon with blood counting chamber
19. with 2-8 ℃ of microballoons, it is standby to keep in Dark Place.
Embodiment four: suspending chip rapid detection vibrio cholerae O 1 method
1) TIANamp Bacteria DNA kit test kit extracts sample genomic dna to be checked.
2) pcr amplification purpose fragment, reaction conditions is as follows:
The PCR response procedures is: 94 ℃ of sex change 3min; Move 35 circulations: 94 ℃ of 30sec, 53 ℃ of 30sec, 72 ℃ of 40sec; At last, 72 ℃ are extended 3min.
3) hybridization: hybridization solution is formed: 33.3 μ l, 1.5 * TMAC (tetramethylammonium chloride, tetramethylammonium chloride), and the above-mentioned PCR product of 5 μ l adds 5000 microballoons that join with the probe coupling, and 1 * TE adds to 50 μ l with volume.Hybridization solution at 95 ℃ of sex change 4min, is hybridized more than the 15min for 52 ℃.
4) detect: the solution after the above-mentioned hybridization is all transferred in the 96 hole filter plates, microballoon is collected in vacuum filtration, with the resuspended microballoon of 2 μ g/ml S-R-PE (Streptavidin-R-phycoerythrin, streptavidin phycoerythrin), put 52 ℃ of 10min, Luminex detects.
Embodiment five: the specificity checking of suspending chip
The specificity test:
Adopt embodiment four directions method, 45 strain bacterial strains are carried out the specificity checking, the result shows the vibrio cholerae O 1 result that is positive, and other Non-cholera vibrio O1 is all negative, and specificity reaches 100%.
The bacterial strain specifying information is as follows:
Sensitivity test:
Carry out sensitivity test by the method among the embodiment three, congratulate the IEM/PLA860084 research object with dysentery will, the template of extracting is carried out quantitatively, carry out 10 times of dilutions successively, the template amount that adds in the sensitivity test is respectively 2.31fg, 23.1fg, 231fg, 2.31pg, 23.1pg, 231pg, does negative control with pure water.The result shows that sensitivity reaches 2.31fg (being equivalent to 2-3 bacterial counts), can satisfy clinical sample and environmental samples and detect needs.
Embodiment six: it is test that the UNG-Taq enzyme system is eliminated the PCR product pollution
Carry out this test by the method among the embodiment three.The PCR product is carried out quantitatively getting 10 respectively
7, 10
8, 10
9, 10
10, 10
11, 10
12Individual PCR product copy number adopts UNG-Taq enzyme system to react as template, with the negative contrast of pure water, with the positive contrast of the full genome of carried, establishes no UNG, 0.05U UNG respectively as a comparison.The result shows that UNG-Taq enzyme system can digest 10
8Individual PCR product copy number has effectively overcome the PCR false positive and has produced.
Sequence table
＜110〉Inst. of Epidemiology and Microbiology, Academy of Military Medical Sciences, PL
＜120〉a kind of method for detecting Vibrio cholerae O 1 by suspending chip technology
<140>
<141>
<160>3
<210>1
<211>21
<212>DNA
＜213〉artificial sequence
<400>1
<210>2
<211>19
<212>DNA
＜213〉artificial sequence
<400>2
<210>3
<211>21
<212>DNA
＜213〉artificial sequence
<400>3
Claims (8)
1. one kind is used for the suspending chip that vibrio cholerae O 1 detects, comprise diagnose microballoons and with the oligonucleotide probe of diagnose microballoons coupling connection, its feature is one section sequence on the vibrio cholerae O 1 O-antigen synthase gene rfb at this oligonucleotide probe.
2. the suspending chip that detects according to the described vibrio cholerae O 1 of claim 1, wherein oligonucleotide probe be sequence shown in sequence in the sequence table 3, its 5 ' carry out NH
2-(CH
2)
12Modify.
3. the suspending chip that detects according to claim 1 or 2 described vibrio cholerae O 1s is characterized in that this chip can identify and detect vibrio cholerae O 1.
4. the suspending chip that detects according to claim 1 or 2 described vibrio cholerae O 1s is characterized in that comprising a pair of Auele Specific Primer, and primer sequence and carries out 5 ' Biotin to sequence 1 and modifies shown in sequence in the sequence table 1 and 2.
