CN103667011A - Micro-fluidic chip for loop-mediated isothermal amplification, preparation method and application of micro-fluidic chip - Google Patents
Micro-fluidic chip for loop-mediated isothermal amplification, preparation method and application of micro-fluidic chip Download PDFInfo
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Abstract
The invention discloses a micro-fluidic chip for loop-mediated isothermal amplification, preparation method and application of the micro-fluidic chip. The micro-fluidic chip comprises a substrate layer and a functional layer stacked on the substrate layer, the functional layer is provided with at least one micro-fluidic structure unit, each micro-fluidic structure unit comprises an injection port, a reaction area and a micro-fluidic pipeline for connecting the injection port with the reaction area. A plurality of micro-fluidic structure units are integrated on the micro-fluidic chip, each micro-fluidic structure unit is capable of independently performing loop-mediated isothermal amplification, therefore, the micro-fluidic chip can be used for performing flux operation on the loop-mediated isothermal amplification and performing a plurality of loop-mediated isothermal amplification reactions simultaneously, and the efficiency is greatly improved.
Description
Technical field
The present invention relates to foranalysis of nucleic acids detection technique field, relate in particular to a kind of micro-fluidic chip for ring mediated isothermal amplification, and its preparation method and application.
Background technology
According to statistics, the practical application approximately 60% of biotechnology is aspect medical and health at present, and medical biotechnology is healthy to protect mankind, improving the quality of living produces huge pushing effect.Gene diagnosis (gene diagnosis) is exactly according to the pairing of nucleic acid molecule base complementrity and sex change, renaturation principle, utilize the technological method (as gene probe hybridization, gene amplification in vitro, electrophoresis, DNA sequencing, difference demonstration etc.) of modern molecular biology and molecular genetics, at DNA or rna level, by detecting the abnormal of the existence of Disease-causing gene (native gene or foreign pathogens gene), the defect of gene structure or genetic expression, thus the method that disease is judged.
Diseases induced reason is the cause of disease, and the object of diagnosis is just to locate the cause of disease, wherein the diagnosis of infectious diseases has roughly been experienced diagnosis, immunology diagnosis and gene diagnosis such process of clinical medicine diagnosis, cell levels.Especially the development of modern biotechnology has applied to diagnostics by immunology and molecular biological technology, has solved a difficult problem for traditional diagnosis, makes diagnostics obtain tremendous development.From American scientist in 1978, adopt first DNA restriction fragment length polymorphism (restriction fragment length polymorphism, RFLP) technology, since the prenatal gene diagnosis that fetus is carried out to sickle-cell anemia disease by amniocyte succeeds, the modern molecular biology technique of take is that the gene diagnosis technology of relying on has obtained fast development.At present, gene diagnosis has developed into the diagnostic techniques of effect for lifting clinically.Compare with immunology diagnosis technology, gene diagnosis has salient feature and advantage as state-of-the art diagnostic techniques, nucleic acid detection technique has the features such as pathogenic agent that high specificity, susceptibility are high, do not need operation to live, and in the detection of infectious disease pathogens, application is more and more extensive.Isothermal amplification technology is under a kind of steady temperature, to carry out the technology of augmentation detection, and its amplification efficiency is higher, the time is short and have higher susceptibility and specificity, is the instant technology detecting in a kind of applicable scene.Loop-mediated isothermal amplification technique (loop-mediated isothermal amplification, LAMP) be a kind of isothermal amplification technology, in this technology, need to realize the amplification to same target compound by 4 to 6 primers, thereby greatly increase the specificity of amplification; Susceptibility aspect, can detect the target molecule that is low to moderate 1 copy in this technical know-how.But this technology, due to many primers of needs, cannot realize the Multiple detection in same reaction system, has limited the application in flux context of detection.
Micro-fluidic chip technology is that the basic operation units such as the sample preparation of biology, chemistry and medical analysis process, reaction, separation and detection are integrated on the chip of a micro-meter scale, automatically completes analysis whole process.Due to its great potential in fields such as biology, chemistry and medical science, developed into the brand-new research field of the subject crossing such as a biology, chemistry, medical science, fluid, electronics, material and machinery.On micro-fluidic chip, can carry out a lot of continuous parallel analyses, aspect the microminiaturization of analyzing and flux, there are great potentiality.
