CN104059849A - Device and method used for rapid extraction of nucleic acid - Google Patents
Device and method used for rapid extraction of nucleic acid Download PDFInfo
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- CN104059849A CN104059849A CN201410333236.7A CN201410333236A CN104059849A CN 104059849 A CN104059849 A CN 104059849A CN 201410333236 A CN201410333236 A CN 201410333236A CN 104059849 A CN104059849 A CN 104059849A
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1017—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by filtration, e.g. using filters, frits, membranes
Abstract
The invention relates to a device and method used for rapid extraction of nucleic acid which are mainly used for rapidly extracting nucleic acid (including RNA and DNA) from each clinical sample. The device mainly comprises an injector, a hollow adaptor, a nucleic acid adsorption membrane tube and a sampling probe, after the steps of splitting a clinical sample, sucking by virtue of an injector, adsorbing nucleic acid by virtue of a nucleic acid adsorption film, rinsing the nucleic acid adsorption film with rinsing liquid and eluting nucleic acid with eluant are carried out, a pure nucleic acid solution can be obtained to be used in a downstream experiment.
Description
Technical field
The present invention relates to biological technical field, be specifically related to a kind of can be from various clinical samples the device and method of rapid extraction nucleic acid.
Background technology
(1) nucleic acid extraction
Nucleic acid extraction has become most important, the most basic operation in molecular biology experiment technology, but the extraction scheme of nucleic acid is many at present, and complicated and simple differing.The successful extraction that completes whole dependence nucleic acid of the related experiment such as molecular diagnosis, nucleic acid sequencing, functional genome, oneself can realize whole automatizations by warp traditional operating process, but these expensive instruments can not be promoted the use of on a large scale, China is still developing country at present, vast different medical unit can not be equipped with complicated instrument, therefore, provide a kind of nucleic acid-extracting apparatus of good economy performance necessary.
Nucleic acid comprises DNA, two kinds of molecules of RNA, in cell, be all to exist with the state of protein bound, the key step of nucleic acid extraction is: one, lysing cell, the biomacromolecules such as the protein that removal is combined with nucleic acid and polysaccharide, lipid, remove other unwanted nucleic acid molecule, during as extraction DNA molecular, should remove RNA, vice versa, if extraction RNA and DNA extract altogether, do not need to remove any; Two, precipitate nucleic acids or absorption nucleic acid; Three, purification of nucleic acid, removes salt, the impurity such as organic agent; Four, wash-out or dissolving nucleic acid, utilize elutriant or lysate by nucleic acid wash-out or dissolving.
(2) clinical sample pre-treatment
Because processing and the preservation of sample are larger on the impact of DNA and RNA (especially RNA), therefore in the time of nucleic acid determination, the processing of sample and suitable preservation are extremely important to the accurate and effective of measurement result.Clinical common sample has serum (slurry), whole blood, secretory product, cotton swab, fester, body fluid, flesh tissue, paraffin section etc., and the treatment process of these clinical samples is had nothing in common with each other.
1. serum (slurry) sample
Add appropriate erythrocyte cracked liquid (rupture of membranes liquid), the red corpuscle in cracking whole blood; Add appropriate nucleus again and split liquid (broken karyolymph), the nucleus in cracking white corpuscle, discharges nuclear dna; And then add the stain removers such as SDS (sodium lauryl sulphate), dissolve lipoprotein, depolymerization nucleoprotein; SDS can form mixture with protein, makes protein denaturation precipitation, but must remove in SDS nucleic acid extraction process afterwards.
2. sputum specimen
Phlegm belongs to secretory product, be commonly used for clinically mycobacterium tuberculosis dna and measure sample, owing to containing a large amount of Saliva Orthanas and impurity in sputum specimen, therefore in the time of nucleic acid extraction, must carry out pre-treatment to sample, with 4%NaOH liquefaction, remove Saliva Orthana, then with boiling or the classical approach extraction DNA such as proteolytic enzyme, phenol/chloroform.
3. cotton swab
While detecting venereal disease pathogenic agent by PCR method (chlamydozoan, mycoplasma, gonococcus, treponema pallidum, human papillomavirus etc.), clinical samples is generally cotton swab.Specimen sampling is crucial, and the pathogenic infection epithelial cells such as chlamydozoan, therefore draw materials and must get cell.Concrete grammar:
Add 1ml stroke-physiological saline solution, fully concussion mixes, 12000rpm, centrifugal 5 minutes
Get urethral secretions or cervical secretions (being gathered by medical personnel) with cotton swab, put in sterile test tube or EP pipe and abandon supernatant, precipitation adds stroke-physiological saline solution 1ml, beats 12000rpm, centrifugal 5 minutes
4. body fluid
Humoral specimen, comprises hydrothorax, ascites, cerebrospinal fluid, urine etc., can be directly centrifugal, and taking precipitate extracts nucleic acid.Courageous and upright ascites pleural fluid, can use erythrocyte cracked liquid processing:
5. fester
Thickness fester: with sputum specimen processing, first with 4%NaOH liquefaction, then centrifuging and taking precipitation is extracted DNA.
Water sample fester: directly centrifugal, precipitation with physiological saline wash 2 ?after 3 times for DNA extraction.
