CN106636383A - Detection method of nucleic acid in micro sample - Google Patents
Detection method of nucleic acid in micro sample Download PDFInfo
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- CN106636383A CN106636383A CN201611137387.0A CN201611137387A CN106636383A CN 106636383 A CN106636383 A CN 106636383A CN 201611137387 A CN201611137387 A CN 201611137387A CN 106636383 A CN106636383 A CN 106636383A
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- pcr
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6804—Nucleic acid analysis using immunogens
Abstract
The invention relates to a detection method of nucleic acid in a micro sample. The detection method includes acquiring a composite capture body comprising a magnetic core and a shell covering the magnetic core, wherein the shell is coupled with an antibody which has specific affinity to specific epitope of cells or viruses in the sample; putting the sample in a container, and contacting the sample with the composite capture body to enable the composite capture body to combined with the cells or viruses; applying a magnetic field to the container to adsorb the composite capture body to the inner wall of the container, and removing waste liquid; adding rinsing liquid to wash the composite capture body to obtain a composite capture body-cell combination or composite capture body-virus combination, and removing waste liquid; amplifying and detecting the composite capture body-cell combination or composite capture body-virus combination. By the detection method, the problems of complexity in operation of biological element separation and detection and low specificity in the prior art are solved, and automation and high-throughput treatment of separation and detection processes are realized easily.
Description
Technical field
The present invention relates to field of nucleic acid detection, and in particular to the detection method of micro-example amplifying nucleic acid.
Background technology
From round pcr last century the eighties appearance since, its molecular biology association area application increasingly
Extensively.Especially in recent years, with the raising that people are required clinical diagnosises precision sensitivity, and Personalized medicine concept
Propose and promote, round pcr by its it is quick in clinical diagnosises, sensitive, special the advantages of, be widely applied to rapidly
Clinic diagnosis, by detecting clinical samples in gene, so as to helping doctor to determine medication and therapeutic scheme.
And sample nucleic acid extraction is the premise that round pcr is used, human peripheral blood is in people's vivo immunization reaction and metabolism
Play an important role, can be used for the molecular monitoring of blood system and other numerous systemic diseases, because it is clinically sampled
It is simple and easy to get, it is most widely used in clinical diagnosises or research.
At present whole blood method for extracting nucleic acid mainly has phenol chloroform method, centrifugal column method, paramagnetic particle method.Wherein phenol chloroform method
The complex operation and organic solvent containing big toxicity is rare uses；Centrifugal column method remains current most popular whole blood core
Sour extracting method, but it needs special pellosil chromatographic column, and need ability eluting after eccentric cleaning repeatedly to obtain nucleic acid；
Paramagnetic particle method obtains nucleic acid using magnetic bead absorption, cleaning eluting；Need repeatedly to be centrifuged in these method operating process, change pipe, bar magnet
Separation process, complex operation, extraction time is longer, higher to operation requirement, is unfavorable for clinic diagnosis or research application.
Patent CN201410010668.4 discloses a kind of method for carrying out whole blood genome rapid amplifying, the whole blood base
Because the method for group rapid amplifying is realized by nucleic acid releasing agent and matched PCR reactant liquors, mainly comprise the following steps,
Take 5 μ l nucleic acid releasing agents to be added in PCR reaction tubes, then take 5 μ l blood and be mixed with, room temperature static 5-10 minutes, then
Take the PCR reactant liquors that 40 μ l configure to be directly added in above-mentioned mixed liquor, finally go up machine amplification.The invention is in nucleic acid releasing agent
Under effect, nucleic acid can be direct, quickly discharges from the nucleated cell such as blood middle leukocytes or tiny cell, meanwhile,
Nucleic acid releasing agents can result in haemachrome and other impurities albuminous degeneration again, reduce these interfering materials for follow-up PCR expands
The interference of increasing, so as to realize that poba gene group rapid PCR amplification is detected.
But the method directly discharges the nucleic acid in whole blood from nucleus, virus, the antibacterial that can be introduced in blood
Nucleic acid, and can not fully erased haemachrome and other protein, the presence of these materials can produce dry to subsequent PCR amplification
Disturb, affect the specificity of PCR amplifications.
The content of the invention
It is an object of the invention to solve biotic component in prior art separate and detection process complex operation, specifically
The not high problem of property.