Therefore,, by significant in micro-fluidic chip technology introducing ring mediated isothermal amplification, this just needs the corresponding micro-fluidic chip for ring mediated isothermal amplification.
Summary of the invention
The present invention is directed to ring mediated isothermal amplification and be difficult to the defect that flux detects, a kind of micro-fluidic chip for ring mediated isothermal amplification is provided, integrated a plurality of micro-fluidic structures unit on this micro-fluidic chip, each micro-fluidic structure unit can carry out separately ring mediated isothermal amplification, therefore use this micro-fluidic chip to carry out flux operation to ring mediated isothermal amplification, carry out a plurality of loop-mediated isothermal amplifications simultaneously, greatly improve efficiency, promoted development and the application of loop-mediated isothermal amplification technique.
The invention provides technical scheme:
In first aspect, the invention provides a kind of micro-fluidic chip for ring mediated isothermal amplification, comprise stratum basale and the functional layer stacked with described stratum basale, described functional layer arranges at least one micro-fluidic structure unit, and each micro-fluidic structure unit comprises the micro-fluidic pipeline of the described injection port of injection port, reaction zone and connection and reaction zone.
Preferably, the quantity of described injection port is 2, is separately positioned on the both sides of reaction zone, with reaction zone and micro-fluidic pipeline point-blank.
In micro-fluidic chip of the present invention, the shape of described reaction zone can be the different shapes such as cylindrical, rectangular parallelepiped, pref. cylindrical.
In micro-fluidic chip of the present invention, described micro-fluidic pipeline can be the micro-fluidic pipeline of linear pattern or the micro-fluidic pipeline of nonlinear type, preferably the micro-fluidic pipeline of linear pattern.
In micro-fluidic chip of the present invention, described injection port opening is in functional layer on the surface away from a side of stratum basale.
In micro-fluidic chip of the present invention, the material of described stratum basale and/or functional layer can be quartz, glass, polycarbonate (Polycarbonate, PC), polymethylmethacrylate (Polymethyl Methacrylate, PMMA) or polydimethylsiloxane (Polydimethylsiloxane, PDMS).Stratum basale can be identical with the material of functional layer, also can be different.Can also carry out activation treatment to the material of stratum basale, such as plasma bombardment and/or acid treatment etc., make it with covalently cross-linked biomolecules.Surface-closed processing can be carried out with encapsulant in micro-fluidic pipeline and reaction zone, the adsorptivity of the internal surface that reduces pipeline to biomolecules, and then being adsorbed by pipeline of making that material to be checked in sample can less amount, thus improve detection sensitivity.
In micro-fluidic chip of the present invention, together with described stratum basale fits tightly with described functional layer.Particularly, together with described stratum basale can fit tightly by chemical bond or binding agent with described functional layer.Described chemical bond can be by plasma bombardment, siloxane bond to be opened to form hydroxyl and the chemical bond that condensation reaction generates occurs at interface; Described binding agent can be one or more in epoxy resin, urethane, polystyrene, polyacrylic ester, ethylene-vinyl acetate copolymer.
In micro-fluidic chip of the present invention, described functional layer can prepare by the method for mechanical workout, molding or etching.These methods are all the technology of comparative maturity, such as specifically can be with reference to the processing technology > > of < < microfluidic analysis chip, Yin Xuefeng, Fang Qun, reach the clouds and raise, Modern Scientific Instruments, 04 phase of calendar year 2001.
In micro-fluidic chip of the present invention, the long 50-80mm of described stratum basale, for example 55mm, 60mm, 65mm, 70mm, 75mm or 80mm, preferably 75mm; Wide 10-30mm, for example 10mm, 15mm, 20mm, 25mm or 30mm, preferably 25mm; Thick 1-2mm, for example 1mm, 1.2mm, 1.5mm, 1.8mm or 1.9mm, preferably 1mm.
In micro-fluidic chip of the present invention, the wide 0.1-0.7mm of the micro-fluidic pipeline of described linear pattern, for example 0.1mm, 0.2mm, 0.3mm, 0.4mm, 0.5mm, 0.6mm or 0.7mm, preferably 0.5mm; Dark 0.1-0.7mm, for example 0.1mm, 0.2mm, 0.3mm, 0.4mm, 0.5mm, 0.6mm or 0.7mm, preferably 0.5mm.