6. tissue
Tissue has flesh tissue and paraffin section.The treatment step of flesh tissue: first wash secondary with physiological saline, then smashed or shred to pieces (with eye scissors), extract nucleic acid after adding protease K digesting; Paraffin section, for nucleic acid extraction, needs first to dewax with dimethylbenzene, then with carrying out DNA extraction after protease K digesting.
(3) nucleic acid-extracting apparatus
CN101684463A proposes a kind of trace clinical sample nucleic acid rapid extraction device, by sharp-crested suction pipe, strainer with film and suction unit (syringe or plastic blister) three part compositions, it has aspirated nucleic acid absorption by three times, clean and elution process, the fast and convenient nucleic acid that extracts from sample of energy, so blemish in an otherwise perfect thing is, sharp-crested suction pipe, strainer with film and suction unit three part array modes single and for fixed installation, (Fig. 2 of CN101684463A while making suction unit by plastic blister, Fig. 3) suction for the third time carries out that nucleic acid wash-out is actual can not complete that (waste liquid of front twice suction cannot be discharged, elutriant with nucleic acid is mixed in waste liquid), and use syringe while making suction unit, enter syringe with the elutriant of nucleic acid and not only can be infected with syringe formation biologic garbage, also can cause trace sample amplifying nucleic acid remarks emitter to be infected with and lose cause detect inaccurate.
Summary of the invention
For overcoming problems of the prior art, the object of the invention be to provide a kind of simple, practical, efficient for device and a kind of nucleic acid rapid extracting method of nucleic acid rapid extraction.
Device for nucleic acid rapid extraction of the present invention, comprise syringe, adsorption film and sample needle, also comprise the interconnecting device of a hollow, described adsorption film is arranged on the bottom in an adsorption film pipe, and interconnecting device is connected in the middle of syringe and adsorption film pipe and syringe, interconnecting device and adsorption film Guan Ye road are communicated with.
Adsorption film pipe is the body of both ends open, the opening end that one end is this body, and the other end is fixed an osculum pipe connecting, and adsorption film is arranged on this osculum pipe connecting one end; Adsorption film pipe opening end size is mated with the outside dimension of interconnecting device.
Syringe comprises cylindrical shell, be placed in the tubulose Connection Block of barrel front end and be placed in cylindrical shell can push-and-pull push rod, the external diameter of described osculum pipe connecting equates with the Connection Block outside dimension of syringe, described interconnecting device is established intermediate throughholes, and intermediate throughholes size is mated with the Connection Block outside dimension of syringe.
Described sample needle comprises needle stand and syringe needle, and the external diameter of described osculum pipe connecting mates with the needle stand internal diameter of described sample needle.
Interconnecting device one end is communicated with syringe, and the other end is communicated with the osculum pipe connecting of adsorption film pipe, forms the drawing liquid mode of this device.
Or interconnecting device one end is communicated with syringe, the other end is socketed in the opening end of adsorption film pipe, and the osculum pipe connecting of adsorption film pipe is communicated with sample needle, forms the liquid mode that pushes away of this device.
Described interconnecting device profile is round table-like, and the microcephaly of round table-like interconnecting device holds outside dimension to mate with the opening end of adsorption film pipe, and this opening end is held socket from the microcephaly of round table-like interconnecting device.
Another object of the present invention is to provide a kind of method of nucleic acid rapid extraction.
The method of nucleic acid rapid extraction provided by the invention, use aforementioned nucleic acid rapid extraction device, after the steps such as sample cracking processing, adsorption film absorption sample amplifying nucleic acid, rinsing liquid rinsing adsorption film removal of impurities and elutriant wash-out nucleic acid, obtain pure nucleic acid solution.
The method can be " two take out one pushes away " method: " one takes out " is adsorbed on filter membrane the nucleic acid in lysate; " two take out " make rinsing liquid by filter membrane with clean impurity; " one pushes away " by the nucleic acid component wash-out being adsorbed on filter membrane, thereby has completed the extraction of biological specimen amplifying nucleic acid composition; Specifically comprise the following steps:
A) reagent indicates packing:
Each person-portion such as the lysate that may relate in nucleic acid extraction, RNA adsorption liquid, RNA enzyme inhibitors, 95% alcohol, 75% alcohol, rinsing liquid, elutriant is divided and is filled to 1.5EP pipe, and each pipe is indicated;
B) clinical sample is added in lysate pipe and (extract if relate to RNA, then add RNA adsorption liquid and RNA enzyme inhibitors), and mix;
C) one inhales: the drawing liquid form that uses nucleic acid-extracting apparatus, from sample cracking mixed solution, aspirate with the opening end of adsorption film pipe, the suction that utilizes syringe pump to produce, when nucleic acid component in sample is flowed through adsorption film because the specific adsorption of film is adsorbed on adsorption film, filtrate being has the waste liquid of the impurity such as cell debris and protein, and waste liquid enters syringe cavity and waste liquid only passes through this adsorption film once;
D) two inhale: the drawing liquid form that uses nucleic acid-extracting apparatus, from rinsing liquid pipe, aspirate with the opening end of adsorption film pipe, the suction that utilizes syringe pump to produce, clean adsorption film with rinsing liquid, cleaning remains in protein or other composition on adsorption film, and filtrate enters syringe cavity and this filtrate is only passed through adsorption film once;
E) one push away: the syringe of changing another cleaning, first suck elutriant, then use nucleic acid-extracting apparatus to push away liquid form (reverse by adsorption film pipe, make adsorption film front, opening end socket interconnecting device one end of adsorption film pipe, the osculum pipe connecting of adsorption film pipe connects the needle stand of sample needle), elutriant in syringe cavity is joined in adsorption film pipe, utilize pressure that syringe pushing produces by eluent stream through adsorption film, and the nucleic acid component wash-out being adsorbed on film is pushed to follow-up nucleic acid collection container or proofing unit, complete the leaching process of sample amplifying nucleic acid.