It is a further object of the invention that realizing separating and detection process automatization, high flux process.
The purpose of the present invention is achieved through the following technical solutions：
According to one side according to the present invention, separate and biotic component that may be present in cell in detection sample or virus
Method, including：A. compound capture body is obtained, the compound capture body is comprising magnetic core and is coated in outside the magnetic core
Shell, the shell and antibody coupling, the antibody has special to the cells in sample or the defined epitope in virus
Property affinity；B. the sample is placed in a container and contacts the sample with the compound capture body；C. to described
Container applies magnetic field, the capture complex is adsorbed onto on the container inner wall, removes waste liquid；D. rinsing liquid is added to described
Capture complex washed, if the biotic component exist, obtain capture complex-cell conjugates or capture complex-
Viral conjugates, remove waste liquid；E. amplification process is performed, it is compound to the capture complex-cell conjugates or capture
Nucleic acid in body-viral conjugates is expanded or detected, it is described capture complex to the nucleic acid amplification or detection process not
Produce inhibitory action；. carry out testing result judgement.
Further, the method for the nucleic acid amplification selected from polymerase chain reaction (PCR), reverse transcription PCR, nest-type PRC,
Multiplex PCR, semiquantitive PCR, real-time fluorescence quantitative PCR, In situPCR, ligase chain reaction, randomly amplified polymorphic DNA, isothermal
One or more in amplification technique, recombinant PCR, anchor PCR, asymmetric PCR, inverse PCR or immuno-PCR.
Further, the nucleic acid isothermal amplification technology is untwisted selected from chain replacement amplification technique, rolling circle amplification, dependence
Enzymatic nucleic acid amplification, dependence amplification of nucleic acid sequences, ring mediated isothermal amplification, single primer isothermal amplification technique, rapid isothermal detection are put
One or more in big technology and Q β reproduction technologies..
According to another aspect according to the present invention, the sample is placed in front that the sample is multiple with described in the container
The sample is contacted with the compound capture body after closing the contact of capture body or the sample being placed in the container.
According to another aspect according to the present invention, it is additionally included in before the sample is contacted with the compound capture body to institute
State the step of adding buffer in sample.
According to another aspect according to the present invention, the sample selected from lymph fluid, tear, perspiration, urine, saliva, whole blood,
In cerebrospinal fluid, lung cavity liquid, peritoneal fluid, the liquid in joint, amniotic fluid, interstitial fluid, tissue culture medium or cell culture fluid one
Plant or various.
According to another aspect according to the present invention, the cell selected from human body cell, zooblast, plant cell, antibacterial,
One or more in fungal cell or reconstitution cell.
Preferably, the human body cell is selected from Colonic exfoliative cells, exuviation cell, phagocyte, leukocyte, dry thin
One or more in born of the same parents, neurocyte.
According to another aspect according to the present invention, the antibody is selected from CD45 specific antibodies, E.coli antibody, mAb
One or more in RE-4D8, anti-M13 antibody, influenza A viruss nucleoprotein monoclonal antibody.
According to another aspect according to the present invention, nucleic acid detection method includes：A. using in the separation and detection sample
The method of biotic component that may be present obtains the cell or virus combined on the capture complex, institute in cell or virus
State cell or virus is contained within nucleic acid；B. other amplifications such as PCR amplifications and isothermal duplication or detection mode are performed, obtains described
Cell or viral nucleic acid amplified production or testing result, the capture complex is to PCR amplifications, isothermal duplication or other detections
Mode does not produce inhibitory action；C. the nucleic acid amplification product, isothermal duplication product etc. is checked or analyzed.
According to another aspect according to the present invention, the method that the testing result judges selected from gel electrophoresiss, nucleic acid sequencing,
One or more in making nucleic acid molecular hybridization, the capture complex is to expanding and detecting unrestraint effect.
According to another aspect according to the present invention, the device of cells in sample or virus is separated, including：A. it is combined capture
Body, the compound capture body comprising magnetic core and be coated in the magnetic core outward and antibody coupling shell, the antibody
There is specific affinity to the cells in sample or the defined epitope in virus；B. the sample is made with the compound capture
The device of body contact；C. magnetic field generation device.
