CN101838609A - Sample adding device and application thereof - Google Patents
Sample adding device and application thereof Download PDFInfo
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- CN101838609A CN101838609A CN 201010124116 CN201010124116A CN101838609A CN 101838609 A CN101838609 A CN 101838609A CN 201010124116 CN201010124116 CN 201010124116 CN 201010124116 A CN201010124116 A CN 201010124116A CN 101838609 A CN101838609 A CN 101838609A
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Abstract
The invention provides a sample adding device which comprises a sample collecting area, a nucleic acid extracting area and a testing reagent sample adding area, integrates the functions of sample collection, nucleic acid extraction and testing reagent sample addition, and can be widely applied in automatic sample adding applications in sample analysis or testing in the fields of biology, immunology, medicine and the like, in particular to large-scale automatic analysis or screening. In addition, the invention further discloses a testing device comprising the sample adding device and a method for carrying out sample adding, analysis and testing.
Description
Technical field
The invention belongs to the bioassay technical field, particularly, the invention discloses sample adding device, it has integrated that sample compiles, the function of nucleic acid extraction and detection reagent application of sample, can be widely used in the sample analysis in fields such as biology, immunology, medical science or the automatic application of sample in the test and use, be particularly suited for large-scale the analysis automatically or examination.In addition, the invention also discloses test set that comprises sample adding device and the method for carrying out sample pipetting volume, analysis or test.
Background technology
In a large amount of clinical experiments in fields such as biology, immunology, medical science, medical jurisprudence and laboratory experiment, need to carry out based on polymerase chain reaction (PCR) and based on the analysis and the detection of immunity in conjunction with the biomacromolecule (as nucleic acid, protein etc.) of character.The application of sample process that in these tests, needs the transfer sample and/or test required reagent all essentially.Usually under less demanding condition, as when the small-scale clinical diagnosis, operator are direct control transfer pipet or use the lower automatic gear of degree of integration to carry out often, detects again if desired, needs directly pipette again initial sample.
For example, Chinese patent application 96111290.5 discloses a kind of automatic analysing apparatus that has dispenser, but this device does not have the nucleic acid extraction parts, can only be directly the sample handled (that is, the nucleic acid that has extracted) be tested, and need be carried out direct application of sample operation initial sample, do not compile sample and keep these secondary sample, if rollback detects again, must pipette initial sample again again, increase the contaminated chance of initial sample.
And for example, Chinese patent or patent application 200710042659.3 disclose the biological chips detection system of a kind of automatic sampling, application of sample, reaction, washing and detection, for automatic sampling, application of sample process, there are not the nucleic acid extraction parts equally yet, can only be directly to the sample handled (promptly, the nucleic acid that has extracted) test, and need carry out direct application of sample operation to initial sample, do not compile sample and keep these secondary sample, if rollback detects again, must pipette initial sample again again, increase the contaminated chance of initial sample.
For another example, Chinese patent 200720076920.7 discloses a kind of automatic nucleic acid extraction instrument, actual is exactly commercially available automatic magnetic bead extraction system, comprised " reagent discharges and the mixing pump system " that add reagent such as washing composition, because magnetic bead is small, in the process that holds and shift with magnetic force with bar magnet, having trace comes off and pollutes " reagent discharge and mixing pump system " and sample, pollute and be not easy to get rid of, this device do not combine by prompting and further buffering and application of sample parts yet in addition.
But for extensive (especially the low large scale test of positive rate, as blood screening), strict high application, the operation of prior art and device are also incompatible.For example, in famous U.S.'s Simpson uxoricide case, the final pollution that exists owing to the legal medical expert laboratory exactly causes legal medical expert's evidence of procuratorial organ unavailable in court; It is more and more higher that requirement detects in the country of large scale test projects such as blood screening, and still along with the popularization of welfares such as medical insurance, the monocyte sample of desired input detects cost and but is required to reduce.The inventor through a long-term line investigation find, when low and testing sample number is many when positive rate, each single sample and uneconomical that detects separately, efficient is not high; Do not keep and compile sample,, must return application of sample initial sample again once more, increase the number of times of sampling initial sample, increased the chance of polluting for when detected result is unusual; Although the magnetic bead extraction system is simple and convenient, wherein the easiest magnetic bead that causes pollutes in the process that shifts magnetic bead with bar magnet, needs or reusable sample of potential demand and reagent the time, this pollution will be difficult to get rid of if pollute; If adopt special parts to prevent to pollute, will improve the cost of final loading device greatly, more be difficult on extensive project, apply.Therefore, the inventor is through long-term and arduous research, research has obtained a kind of layout of sample adding device, it has integrated that sample compiles, the function of nucleic acid extraction and detection reagent application of sample, not only disposablely finishes above process, and has avoided pollution by layout as far as possible, need not to use special parts, and have a large amount of parts can be general, and made things convenient for parts buying, device to make and device is safeguarded, can effectively reduce cost.In addition, the inventor has also obtained to comprise the test set of sample adding device and the method for carrying out sample pipetting volume, analysis or test.