The method also can be " three push away " method: " one pushes away " is adsorbed on filter membrane the nucleic acid in lysate; " two push away " makes rinsing liquid clean impurity by filter membrane; " three push away " by the nucleic acid component wash-out being adsorbed on filter membrane, thereby has completed the extraction of biological specimen amplifying nucleic acid composition; Specifically comprise the following steps:
A) reagent indicates packing:
Each person-portion such as the lysate that may relate in nucleic acid extraction, RNA adsorption liquid, RNA enzyme inhibitors, 95% alcohol, 75% alcohol, rinsing liquid, elutriant is divided and is filled to 1.5EP pipe, and each pipe is indicated to (using different letters and color differentiating);
B) clinical sample is added in lysate pipe and (extract if relate to RNA, then add RNA adsorption liquid and RNA enzyme inhibitors), and mix.
C) first suck sample cracking mixed solution with syringe, (adsorption film pipe is reverse for the liquid form that pushes away of use nucleic acid-extracting apparatus, do not establish sample needle), the pressure that utilizes syringe pushing to produce makes the cracking mixed solution filtering membrane of flowing through, when nucleic acid component in sample is flowed through adsorption film because the specific adsorption of film is adsorbed on adsorption film, filtrate being has the waste liquid of the impurity such as cell debris and protein, and waste liquid is discharged from and waste liquid only passes through adsorption film once;
D) suck again rinsing liquid with syringe, (adsorption film pipe is reverse for the liquid form that pushes away of use nucleic acid-extracting apparatus, do not establish sample needle), utilize pressure that syringe pushing produces that rinsing liquid is joined and in adsorption tube, cleans adsorption film, clear protein or other composition remaining on adsorption film, filtrate is discharged from and this filtrate is only passed through adsorption film once;
E) syringe of replacing another cleaning sucks elutriant, (adsorption film pipe is reverse for the liquid form that pushes away of use nucleic acid-extracting apparatus, add sample needle), elutriant in syringe cavity is joined in adsorption film pipe, utilize pressure that syringe pushing produces by eluent stream through adsorption film, and the nucleic acid component wash-out being adsorbed on film is pushed to follow-up nucleic acid collection container or proofing unit, complete the leaching process of sample amplifying nucleic acid.
Adopt above scheme, nucleic acid-extracting apparatus of the present invention adopts bulk-breaking mode, by the assembling use of arranging in pairs or groups, is skillfully constructed, simple in structure, practical.Wherein, interconnecting device two ends can connect different parts simultaneously, and adsorption film pipe detachably and oppositely assembles, and are used in conjunction with hollow and the identical interconnecting device of size, complete the collocation of nucleic acid extraction from absorption washing to wash-out different steps, make nucleic acid extraction simple, quick, efficient.Meanwhile, the present invention utilizes syringe to replace high speed centrifuge, utilizes extraction element just can in liquid phase heterogeneous system, reach the effect of separation.
The present invention can be used as the front simple and direct means of one of biological specimen being carried out to nucleic acid extraction of nucleic acid amplification, the sample purity extracting enough meets the requirement of next step nucleic acid amplification, simultaneously without any need for instrument, operational safety and simple, disposable use, thereby can be widely used in pathogenic agent or host's nucleic acid extraction, for clinical detection and diagnosis.Extraction element of the present invention is with low cost, while simple to operate is again without any need for specific apparatus, operating time is short, approximately only need 10 ?15 minutes, thereby the present invention can be widely used in the relatively poor basic hospital of some experiment conditions and economically less developed region or country; Meanwhile, also can meet well the demand, particularly emergency room of high-level hospital and the nucleic acid quick diagnosis of bedside.
Brief description of the drawings
Fig. 1 is the device exploded view of the present invention for nucleic acid rapid extraction;
Decomposition and constitutional diagram when Fig. 2 device is exercised pumping function;
Decomposition and constitutional diagram when Fig. 3 device is exercised pushing function;
The Salmonellas DNA pcr amplification detected result that Fig. 4 example 1 of the present invention obtains;
The Trichinella spiralis DNA cloning detection of nucleic acids result that Fig. 5 embodiment of the present invention 2 obtains;
The H1N1RNA RT that Fig. 6 embodiment of the present invention 3 obtains ?LAMP augmentation detection result.
Embodiment
The invention provides for the device of nucleic acid rapid extraction and the method for nucleic acid extraction, for from various clinical sample rapid extraction nucleic acid (comprising RNA and DNA).