5. claim 1 or the 2 described preparation methods that are used for the suspending chip of Shigellae detection comprise the steps: to prepare oligonucleotide probe; With the amido modified probe of pure water dilution; With the microballoon vibration of deposit, transfer to then in the brown EP pipe; Centrifugal collection microballoon adds MES liquid and vibrates; In the suspension microballoon, add above-mentioned probe, and vibration; The EDC solution that in microspheres solution, adds new preparation, vibration; Carry out incubation then; Add the 10mg/ml EDC of new configuration once more in microspheres solution, incubation is carried out in vibration then; In microspheres solution, add 0.02%Tween-20, centrifugal collection microballoon; With the resuspended microballoon of 0.1%SDS, centrifugal collection microballoon; With the TE microballoon that suspends; Number with blood counting chamber calculating microballoon, keeps in Dark Place by 2-8 ℃.
6. claim 1 or 2 describedly is used for the suspending chip using method that vibrio cholerae O 1 detects, and may further comprise the steps:
A), and primer is wherein carried out Biotin modify according to vibrio cholerae O 1 rfb gene design and the synthetic Auele Specific Primer that is used for pcr amplification;
B) sample genomic dna to be checked of preparation according to a conventional method, and use primer in the step a) to treat this genomic dna of sample and carry out pcr amplification makes simultaneously that Biotin modifies on the PCR product band;
C) suspending chip described in PCR product in the step b) and the claim 1 is hybridized;
D) hybridization suspending chip in the step c) is carried out streptavidin phycoerythrin mark, and it is detected fluorescent signal.
7. a kind of method that detects vibrio cholerae O 1 according to claim 4 is characterized in that above-mentioned steps b) the PCR reaction system set up is as follows:
Form 1 PCR reaction system
The PCR response procedures is: 37 ℃ of incubation 5min; 94 ℃ of sex change 3min; Move 35 circulations: 94 ℃ of 30sec, 53 ℃ of 30sec, 72 ℃ of 40sec; At last, 72 ℃ are extended 3min.
8. a kind of method that detects vibrio cholerae O 1 according to claim 6, wherein hybridization temperature is 52 ℃, hybridization time is greater than 15min.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2528099C2 (en) * | 2013-06-25 | 2014-09-10 | Федеральное казенное учреждение здравоохранения "Российский научно-исследовательский противочумный институт "Микроб" Федеральной службы по надзору в сфере защиты прав потребителей и благополучия человека ("РосНИПЧИ "Микроб") | Biological microchip for detection and multiparameter analysis of anticholeraic antibodies |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1683565A (en) * | 2005-03-15 | 2005-10-19 | 中国人民解放军军事医学科学院放射医学研究所 | Oligonucleotide probe kit for detecting common intestine trac kpathogenic bacteria and its use |
| CN1766615A (en) * | 2005-10-11 | 2006-05-03 | 山东省医药生物技术研究中心 | Liquichip for parallel detection of colorectal cancer protein marker, preparation and application thereof |
| CN101045944A (en) * | 2007-01-12 | 2007-10-03 | 北京爱普益生物科技有限公司 | Gene chip for detecting six kinds of diarrhea pathogens and its prepn process and kit |
| CN101113476A (en) * | 2007-05-30 | 2008-01-30 | 中国疾病预防控制中心传染病预防控制所 | Pathogenic microorganism DNA detecting chip and preparation method and application thereof |
| CN101178403A (en) * | 2007-11-26 | 2008-05-14 | 广州益善生物技术有限公司 | Liquid phase chip used for detecting liver fibrosis and method of producing the same |
-
2008
- 2008-11-27 CN CN2008101811963A patent/CN101487047B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1683565A (en) * | 2005-03-15 | 2005-10-19 | 中国人民解放军军事医学科学院放射医学研究所 | Oligonucleotide probe kit for detecting common intestine trac kpathogenic bacteria and its use |
| CN1766615A (en) * | 2005-10-11 | 2006-05-03 | 山东省医药生物技术研究中心 | Liquichip for parallel detection of colorectal cancer protein marker, preparation and application thereof |
| CN101045944A (en) * | 2007-01-12 | 2007-10-03 | 北京爱普益生物科技有限公司 | Gene chip for detecting six kinds of diarrhea pathogens and its prepn process and kit |
| CN101113476A (en) * | 2007-05-30 | 2008-01-30 | 中国疾病预防控制中心传染病预防控制所 | Pathogenic microorganism DNA detecting chip and preparation method and application thereof |
| CN101178403A (en) * | 2007-11-26 | 2008-05-14 | 广州益善生物技术有限公司 | Liquid phase chip used for detecting liver fibrosis and method of producing the same |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2528099C2 (en) * | 2013-06-25 | 2014-09-10 | Федеральное казенное учреждение здравоохранения "Российский научно-исследовательский противочумный институт "Микроб" Федеральной службы по надзору в сфере защиты прав потребителей и благополучия человека ("РосНИПЧИ "Микроб") | Biological microchip for detection and multiparameter analysis of anticholeraic antibodies |
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