In micro-fluidic chip of the present invention, described cylindrical reaction zone radius 0.1-1.0mm, for example 0.1mm, 0.2mm, 0.3mm, 0.4mm, 0.5mm, 0.6mm, 0.7mm, 0.8mm, 0.9mm or 1.0mm, preferably 0.5mm; High 0.5-3mm, for example 0.6mm, 0.8mm, 1.0mm, 1.2mm, 1.5mm, 1.8mm, 2.2mm, 2.5mm or 2.8mm, preferably 2mm.
In micro-fluidic chip of the present invention, described injection port bottom radius 0.2-0.3mm, for example 0.2mm, 0.22mm, 0.24mm, 0.26mm or 0.28mm, preferably 0.25mm; Top radius 0.6-1.0mm, for example 0.6mm, 0.7mm, 0.8mm, 0.9mm or 1.0mm, preferably 0.8mm.
In second aspect, the invention provides a kind of preparation method of the micro-fluidic chip as described in first aspect, comprise the following steps:
(1) method by mechanical workout, molding or etching prepares functional layer;
(2) the micro-fluidic pipe surface of described functional layer is stacked on stratum basale, by plasma bombardment, forms covalent linkage and connect or fit tightly together by binding agent, form described micro-fluidic chip.
In the third aspect, the invention provides the method that the micro-fluidic chip of a kind of use as described in first aspect carries out ring mediated isothermal amplification, comprise the following steps:
(a) after ring mediated isothermal amplification reagent mix is even, from the injection port of micro-fluidic chip, inject, by micro-fluidic pipeline, enter in reaction zone;
(b) with binding agent, seal injection port, described micro-fluidic chip is placed in thermostatic equipment and reacts setting-up time, then detect also turbidity or the fluorescence in analytical reaction district;
Preferably, in described thermostatic equipment, temperature is 60-68 ℃;
Preferably, described setting-up time is 20-80min.
In fourth aspect, the invention provides the application of a kind of micro-fluidic chip as described in first aspect in ring mediated isothermal amplification.
It should be noted that: third aspect present invention and fourth aspect may relate to the medical diagnosis on disease of medical field; also may relate to the situation of non-medical diagnosis on disease; such as the aspects such as microorganism in testing environment; the protection domain of the claim that in the present invention, this partial content is corresponding is limited with the situation of non-medical diagnosis on disease, gets rid of the situation of medical diagnosis on disease.
Beneficial effect of the present invention is: the micro-fluidic chip for ring mediated isothermal amplification of the present invention, comprise stratum basale and the functional layer stacked with described stratum basale, described functional layer arranges at least one micro-fluidic structure unit, and each micro-fluidic structure unit comprises the micro-fluidic pipeline of the described injection port of injection port, reaction zone and connection and reaction zone.Due to integrated a plurality of micro-fluidic structures unit on this micro-fluidic chip, each micro-fluidic structure unit can carry out separately ring mediated isothermal amplification, therefore use this micro-fluidic chip to carry out flux operation to ring mediated isothermal amplification, carry out a plurality of loop-mediated isothermal amplifications simultaneously, greatly improve efficiency, promoted development and the application of loop-mediated isothermal amplification technique.
Accompanying drawing explanation
Fig. 1 is the perspective view of the micro-fluidic chip for ring mediated isothermal amplification of the present invention, wherein 1 presentation function layer; 2 represent stratum basale; 11 represent injection port; 12 represent cylindrical reaction zone, are the main region that reacts and detect; 13 represent the micro-fluidic pipeline of linear pattern, are communicated with injection port and cylindrical reaction zone.
Fig. 2 is the vertical view of the micro-fluidic chip for ring mediated isothermal amplification of the present invention, and a plurality of micro-fluidic structures unit is arranged side by side in functional layer.
Fig. 3 is the vertical view of the micro-fluidic structure unit of the micro-fluidic chip for ring mediated isothermal amplification of the present invention, and wherein 11 represent injection port; 12 represent cylindrical reaction zone; 13 represent the micro-fluidic pipeline of linear pattern.
Fig. 4 is the micro-fluidic structure unit longitudinal section of the micro-fluidic chip for ring mediated isothermal amplification of the present invention, wherein 1 presentation function layer; 2 represent stratum basale; 11 represent injection port; 12 represent cylindrical reaction zone; 13 represent the micro-fluidic pipeline of linear pattern.