Device for nucleic acid rapid extraction provided by the invention referring to figure 1 ?shown in Fig. 3, mainly comprise syringe 1, adsorption film pipe 2, interconnecting device 3 and 4 four parts of sample needle, wherein:
Syringe 1 is common tubular disposable syringe or the allied equipment that can exercise syringe function, it has cylindrical shell 11, be placed in cylindrical shell 11 front ends for connect the tubulose Connection Block 12 of syringe needle and be placed in cylindrical shell 11 can push-and-pull push rod 13; Sample needle 4 is common injection needles, comprises needle stand 41 and syringe needle 42.
Cylinder or round platform that interconnecting device 3 is hollow, its intermediate throughholes 31 sizes are mated with syringe 1 tubulose Connection Block 12 outside dimensions, and this tubulose Connection Block 12 can be inserted in interconnecting device 3 intermediate throughholes 31; Separately, interconnecting device 3 appearances are cylindric or round table-like, and its outside dimension mates with adsorption film pipe 2, below describe in detail; Interconnecting device 3 can have elastic material and make as rubber, plastics.
The body that adsorption film pipe 2 is both ends open, available plastic material is made, its one end is the opening end 21 of this body, the other end is fixed an osculum pipe connecting 22, adsorption film pipe 2 inner bottom parts (pipe connecting 22 one end are called the end) are established adsorption film 23 (commercial goods, as pellosil Silica membrane); The both ends open of adsorption film pipe 2 all can be connected with interconnecting device 3, and wherein opening end 21 sizes are mated with the outside dimension of cylindric interconnecting device 3, or holds outside dimension to mate with the microcephaly of round table-like interconnecting device so that interconnecting device 3 can be socketed with opening end 21; Osculum pipe connecting 22 external diameters of adsorption film pipe 2 are identical with syringe 1 Connection Block 12, both mated with needle stand 41 internal diameters of sample needle 4, again with interconnecting device 3 intermediate throughholes 31 size match, make this osculum pipe connecting 22 can be inserted into the interior connection sample needle 4 of needle stand 41, can be inserted into again connection adapter 3 in intermediate throughholes 31.
In the present invention, while extracting solution, connect the Connection Block 12 of syringe 1 with interconnecting device 3 one end, the other end connects the osculum pipe connecting 22 of adsorption film pipe 2, make adsorption film 23 rear (in the present invention with syringe user to definition before and after, immediate shipment syringe needle one end is defined as " front ", and when push rod extracts, direction of motion is defined as " afterwards "), combine the drawing liquid form (as shown in Figure 2) of nucleic acid rapid extraction device.While releasing solution, connect the Connection Block 12 of syringe 1 with interconnecting device 3 one end, the other end stretches into the opening end 21 of adsorption film pipe 2, and the osculum pipe connecting 22 of adsorption film pipe 2 connects the needle stand 41 of sample needle 4 again, combines the liquid form that pushes away (as shown in Figure 3) of nucleic acid rapid extraction device.
Use the device of the present invention for nucleic acid rapid extraction, after the steps such as clinical sample cracking processing, syringe pump lysate, nucleic acid adsorption film absorption nucleic acid, rinsing liquid rinsing nucleic acid adsorption film, elutriant wash-out nucleic acid, can obtain pure nucleic acid solution and test for downstream.
The ultimate principle of nucleic acid extraction of the present invention, that adsorption film (also can be described as filter membrane) in adsorption tube has special adsorption to nucleic acid, by the nucleic acid absorption in sample and cracking mixed solution, then purify through rinsing the object that reaches purification, finally from adsorption film, wash-out out, ensures on a small quantity or the maximum yield of high value sample amplifying nucleic acid with separating of sample to realize nucleic acid again.Under normal circumstances, the nucleic acid extraction after cracking need to be in liquid phase heterogeneous system, utilizes the centrifugal force of whizzer to reach the separation of liquid liquid, the effect of liquid-solid separation.But apparatus of the present invention only need " two take out once pushing away " or " three push away " can complete all operations." two take out one pushes away " is: " one takes out ": the nucleic acid in lysate is adsorbed on filter membrane; " two take out ": make rinsing liquid complete the object of cleaning impurity by filter membrane; " one pushes away ": by the nucleic acid component wash-out being adsorbed on filter membrane, thereby completed the extraction of biological specimen amplifying nucleic acid composition." three push away ": " one pushes away ": the nucleic acid in lysate is adsorbed on filter membrane; " two push away ": make rinsing liquid complete the object of cleaning impurity by filter membrane; " three push away ": by the nucleic acid component wash-out being adsorbed on filter membrane, thereby completed the extraction of biological specimen amplifying nucleic acid composition.