According to another aspect according to the present invention, the device with cell in detection sample or viral nucleic acid is separated, including：
A. be combined capture body, the compound capture body comprising magnetic core and be coated in the magnetic core it is outer and antibody coupling outside
Shell, the antibody has specific affinity to the cells in sample or the defined epitope in virus；B. make the sample with
The device of the compound capture body contact；C. magnetic field generation device；D. the cells in sample or the nucleic acid in virus are expanded
Device；E. the device and reagent of the cells in sample or the nucleic acid in virus are detected.
According to another aspect according to the present invention, the device of cell or viral nucleic acid exists in the separation and detection sample
Application in detection of nucleic acids.
Relative to prior art, the beneficial effects of the present invention is：
The compound capture body can specifically bind target cell or virus, and eliminate other in sample it is various into
Point, the biotic component for finally being obtained is exclusive source in target cell or the biotic component of virus；Additionally, in detection of nucleic acids
During, can be prevented effectively from other impurities in sample PCR is expanded and detection of nucleic acids interference.
Further, the method and apparatus according to the present invention used in the sample of different scales, particularly on a small quantity
And micro-example, and method and apparatus according to the present invention is easy to automatization and miniaturization, is capable of achieving high flux gene test.
In addition, it is only necessary to carry out detection of nucleic acids by the sample for gathering micro (1-5 microlitre), such as people's whole blood sample is gathered
Can be adult's finger tip blood, baby's Heel blood etc., while being particularly suitable for the detection of rare sample, reduce detection of nucleic acids pair
The requirement of sample collection, reduces the difficulty of sampling.
Specific embodiment
To make purpose, technical scheme and the advantage of the embodiment of the present invention clearer, below will be in the embodiment of the present invention
Technical scheme be clearly and completely described, it is clear that described embodiment is a part of embodiment of the invention, rather than
Whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art are not making creative work premise
Lower obtained every other embodiment, belongs to the scope of protection of the invention.
On the one hand, the present invention relates to separate the side with biotic component that may be present in cell in detection sample or virus
Method, including：A. obtain compound capture body, the compound capture body comprising magnetic core and be coated in outside the magnetic core with it is anti-
The shell that body is coupled, the antibody has specific affinity to the cells in sample or the defined epitope in virus；B. will
The sample is placed in a container and contacts the sample with the compound capture body；C. magnetic field is applied to the container,
The capture complex is adsorbed onto on the container inner wall, remove waste liquid；D. rinsing liquid is added to enter the capture complex
Row washing, if the biotic component is present, obtains capturing complex-cell conjugates or capture complex-viral conjugates, goes
Except waste liquid；E. detection may be with reference to the cell or the biotic component in virus on the capture complex.Wherein, in b step
In can by the sample be placed in the container it is front by the sample with it is described it is compound capture body contact, it is also possible to by the sample
Product contact the sample with the compound capture body after being placed in the container.Wherein, the sample is made by the mode such as shaking up
Product are fully contacted with the compound capture body, the sample is more specifically bound with the compound capture body.Wherein, exist
Also include the step of adding buffer in the sample before the sample is contacted with the compound capture body.Wherein, the above
Step is carried out under suitable temperature conditionss, for example, when separating and detecting that zooblast is particularly human cell, keeps temperature
Degree is at 30-40 degree Celsius.
On the other hand, the present embodiments relate to the detection method of micro-example amplifying nucleic acid, including：A. compound capture is obtained
Body, the compound capture body comprising magnetic core and be coated in the magnetic core outward and antibody coupling shell, the antibody
There is specific affinity to the cells in sample or the defined epitope in virus；B. by the sample be placed in a container with
And contact the sample with the compound capture body；C. apply magnetic field to the container, be adsorbed onto the capture complex
On the container inner wall, waste liquid is removed；D. add rinsing liquid to it is described capture complex wash, obtain capture complex-
Nucleic conjugate or capture complex-viral conjugates, remove waste liquid；E. PCR amplification procedures are performed, it is compound to the capture
Nucleic acid in body-cell conjugates or capture complex-viral conjugates is expanded or detected, the capture complex is to institute
State nucleic acid amplification or detection process does not produce inhibitory action；F. testing result judgement is carried out.