Fig. 5 is ring mediated isothermal amplification experimental result in the embodiment of the present invention 1, and ordinate zou is turbidity changing value.
Embodiment
Below in conjunction with embodiment, embodiment of the present invention are described in detail.It will be understood to those of skill in the art that following examples are only the preferred embodiments of the present invention, so that understand better the present invention, thereby should not be considered as limiting scope of the present invention.For a person skilled in the art, the present invention can have various modifications and variations, within the spirit and principles in the present invention all, any modification of doing, is equal to and replaces or improvement etc., within all should being included in protection scope of the present invention.Experimental technique in following embodiment, if no special instructions, is ordinary method; Experiment material used, if no special instructions, is and is purchased available from routine biochemistry chemical reagent work.
Please refer to Fig. 1-4, make the micro-fluidic chip for ring mediated isothermal amplification, stratum basale 2 adopts common glass slide, long 75mm, wide 25mm, thick 1mm; Functional layer 1 adopts PDMS material, by photoetching technique, makes template, makes functional layer 1, long 60mm, wide 20mm, thick 3mm; The wide 0.5mm of the micro-fluidic pipeline 13 of linear pattern in functional layer 1, height 0.5mm; Cylindrical reaction zone 12 radius 0.5mm, height 2mm; Injection port 11 bottom radius 0.25mm, top radius 0.8mm.Stratum basale 2 upper surfaces and functional layer 1 pipe surface, by plasma bombardment technology, connect by covalent linkage, fit tightly together, form ring mediated isothermal amplification micro-fluidic chip.
The micro-fluidic chip of preparing below by ring mediated isothermal amplification experimental verification aforesaid method can be used in ring mediated isothermal amplification.
Thermo Pol damping fluid and polysaccharase are purchased from NEB company, dNTPs, template nucleic acid fragment, primer purchased from or synthetic spontaneous work biotechnology (Shanghai) limited-liability company, Manganous chloride tetrahydrate is purchased from Beijing chemical reagents corporation, and trimethyl-glycine is purchased from De Gao bio tech ltd, Jinan.
Embodiment 1: ring mediated isothermal amplification experiment
Find out and detect swine influenza virus (Swine influence virus, SIV) NP protein gene fragment: GGGGCATGTCCCAAGTATGTTAAGCAAAACACTCTGAAGTTGGCAACAGGGATGCG GAATGTACCAGAGAAACAAACTAGAGGCATATTCGGCGCAATAGCAGGTTTCATAG AAAATGGTTGGGAGGGAATGATAGACGGTTGGTACGGTTTCAGGCATCAAAATTCT GAGGGCACAGGACAAGCAGCAGATCTTAAAAGCACTCAAGCAGCCATCG(SEQ ID NO:1, Genbank accession number: NM_213976).
Design and synthesize (life work biotechnology (Shanghai) limited-liability company) following 4 primer sequences:
F3:GGGGCATGTCCCAAGTATG（SEQ?ID?NO：2）；
FIP:CTGCTATTGCGCCGAATATGCCTCTTTTGTTGGCAACAGGGATGCGG（SEQ?ID?NO：3）；
BIP:TGGTTGGGAGGGAATGATAGACGGTTTTGCTGCTTGTCCTGTGCCCTC（SEQ?ID?NO：4）；
B3:CGATGGCTGCTTGAGTGC（SEQ?ID?NO：5）。
Mixing following liquid is one: 10 times of Thermo Pol damping fluid 2.5 μ L of sample, 8mmol/L Adlerika 8 μ L, 0.4mmol/L dNTPs1 μ L, 0.5mmol/L Manganous chloride tetrahydrate 1.25 μ L, 1mol/L trimethyl-glycine 5.2 μ L, 0.2 μ mol/L primers F 30.5 μ L, 0.2 μ mol/L primer B30.5 μ L, 2 μ mol/L primers F IP0.5 μ L, 2 μ mol/L primer BIP0.5 μ L, 0.32U/ μ L Bst polysaccharase 1 μ L, template nucleic acid fragment 0.5 μ L.