For this reason, method of getting nucleic acid provided by the invention is used above nucleic acid-extracting apparatus, and a kind of method is called as " two take out one pushes away " method, comprises the following steps:
A) reagent indicates packing:
Each person-portion such as the lysate that may relate in nucleic acid extraction, RNA adsorption liquid, RNA enzyme inhibitors, 95% alcohol, 75% alcohol, rinsing liquid, elutriant is divided and is filled to 1.5EP pipe, and each pipe is indicated to (using different letters and color differentiating);
B) clinical sample is added in lysate pipe and (extract if relate to RNA, then add RNA adsorption liquid and RNA enzyme inhibitors), and mix;
C) one inhales: the drawing liquid form that uses nucleic acid-extracting apparatus, from sample cracking mixed solution, aspirate with the opening end of adsorption film pipe, the suction that utilizes syringe pump to produce, when nucleic acid component in sample is flowed through adsorption film because the specific adsorption of film is adsorbed on adsorption film, filtrate being has the waste liquid of the impurity such as cell debris and protein, and waste liquid enters syringe cavity and waste liquid only passes through this adsorption film once;
D) two inhale: the drawing liquid form that uses nucleic acid-extracting apparatus, from rinsing liquid pipe, aspirate with the opening end of adsorption film pipe, the suction that utilizes syringe pump to produce, clean adsorption film with rinsing liquid, cleaning remains in protein or other composition on adsorption film, and filtrate enters syringe cavity and this filtrate is only passed through adsorption film once;
E) one push away: the syringe of changing another cleaning, first suck elutriant, then use nucleic acid-extracting apparatus to push away liquid form (reverse by adsorption film pipe, make adsorption film front, opening end socket interconnecting device one end of adsorption film pipe, the osculum pipe connecting of adsorption film pipe connects the needle stand of sample needle), elutriant in syringe cavity is joined in adsorption film pipe, utilize pressure that syringe pushing produces by eluent stream through adsorption film, and the nucleic acid component wash-out being adsorbed on film is pushed to follow-up nucleic acid collection container or proofing unit, complete the leaching process of sample amplifying nucleic acid.What so obtain is the aqueous solution nucleate that does not contain other impurity, can be used for nucleic acid amplification.
Method of getting nucleic acid provided by the invention is used above nucleic acid-extracting apparatus, and another kind of method is called as " three push away " method, comprises the following steps:
A) reagent indicates packing:
Each person-portion such as the lysate that may relate in nucleic acid extraction, RNA adsorption liquid, RNA enzyme inhibitors, 95% alcohol, 75% alcohol, rinsing liquid, elutriant is divided and is filled to 1.5EP pipe, and each pipe is indicated to (using different letters and color differentiating);
B) clinical sample is added in lysate pipe and (extract if relate to RNA, then add RNA adsorption liquid and RNA enzyme inhibitors), and mix.
C) first suck sample cracking mixed solution with syringe, (adsorption film pipe is reverse for the liquid form that pushes away of use nucleic acid-extracting apparatus, do not establish sample needle), the pressure that utilizes syringe pushing to produce makes the cracking mixed solution filtering membrane of flowing through, when nucleic acid component in sample is flowed through adsorption film because the specific adsorption of film is adsorbed on adsorption film, filtrate being has the waste liquid of the impurity such as cell debris and protein, and waste liquid is discharged from and waste liquid only passes through adsorption film once;
D) suck again rinsing liquid with syringe, (adsorption film pipe is reverse for the liquid form that pushes away of use nucleic acid-extracting apparatus, do not establish sample needle), utilize pressure that syringe pushing produces that rinsing liquid is joined and in adsorption tube, cleans adsorption film, clear protein or other composition remaining on adsorption film, filtrate is discharged from and this filtrate is only passed through adsorption film once;
E) syringe of replacing another cleaning sucks elutriant, (adsorption film pipe is reverse for the liquid form that pushes away of use nucleic acid-extracting apparatus, add sample needle), elutriant in syringe cavity is joined in adsorption film pipe, utilize pressure that syringe pushing produces by eluent stream through adsorption film, and the nucleic acid component wash-out being adsorbed on film is pushed to follow-up nucleic acid collection container or proofing unit, complete the leaching process of sample amplifying nucleic acid.What so obtain is the aqueous solution nucleate that does not contain other impurity, can be used for nucleic acid amplification.
The nucleic acid extracting through the present invention can be applied to the downstream experiment that microorganism detection, detection in Gene Mutation, single nucleotide polymorphism detection etc. relate to nucleic acid.Nucleic acid is for can be DNA or RNA.
The solution that isothiocyanic acid flesh solution or the Chelex lysate etc. commonly used clinically can lysing cell for the present invention's lysate; Rinsing liquid is the scavenging solution of PH<7, as 70% alcohol; Elutriant is Tris-HCL or the distilled water of 10mM; Adsorption film is the various films that can adsorb nucleic acid, as pellosil (Silica membrane).All commercially available acquisitions of agents useful for same and material in the present invention.
With example, application of the present invention is described below.
Example 1: the extraction of Salmonellas DNA
This example, with the example that is extracted as of Salmonellas DNA, verifies that whether nucleic acid-extracting apparatus of the present invention and method be effective.Totally 4 parts, this Cass collection sample (1 negative contrast, 2 ?4 be Salmonellas), proceeds as follows respectively:
1. cracking
Get the bacterium liquid of the Salmonellas incubated overnight of 500 microlitres, add the lysate (composition: NaOH, 25mmol of equivalent; Tris ?Hcl, 10mmol; Triton X ?100,1%; NP ?40,1%; EDTA, 0.1mMol; Chelex ?100,2%.); Mix rear room temperature and leave standstill 5 minutes, use the drawing liquid form of nucleic acid-extracting apparatus, mixed solution is pumped to adsorption film pipe, make mixed solution through adsorption film, do not enter syringe cavity containing the waste liquid of nucleic acid.