On the other hand, the present embodiments relate to separate the device of cells in sample or virus, including：A. it is combined capture
Body, the compound capture body comprising magnetic core and be coated in the magnetic core outward and antibody coupling shell, the antibody
There is specific affinity to the cells in sample or the defined epitope in virus；B. the sample is made with the compound capture
The device of body contact, the compound capture body is combined into the cell or virus states compound capture body-cell composition or multiple
Close capture body-virus composition；C. magnetic field generation device, produces in the device that the sample is contacted with the compound capture body
Magnetic field force makes the compound capture body and other unwanted Component seperations.
Further, the present embodiments relate to separate the device with cell in detection sample or viral nucleic acid, including：
A. be combined capture body, the compound capture body comprising magnetic core and be coated in the magnetic core it is outer and antibody coupling outside
Shell, the antibody has specific affinity to the cells in sample or the defined epitope in virus；B. make the sample with
The device of the compound capture body contact；C. magnetic field generation device；D. the cells in sample or the nucleic acid in virus are expanded
Device；E. the device of the cells in sample or the nucleic acid in virus is detected.
The present embodiments relate to method and apparatus to can be used for zooblast, plant cell, bacterial cell, funguses thin
Muscle cell, neurocyte, epithelial cell, immunocyte, leukocyte in born of the same parents or reconstitution cell, such as zooblast etc..This
The method and apparatus of inventive embodiments can be used for plant viruses, animal viruss, phage.
The present embodiments relate to the sample of method and apparatus detection can be special from plant, animal, culture fluid etc.
It is not fluid sample, such as sample from animal can be the body fluid of animal, it is the such as body fluid of people, including serum, blood plasma, complete
Blood, expectorant, cerebrospinal fluid, amniotic fluid, urine, gastrointestinal content, bone marrow fluid, saliva and perspiration etc., or tissue culture medium, cell culture
Liquid, plant sap, inoculum, virus-culturing fluid etc..
The present embodiments relate to method and apparatus can be used for detect cell or virus in biotic component include
Aminoacid, peptide, protein, lipid, monosaccharide, oligosaccharide, carbohydrate, nucleoside, nucleotide, oligonucleotide, nucleic acid, nucleic acid, dimension
Raw element and its complex.
The present embodiments relate to compound capture body magnetic core 1 include selected from paramagnetic meterial, ferrum magnetizable material and
The magnetisable material of ferrous magnetizable material, such as ferroso-ferric oxide；The capture that the method and apparatus of the embodiment of the present invention is related to is combined
The shell of body can be suitable various capsulating materials, antibody 2 is combined on shell 3, and antibody 2 is to be separated for institute or detects
Cell or viral species are selected, and the defined epitope in 2 pairs of cells in sample of antibody or virus has specificity affine
Power, preferably can specific recognition cell or virus surface expressed in abundance specific antigen antibody, such as it is thin in vain for the mankind
Born of the same parents select CD45 specific antibodies.
The rinsing liquid is rinsing liquid suitable in prior art, such as isotonic normal saline, PBS etc..
Can used in the sample of different scales the present embodiments relate to method and apparatus, it is particularly a small amount of and micro-
Amount sample, and the present embodiments relate to method and apparatus be easy to automatization and miniaturization.
The present embodiments relate to nucleic acid detection method in, by reagent needed for PCR (comprising buffer, Taq polymerase,
DNTPs, primer, water etc.) add capture complex-nucleic conjugate or capture in complex-viral conjugates, catch without the need for separating
Complex and cell or virus are obtained, PCR programs are directly entered, under hypisotonic solution and high temperature action, cell or virus will cracking
And nucleic acid is discharged as the template of PCR, capture complex does not produce inhibitory action to PCR amplifications.
The present embodiments relate to nucleic acid amplification and detecting step in, can PCR amplification after use suitable method
Directly nucleic acid is detected and analyzed, such as gel electrophoresiss, nucleic acid sequencing, making nucleic acid molecular hybridization, it is also possible to fixed using fluorescence
Amount PCR method is expanded and detected to the cell or viral nucleic acid, and the capture complex is to the amplification and detects equal
Unrestraint is acted on.