Mixing following liquid is two: 10 times of Thermo Pol damping fluid 2.5 μ L of sample, 8mmol/L Adlerika 8 μ L, 0.4mmol/L dNTPs1 μ L, 0.5mmol/L Manganous chloride tetrahydrate 1.25 μ L, 1mol/L trimethyl-glycine 5.2 μ L, 0.2 μ mol/L primers F 30.5 μ L, 0.2 μ mol/L primer B30.5 μ L, 2 μ mol/L primers F IP0.5 μ L, 2 μ mol/L primer BIP0.5 μ L, 0.32U/ μ L Bst polysaccharase 1 μ L, template nucleic acid fragment 1 μ L.
Mixing following liquid is three: 10 times of Thermo Pol damping fluid 2.5 μ L of sample, 8mmol/L Adlerika 8 μ L, 0.4mmol/L dNTPs1 μ L, 0.5mmol/L Manganous chloride tetrahydrate 1.25 μ L, 1mol/L trimethyl-glycine 5.2 μ L, 0.2 μ mol/L primers F 30.5 μ L, 0.2 μ mol/L primer B30.5 μ L, 2 μ mol/L primers F IP0.5 μ L, 2 μ mol/L primer BIP0.5 μ L, 0.32U/ μ L Bst polysaccharase 1 μ L, template nucleic acid fragment 1.5 μ L.
Mixing following liquid is four: 10 times of Thermo Pol damping fluid 2.5 μ L of sample, 8mmol/L Adlerika 8 μ L, 0.4mmol/L dNTPs1 μ L, 0.5mmol/L Manganous chloride tetrahydrate 1.25 μ L, 1mol/L trimethyl-glycine 5.2 μ L, 0.2 μ mol/L primers F 30.5 μ L, 0.2 μ mol/L primer B30.5 μ L, 2 μ mol/L primers F IP0.5 μ L, 2 μ mol/L primer BIP0.5 μ L, 0.32U/ μ L Bst polysaccharase 1 μ L, template nucleic acid fragment 2 μ L.
Mixing following liquid is blank one: 10 times of Thermo Pol damping fluid 2.5 μ L, 8mmol/L Adlerika 8 μ L, 0.4mmol/L dNTPs1 μ L, 0.5mmol/L Manganous chloride tetrahydrate 1.25 μ L, 1mol/L trimethyl-glycine 5.2 μ L, 0.2 μ mol/L primers F 30.5 μ L, 0.2 μ mol/L primer B30.5 μ L, 2 μ mol/L primers F IP0.5 μ L, 2 μ mol/L primer BIP0.5 μ L, 0.32U/ μ L Bst polysaccharase 1 μ L.
Mixing following liquid is blank two: 10 times of Thermo Pol damping fluid 2.5 μ L, 8mmol/L Adlerika 8 μ L, 0.4mmol/L dNTPs1 μ L, 0.5mmol/L Manganous chloride tetrahydrate 1.25 μ L, 1mol/L trimethyl-glycine 5.2 μ L, 0.2 μ mol/L primers F 30.5 μ L, 0.2 μ mol/L primer B30.5 μ L, 2 μ mol/L primers F IP0.5 μ L, 2 μ mol/L primer BIP0.5 μ L, 0.32U/ μ L Bst polysaccharase 1 μ L.
Sample one, sample two, sample three, sample four, blank one and blank two are injected into respectively in the respective cylindrical reaction zone of above-mentioned micro-fluidic chip, at injection port, go out to add epoxy resin, pipe interior liquid is sealed.Micro-fluidic chip is placed in to constant temperature oven, hatches 30 minutes for 65 ℃.
Chip detection: in this test experience, if reacted, cylindrical reaction zone there will be white precipitate.Use digital optic fiber sensor (Japanese Keyemce company, model: FS-V31M) turbidity before and after the experiment of reaction zone is measured, result, as Fig. 5, experimental results show that the amount of template is more, and the precipitation of generation is more, and turbidity changing value is larger.
Applicant's statement, the present invention illustrates detailed features of the present invention and detailed method by above-described embodiment, but the present invention is not limited to above-mentioned detailed features and detailed method, do not mean that the present invention must rely on above-mentioned detailed features and detailed method could be implemented.Person of ordinary skill in the field should understand, any improvement in the present invention is selected the selection of the equivalence replacement of component and the interpolation of ancillary component, concrete mode etc., within all dropping on protection scope of the present invention and open scope to the present invention.