2. clean
The drawing liquid form that uses nucleic acid-extracting apparatus, is drawn into rinsing liquid in adsorption film pipe, and the same step operation makes rinsing liquid clean the impurity on adsorption film, and waste liquid enters into syringe cavity.
3.DNA wash-out
Change a clean syringe, 120 microlitre elutriants are sucked into syringe cavity, use the liquid form that pushes away of nucleic acid-extracting apparatus, push with syringe the power producing elutriant is pushed into adsorption film pipe, eluent stream is through adsorption film, and push to a new centrifuge tube by sample needle together with elutriant being adsorbed on nucleic acid component on film, for subsequent use.
4. nucleic acid amplification
Application salmonella PCR amplifing reagent (2 × Tap MIX test kit, TIANGEN Biotech (Beijing) Co., Ltd.), the DNA that amplification extracts respectively, concrete reaction is as follows: Salmonellas amplification reaction solution 20 microlitres, nucleic acid (DNA) solution 5 microlitres that extract in centrifuge tube, cumulative volume is 25 microlitres, response procedures: 95 DEG C, and circulation in 5 minutes; 94 DEG C, 30 seconds, 58 DEG C, 30 seconds, 72 DEG C, totally 35 circulations in 30 seconds; 72 DEG C, circulation in 7 minutes; 4 DEG C of preservations.
5. result detects
Amplified production detects by electrophoresis, result as accompanying drawing 4 (wherein M is NDA marker, 1 negative contrast, 2 ?4 be Salmonellas).Can find out from the result of Fig. 4, the detected result of Salmonellas sample is all positive, shows that Salmonellas DNA extraction of the present invention is very effective.
Example 2: loop-mediated isothermal amplification method detects the Trichinella spiralis in blood
This example detects Trichinella spiralis in blood as example taking loop-mediated isothermal amplification method, and the application of nucleic acid-extracting apparatus of the present invention and method is described.Detect totally 8 parts, sample (1 ?the positive infection blood sample of 6 sample, 7 ?the negative contrast of 8 sample), carry out respectively the following operation that detects:
1. cracking
Get the blood sample of 500 microlitres, add the lysate (composition: NaOH, 25mmol of equivalent; Tris ?Hcl, 10mmol; Triton X ?100,1%; NP ?40,1%; EDTA, 0.1mMol; Chelex ?100,2%.); Mix rear room temperature and leave standstill 5 minutes, use the drawing liquid form of nucleic acid-extracting apparatus, mixed solution is pumped to adsorption film pipe, make mixed solution through adsorption film, do not enter syringe cavity containing the waste liquid of nucleic acid.
2. clean
The drawing liquid form that uses nucleic acid-extracting apparatus, is drawn into rinsing liquid in adsorption film pipe, and the same step operation makes rinsing liquid clean the impurity on adsorption film, and waste liquid enters into syringe cavity.
3.DNA wash-out
Change a clean syringe, 120 microlitre elutriants are sucked into syringe cavity, use the liquid form that pushes away of nucleic acid-extracting apparatus, push with syringe the power producing elutriant is pushed into adsorption film pipe, eluent stream is through adsorption film, and push to a new centrifuge tube by sample needle together with elutriant being adsorbed on nucleic acid component on film, for subsequent use.
4. nucleic acid amplification
Application Trichinella spiralis loop-mediated isothermal amplification reagent (LAMP method DNA cloning test kit (DNA Amplification Kit), Japan (the EIKEN CHEMICAL CO. of Eiken Chemical, LTD, Tochigi, Japan) the Trichinella spiralis DNA that), amplification extracts.Concrete reaction is as follows: Trichinella spiralis amplification reaction solution 20 microlitres, and the nucleic acid extracting in centrifuge tube (DNA) solution 5 microlitres, cumulative volume is 25 microlitres, temperature of reaction and time: 65 °, react 60 minutes.
5. result detects
Amplified production is detected with real-time turbidimeter.Result is as Fig. 5.The result demonstration of Fig. 5, the positive blood sample (sample 1 ?6) infecting detection of nucleic acids result after amplification is all positive.Show that in this example extraction blood sample, Trichinella spiralis DNA can be used in detection.
Example 3: the extraction of Influenza virus H1N1 RNA and detection in throat swab
This example, taking the extraction of Influenza virus H1N1 RNA in throat swab and detection as example, illustrates that nucleic acid-extracting apparatus of the present invention and method are also suitable for extracting RNA.Detect totally 8 parts, sample (1 negative contrast, 2 ?8 be positive sample), carry out respectively extracting and detecting operation as follows:
1. cracking
Get throat swab sample 200u1, add appropriate lysate (composition: NaOH, 25mmol; Tris ?Hcl, 10mmol; Triton X ?100,1%; NP ?40,1%; EDTA, 0.1mMol; Chelex ?100,2%.), RNA adsorption liquid (composition: add equivalent 99% ethanol in lysate) and RNA enzyme inhibitors (composition: Carrier RNA), fully mix standing 5 minutes of room temperature; The drawing liquid form that uses nucleic acid-extracting apparatus, is pumped to adsorption film pipe by mixed solution, makes mixed solution through adsorption film, does not enter syringe cavity containing the waste liquid of nucleic acid.