Embodiment 1. separates leukocyte and detection nucleic acid from people's whole blood
(1) CD45 specific antibodies are coupled in the compound body case of capture；
(2) 3 microlitres of people's finger tip blood samples are taken to be placed in container, under 37 degrees Celsius, 100 microlitres of HEPES buffer solutions is added
(pH7.4) capture complex, is added, is rocked, capture complex specific binding leukocyte forms capture complex-leukocyte knot
Compound, complex is captured on container inner wall using Magnet in container outer wall absorption, removes waste liquid, adds rinsing liquid multiple to capture
Zoarium-leukocyte conjugate is washed, and removes waste liquid；
(3) by reagent needed for PCR (comprising buffer, Taq polymerase, dNTPs, primer, water etc.) add capture complex-
In leukocyte conjugate, quantitative fluorescent PCR program is directly entered, under hypisotonic solution and high temperature action, leukocyte will be cracked simultaneously
Nucleic acid is discharged as the template of PCR, is expanded using the method dialogue nucleus such as quantitative fluorescent PCR, ARMS-PCR, LAMP
Increase and detect；
Embodiment 2. is from separating mouse thymic epithelial cell in thymic tissue culture fluid and detection nucleic acid
(1) by mAb RE-4D8 antibody couplings to the compound body case of capture；
(2) 3 microlitres of thymic tissue culture fluid samples are taken to mix with capture complex, in being placed in container, 100 microlitres is added
Tris-EDTA buffer (pH7.6), mixture turbine mixer is gently mixed 15 seconds, and 3 points are stood under 37 degrees Celsius
Clock, capture complex specific binding Mouse thymic epitheliai cells form capture complex-epithelial cell conjugate, using Magnet
In container outer wall absorption capture complex on container inner wall, waste liquid is removed, add PBS (pH7.4) compound to capture
Body-epithelial cell conjugate is washed, and removes waste liquid；
(3) by reagent needed for PCR (comprising buffer, Taq polymerase, dNTPs, primer, water etc.) add capture complex-
In epithelial cell conjugate, PCR programs are directly entered, under hypisotonic solution and high temperature action, Mouse thymic epitheliai cells will split
Nucleic acid is solved and discharged as the template of PCR, Mouse thymic epitheliai cells nucleic acid amplification product is obtained；
(4) after PCR terminates, nucleic acid amplification product is detected and analyzed using gel electrophoresiss.
Embodiment 3. separates M13 phagies and detection nucleic acid from inoculum
(1) by anti-M13 antibody couplings to the compound body case of capture；
(2) take 100 microlitres of inoculum samples to be placed in container, with Hanks liquid by antigen diluent to 10 μ g/ml, plus
Enter to capture complex mixing, mixture turbine mixer be gently mixed 15 seconds, be incubated 3 minutes under 37 degrees celsius,
Capture complex specific binding M13 phagies form capture complex-phage conjugate, are inhaled in container outer wall using Magnet
Attached capture complex removes waste liquid on container inner wall, adds PBS (pH7.4) to combine to capturing complex-phage
Thing is washed, and removes waste liquid；
(3) by reagent needed for PCR (comprising buffer, Taq polymerase, dNTPs, primer, water etc.) add capture complex-
In phage conjugate, PCR programs are directly entered, under hypisotonic solution and high temperature action, M13 phagies will be cracked and discharged
Nucleic acid obtains M13 bacteriophage nucleic acid amplified productions as the template of PCR；
(4) after PCR terminates, nucleic acid amplification product is detected and analyzed using gel electrophoresiss.
Whether there is H5N1 influenza A viruss in the detection blood plasma of embodiment 4.
(1) influenza A viruss nucleoprotein monoclonal antibody after purification is coupled in the compound body case of capture；
(2) 3 microlitres of plasma samples are taken to be placed in container, 100 microlitres of PBSs (pH6.8) are added, adds capture compound
Body mixes, and mixture turbine mixer is gently mixed 15 seconds, is incubated 30 minutes under 37 degrees celsius, if depositing in blood plasma
In H5N1 influenza A viruss, then capture complex specific binding H5N1 influenza A viruss and form capture complex-virus
Conjugate, complex is captured on container inner wall using Magnet in container outer wall absorption, removes waste liquid, adds PBS
(pH6.8) wash to capturing complex-viral conjugates, remove waste liquid；
(3) by reagent needed for PCR (comprising buffer, Taq polymerase, dNTPs, primer, water etc.) add capture complex-
In viral conjugates, PCR programs are directly entered, under hypisotonic solution and high temperature action, if there is H5N1 influenza As in blood plasma
Virus, then H5N1 influenza A viruss will crack and discharge nucleic acid as the template of PCR, obtain H5N1 influenza A viruss cores
Sour amplified production；
(4) after PCR terminates, nucleic acid amplification product is detected and analyzed using making nucleic acid molecular hybridization method.