Claims (10)
1. the micro-fluidic chip for ring mediated isothermal amplification, it is characterized in that, comprise stratum basale and the functional layer stacked with described stratum basale, described functional layer arranges at least one micro-fluidic structure unit, and each micro-fluidic structure unit comprises the micro-fluidic pipeline of the described injection port of injection port, reaction zone and connection and reaction zone;
Preferably, the quantity of described injection port is 2, is separately positioned on the both sides of reaction zone, with reaction zone and micro-fluidic pipeline point-blank.
2. micro-fluidic chip according to claim 1, is characterized in that, described reaction zone is cylindrical reaction zone;
Preferably, described micro-fluidic pipeline is the micro-fluidic pipeline of linear pattern;
Preferably, described injection port opening is in functional layer on the surface away from a side of stratum basale.
3. micro-fluidic chip according to claim 1 and 2, is characterized in that, the material of described stratum basale and/or functional layer is quartz, glass, polycarbonate, polymethylmethacrylate or polydimethylsiloxane;
Preferably, the material that the material of described stratum basale is activation treatment;
Preferably, described activation treatment is plasma bombardment and/or acid treatment.
4. according to the micro-fluidic chip described in claim 1-3 any one, it is characterized in that, together with described stratum basale fits tightly with described functional layer;
Preferably, together with described stratum basale fits tightly by chemical bond or binding agent with described functional layer;
Preferably, described chemical bond makes siloxane bond open formation hydroxyl and condensation reaction occurs at interface to generate by plasma bombardment;
Preferably, described binding agent is one or more in epoxy resin, urethane, polystyrene, polyacrylic ester, ethylene-vinyl acetate copolymer.
5. according to the micro-fluidic chip described in claim 1-4 any one, it is characterized in that, described functional layer is that the method by mechanical workout, molding or etching prepares.
6. according to the micro-fluidic chip described in claim 1-5 any one, it is characterized in that the long 50-80mm of described stratum basale, preferred 75mm, wide 10-30mm, preferred 25mm, thick 1-2mm, preferred 1mm.
7. according to the micro-fluidic chip described in claim 1-6 any one, it is characterized in that the wide 0.1-0.7mm of the micro-fluidic pipeline of described linear pattern, preferred 0.5mm, dark 0.1-0.7mm, preferred 0.5mm;
Preferably, described cylindrical reaction zone radius 0.1-1.0mm, preferred 0.5mm, high 0.5-3mm, preferred 2mm;
Preferably, described injection port bottom radius 0.2-0.3mm, preferred 0.25mm, top radius 0.6-1.0mm, preferred 0.8mm.
8. the preparation method of the micro-fluidic chip as described in claim 1-7 any one, is characterized in that, comprises the following steps:
(1) method by mechanical workout, molding or etching prepares functional layer;
(2) the micro-fluidic pipe surface of described functional layer is stacked on stratum basale, by plasma bombardment, forms covalent linkage and connect or fit tightly together by binding agent, form described micro-fluidic chip.
9. the micro-fluidic chip of use as described in claim 1-7 any one carries out a method for ring mediated isothermal amplification, it is characterized in that, comprises the following steps:
(a) after ring mediated isothermal amplification reagent mix is even, from the injection port of micro-fluidic chip, inject, by micro-fluidic pipeline, enter in reaction zone;
(b) with binding agent, seal injection port, described micro-fluidic chip is placed in thermostatic equipment and reacts setting-up time, then detect also turbidity or the fluorescence in analytical reaction district;
Preferably, in described thermostatic equipment, temperature is 60-68 ℃;
Preferably, described setting-up time is 20-80min.
10. the application of the micro-fluidic chip as described in claim 1-7 any one in ring mediated isothermal amplification.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104593255A (en) * | 2015-02-06 | 2015-05-06 | 大连医科大学附属第二医院 | Microfluidic chip for instantly detecting EGFR (epidermal growth factor receptor) mutation |
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| CN105695633B (en) * | 2016-01-29 | 2019-07-16 | 中国人民解放军疾病预防控制所 | Hand-foot-and-mouth disease pathogen detection primer sets and kit |
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| CN107513495A (en) * | 2017-09-01 | 2017-12-26 | 中国科学院苏州生物医学工程技术研究所 | Multichannel droplet detection chip for detection of nucleic acids |
| CN111662804A (en) * | 2020-06-17 | 2020-09-15 | 清华大学 | Digital loop-mediated isothermal amplification micro-fluidic chip for food microorganism detection |
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