2. clean
The drawing liquid form that uses nucleic acid-extracting apparatus, is drawn into rinsing liquid in adsorption film pipe, and the same step operation makes rinsing liquid clean the impurity on adsorption film, and waste liquid enters into syringe cavity.
3.RNA wash-out
Change a clean syringe, 120 microlitre elutriants are sucked into syringe cavity, use the liquid form that pushes away of nucleic acid-extracting apparatus, push with syringe the power producing elutriant is pushed into adsorption film pipe, eluent stream is through adsorption film, and push to a new centrifuge tube by sample needle together with elutriant being adsorbed on nucleic acid component on film, for subsequent use.
4. nucleic acid amplification
Application RT ?LAMP constant-temperature amplification reagent (LAMP method RNA amplification kit (RNA Amplification Kit), Eiken Chemical of Japan (EIKEN CHEMICAL CO., LTD, Tochigi, Japan) H1N1RNA that), amplification extracts.Concrete reaction is as follows: RT ?LAMP amplification reaction solution 20 microlitres, and the nucleic acid extracting in centrifuge tube (RNA) solution 5 microlitres, cumulative volume is 25 microlitres, temperature of reaction and time: 60 °, 60 minutes.
5. result detects
Amplification detects with real-time turbidimeter, and result is as Fig. 6 (1 negative contrast, 2 ?8 be positive sample).The result of Fig. 6 can find out, the detected result of all positive sample is all positive.Show this example can extract H1N1RNA and for detection of in.
Claims (10)
1. for the device of nucleic acid rapid extraction, comprise syringe, adsorption film and sample needle, it is characterized in that, also comprise the interconnecting device of a hollow, described adsorption film is arranged on the bottom in an adsorption film pipe, and interconnecting device is connected in the middle of syringe and adsorption film pipe and syringe, interconnecting device and adsorption film Guan Ye road are communicated with.
2. according to claim 1 for the device of nucleic acid rapid extraction, it is characterized in that, the body that adsorption film pipe is both ends open, the opening end that one end is this body, the other end is fixed an osculum pipe connecting, and adsorption film is arranged on this osculum pipe connecting one end; Adsorption film pipe opening end size is mated with the outside dimension of interconnecting device.
3. according to claim 2 for the device of nucleic acid rapid extraction, it is characterized in that, syringe comprises cylindrical shell, be placed in the tubulose Connection Block of barrel front end and be placed in cylindrical shell can push-and-pull push rod, the external diameter of described osculum pipe connecting equates with the Connection Block outside dimension of syringe, described interconnecting device is established intermediate throughholes, and intermediate throughholes size is mated with the Connection Block outside dimension of syringe.
According to described in claim 2 or 3 for the device of nucleic acid rapid extraction, it is characterized in that, described sample needle comprises needle stand and syringe needle, the external diameter of described osculum pipe connecting mates with the needle stand internal diameter of described sample needle.
According to described in claim 2 or 3 for the device of nucleic acid rapid extraction, it is characterized in that, interconnecting device one end is communicated with syringe, the other end is communicated with the osculum pipe connecting of adsorption film pipe, forms the drawing liquid mode of this device.
6. according to the device for nucleic acid rapid extraction described in claim 2 or 3 or 4, it is characterized in that, interconnecting device one end is communicated with syringe, and the other end is socketed in the opening end of adsorption film pipe, the osculum pipe connecting of adsorption film pipe is communicated with sample needle, forms the liquid mode that pushes away of this device.
7. according to claim 6 for the device of nucleic acid rapid extraction, it is characterized in that, described interconnecting device profile is round table-like, and the microcephaly of round table-like interconnecting device holds outside dimension to mate with the opening end of adsorption film pipe, and this opening end is held socket from the microcephaly of round table-like interconnecting device.
8. the method for a nucleic acid rapid extraction, right to use requires the device of 1 to 7 arbitrary described nucleic acid rapid extraction, after the steps such as sample cracking processing, adsorption film absorption sample amplifying nucleic acid, rinsing liquid rinsing adsorption film removal of impurities and elutriant wash-out nucleic acid, obtain pure nucleic acid solution.
9. method according to claim 8, is characterized in that, is " two take out one pushes away " method: " one takes out " is adsorbed on filter membrane the nucleic acid in lysate; " two take out " make rinsing liquid by filter membrane with clean impurity; " one pushes away " by the nucleic acid component wash-out being adsorbed on filter membrane, thereby has completed the extraction of biological specimen amplifying nucleic acid composition; Specifically comprise the following steps:
A) reagent indicates packing:
Each person-portion such as the lysate that may relate in nucleic acid extraction, RNA adsorption liquid, RNA enzyme inhibitors, 95% alcohol, 75% alcohol, rinsing liquid, elutriant is divided and is filled to 1.5EP pipe, and each pipe is indicated;
B) clinical sample is added in lysate pipe and (extract if relate to RNA, then add RNA adsorption liquid and RNA enzyme inhibitors), and mix;
C) one inhales: the drawing liquid form that uses nucleic acid-extracting apparatus, from sample cracking mixed solution, aspirate with the opening end of adsorption film pipe, the suction that utilizes syringe pump to produce, when nucleic acid component in sample is flowed through adsorption film because the specific adsorption of film is adsorbed on adsorption film, filtrate being has the waste liquid of the impurity such as cell debris and protein, and waste liquid enters syringe cavity and waste liquid only passes through this adsorption film once;
D) two inhale: the drawing liquid form that uses nucleic acid-extracting apparatus, from rinsing liquid pipe, aspirate with the opening end of adsorption film pipe, the suction that utilizes syringe pump to produce, clean adsorption film with rinsing liquid, cleaning remains in protein or other composition on adsorption film, and filtrate enters syringe cavity and this filtrate is only passed through adsorption film once;
E) one push away: the syringe of changing another cleaning, first suck elutriant, then use nucleic acid-extracting apparatus to push away liquid form (reverse by adsorption film pipe, make adsorption film front, opening end socket interconnecting device one end of adsorption film pipe, the osculum pipe connecting of adsorption film pipe connects the needle stand of sample needle), elutriant in syringe cavity is joined in adsorption film pipe, utilize pressure that syringe pushing produces by eluent stream through adsorption film, and the nucleic acid component wash-out being adsorbed on film is pushed to follow-up nucleic acid collection container or proofing unit, complete the leaching process of sample amplifying nucleic acid.