Description to the various embodiments of the present invention above is supplied to those skilled in the art with the purpose for describing.It is not
Be intended to exhaustion or be not intended to limit the invention to single disclosed embodiment.As described above, the present invention's is various
Substitute and change will be apparent for above-mentioned technology one of ordinary skill in the art.Therefore, although specifically beg for
The embodiment of some alternatives has been discussed, but other embodiment will be apparent, or those skilled in the art are relative
Easily draw.It is contemplated that all replacements of the invention, modification and the change that had discussed including here, and fall
Other embodiment in the spirit and scope of above-mentioned application.
Although depicting the present invention by embodiment, it will be appreciated by the skilled addressee that the present invention has many deformations
With change without deviating from spirit of the invention, it is desirable to which appended claim includes these deformations and changes without deviating from the present invention
Spirit.
Claims (9)
1. the detection method of micro-example amplifying nucleic acid, including：A. compound capture body is obtained, the compound capture body includes magnetic core
The heart and the shell being coated in outside the magnetic core, the shell and antibody coupling, the antibody to the cells in sample or
Defined epitope in virus has specific affinity；B. by the sample be placed in a container and by the sample with it is described
Compound capture body contact, makes capture complex combine with cell or virus；C. apply magnetic field to the container, make the capture multiple
Zoarium is adsorbed onto on the container inner wall, removes waste liquid；D. add rinsing liquid to wash the capture complex, caught
Complex-cell conjugates or capture complex-viral conjugates are obtained, waste liquid is removed；E. amplification process is performed, to described
Nucleic acid in capture complex-cell conjugates or capture complex-viral conjugates is expanded or detected, the capture is multiple
Zoarium does not produce inhibitory action to the nucleic acid amplification or detection process；F. testing result judgement is carried out.
2. method as claimed in claim 1, the method for the nucleic acid amplification is selected from polymerase chain reaction, reverse transcription PCR, nido
PCR, multiplex PCR, semiquantitive PCR, real-time fluorescence quantitative PCR, In situPCR, ligase chain reaction, randomly amplified polymorphic DNA,
Ring mediated isothermal amplification, Nucleic acid sequence amplification, recombinant PCR, anchor PCR, asymmetric PCR, inverse PCR or immuno-PCR
In one or more.
3. method as claimed in claim 1 or 2, is additionally included in before the sample is contacted with the compound capture body to the sample
The step of buffer being added in product.
4. method as claimed in claim 1, the method that the testing result judges is selected from gel electrophoresiss, nucleic acid sequencing, nucleic acid point
One or more in son hybridization.
5. method as claimed in claim 1 or 2, the sample is selected from lymph fluid, tear, perspiration, urine, saliva, whole blood, essence
In liquid, cerebrospinal fluid, lung cavity liquid, peritoneal fluid, the liquid in joint, amniotic fluid, interstitial fluid, tissue culture medium or cell culture fluid
One or more.
6. method as claimed in claim 1 or 2, the cell is selected from human body cell, zooblast, plant cell, antibacterial, funguses
One or more in cell or reconstitution cell.
7. method as claimed in claim 6, the human body cell selected from Colonic exfoliative cells, exuviation cell, phagocyte,
One or more in leukocyte, stem cell, neurocyte.
8. method as claimed in claim 1 or 2, the antibody is selected from CD45 specific antibodies, E.coli antibody, mAb RE-
One or more in 4D8, anti-M13 antibody, influenza A viruss nucleoprotein monoclonal antibody.
9. the device of micro-example amplifying nucleic acid is detected, including：A. be combined capture body, the compound capture body comprising magnetic core and
The shell being coated in outside the magnetic core, the shell and antibody coupling, the antibody is to the cells in sample or virus
On defined epitope there is specific affinity；B. the device for making the sample contact with the compound capture body；C. is produced from magnetic field
Generating apparatus；D. the device of the cells in sample or the nucleic acid in virus is expanded；E. detect in the cells in sample or virus
Nucleic acid device.
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