10. method according to claim 8, is characterized in that, is " three push away " method: " one pushes away " is adsorbed on filter membrane the nucleic acid in lysate; " two push away " makes rinsing liquid clean impurity by filter membrane; " three push away " by the nucleic acid component wash-out being adsorbed on filter membrane, thereby has completed the extraction of biological specimen amplifying nucleic acid composition; Specifically comprise the following steps:
A) reagent indicates packing:
Each person-portion such as the lysate that may relate in nucleic acid extraction, RNA adsorption liquid, RNA enzyme inhibitors, 95% alcohol, 75% alcohol, rinsing liquid, elutriant is divided and is filled to 1.5EP pipe, and each pipe is indicated to (using different letters and color differentiating);
B) clinical sample is added in lysate pipe and (extract if relate to RNA, then add RNA adsorption liquid and RNA enzyme inhibitors), and mix.
C) first suck sample cracking mixed solution with syringe, (adsorption film pipe is reverse for the liquid form that pushes away of use nucleic acid-extracting apparatus, do not establish sample needle), the pressure that utilizes syringe pushing to produce makes the cracking mixed solution filtering membrane of flowing through, when nucleic acid component in sample is flowed through adsorption film because the specific adsorption of film is adsorbed on adsorption film, filtrate being has the waste liquid of the impurity such as cell debris and protein, and waste liquid is discharged from and waste liquid only passes through adsorption film once;
D) suck again rinsing liquid with syringe, (adsorption film pipe is reverse for the liquid form that pushes away of use nucleic acid-extracting apparatus, do not establish sample needle), utilize pressure that syringe pushing produces that rinsing liquid is joined and in adsorption tube, cleans adsorption film, clear protein or other composition remaining on adsorption film, filtrate is discharged from and this filtrate is only passed through adsorption film once;
E) syringe of replacing another cleaning sucks elutriant, (adsorption film pipe is reverse for the liquid form that pushes away of use nucleic acid-extracting apparatus, add sample needle), elutriant in syringe cavity is joined in adsorption film pipe, utilize pressure that syringe pushing produces by eluent stream through adsorption film, and the nucleic acid component wash-out being adsorbed on film is pushed to follow-up nucleic acid collection container or proofing unit, complete the leaching process of sample amplifying nucleic acid.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004113042A (en) * | 2002-09-25 | 2004-04-15 | Fuji Photo Film Co Ltd | Nucleic acid-separating and purifying device |
| CN101684463A (en) * | 2008-09-28 | 2010-03-31 | 杭州优思达生物技术有限公司 | Method and kit for rapidly extracting nucleic acids from trace clinical samples |
| CN102409079A (en) * | 2010-08-26 | 2012-04-11 | 杭州优思达生物技术有限公司 | Novel infectious disease nucleic acid rapid detection kit and its detection method |
| CN203947102U (en) * | 2013-07-12 | 2014-11-19 | 中国人民解放军疾病预防控制所 | A kind of device for nucleic acid rapid extraction |
-
2014
- 2014-07-14 CN CN201410333236.7A patent/CN104059849B/en active Active
- 2014-07-14 CN CN201420386073.4U patent/CN203947102U/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004113042A (en) * | 2002-09-25 | 2004-04-15 | Fuji Photo Film Co Ltd | Nucleic acid-separating and purifying device |
| CN101684463A (en) * | 2008-09-28 | 2010-03-31 | 杭州优思达生物技术有限公司 | Method and kit for rapidly extracting nucleic acids from trace clinical samples |
| CN102409079A (en) * | 2010-08-26 | 2012-04-11 | 杭州优思达生物技术有限公司 | Novel infectious disease nucleic acid rapid detection kit and its detection method |
| CN203947102U (en) * | 2013-07-12 | 2014-11-19 | 中国人民解放军疾病预防控制所 | A kind of device for nucleic acid rapid extraction |
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|---|---|---|---|---|
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| WO2017067093A1 (en) * | 2015-10-21 | 2017-04-27 | 陈辉 | Nucleic acid extraction apparatus and extraction method |
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| CN203947102U (en) | 2014-11-19 |
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