Summary of the invention
It is high, uneconomical and potential shortcoming such as more pollution (especially be difficult to the pollution of getting rid of and the initial sample pollution that is difficult to retrieve etc.) may take place that the technical problem to be solved in the present invention is to overcome prior art efficient in extensive biomacromolecule test (the especially low large scale test of positive rate), provides and integrated that sample compiles, the sample adding device of nucleic acid extraction and detection reagent application of sample function.The obtained effect of this device not only can overcome defective of the prior art as much as possible, and can not use the particular component manufacturing, wherein a large amount of in addition parts can be general, made things convenient for parts buying, device to make and the device maintenance, thereby made manufacturing cost and use cost cheaper.
Particularly, aspect first, the invention provides sample adding device, it comprises test tube, sample adding device shell, dispenser, stirring rod and bar magnet, it is characterized in that, comprises sample pooling zone, nucleic acid extraction district and test agent sample application zone in the described sample adding device shell, wherein
The sample pooling zone comprises dispenser, sample liquid storage tank (11), disposable transfer pipet storage tank (12), transfer pipet disposal receptacle (13), compiles the storage tank (15) of sample liquid storage tank (14), reagent storage groove (16) and removable tote, and cuts off by the dividing plate (10) that has door (101) between sample pooling zone and the nucleic acid extraction district;
The nucleic acid extraction district comprises the storage tank (25) of storage tank (21), bar magnet cover storage tank (22), magnetic bead storage tank (24), washing lotion storage tank (23) and the second removable tote of stirring rod, bar magnet, the first removable tote;
The test agent sample application zone comprises dispenser, storage tank (41), disposable transfer pipet storage tank (42), transfer pipet disposal receptacle (43), reagent storage groove (45) and application of sample mixed solution storage tank (44);
And, the door (101) that the storage tank (21) of the first removable tote in the storage tank of the removable tote in the sample pooling zone (15) and the nucleic acid extraction district has across dividing plate (10) and adjacent, and the storage tank of the removable tote in the sample pooling zone (15) can move the test-tube stent (151) of its loading on the storage tank (21) in the first removable tote that is loaded into the nucleic acid extraction district, and the storage tank (21) of the first removable tote in the nucleic acid extraction district also can move the test-tube stent (151) of its loading and be loaded on the storage tank (15) of the removable tote in the sample pooling zone; The storage tank (25) of the second removable tote in the nucleic acid extraction district loads elutriant test-tube stent (251), and described elutriant test-tube stent (251) can be moved on the storage tank (41) in the test agent sample application zone.
" first " herein, " second " be just to the difference of distinguishing entity, and do not represent the difference of entity structure.
" sample " herein is meant the liquid that employed band detects in the analysis, test of biology, immunology, medical field, as blood, saliva, tissue juice, equal slurries, preferred this sample can pass through pre-treatment, condenses preventing as adding heparin in the blood.
" storage tank " herein, as do not have opposite indication is not limited to only refer to container (as loading the groove shape container of test-tube stent), also refers to upholder, as the plate that supports test-tube stent, plane etc.Herein " storage tank of removable tote " refers to storage tank, and it has, and the tote that can make its loading is directed thereon to be moved or the directed parts that shift out it, thereby can mobile tote.Wherein said parts include but not limited to slide rail, ball, push rod etc.Disposable transfer pipet storage tank is preferably directly deposited one or more sockets disposable transfer pipet up; Bar magnet cover storage tank is preferably directly deposited one or more sockets bar magnet cover up; Other storage tanks can directly be separated out the space in order to load one or more test tubes, but load test-tube stent on the preferred storage tank, load one or more test tube mouths test tube up in described test-tube stent, described test tube has added sample, reagent, magnetic bead or magnetic bead suspension, washing lotion or elutriant respectively according to the qualification of the deposit of storage tank or test-tube stent; Described test tube also can be empty, thereby can be added into several samples liquid so that accumulate and compile sample liquid, perhaps is added into and compiles sample liquid, reagent, magnetic bead and/or their mixed solution etc.In the sample adding device of first aspect present invention, the storage tank of removable tote can be identical on the block construction, just optional travel direction difference of settling, so these parts can be general mutually; Other storage tanks can be identical or can be to have an identical repeating unit on the block construction, so also can be general mutually or assembly unit, disassemble.This all greatly facilitates makes and maintenance.The storage tank (21) of the first removable tote in the preferred nucleic acid extraction district has heating unit in addition, thereby can the sample liquid of compiling that add the nucleic acid lysate be heated, and quickens the cracking of sample.
The transfer pipet disposal receptacle is to deposit the container of the disposable transfer pipet of throwing aside.The transfer pipet disposal receptacle can have opening at an upper portion thereof, and preferably less opening as can vertically falling into the opening of transfer pipet disposal receptacle inside for disposable transfer pipet, can be avoided disposable transfer pipet to rebound out internally so as far as possible and pollute other zones.Disposable transfer pipet is meant disposable transfer pipet in sample adding device of the present invention, but this transfer pipet is not got rid of to reclaim separately and cleaned the back and play a role again in follow-up use.Disposable transfer pipet can be made with suitable material, is made of plastics usually, as those skilled in the familiar " Tip head " or " rifle head " etc.
In the sample pooling zone, usually a sample in the sample liquid storage tank (11) is only carried out primary sample, sample is added in the test tube that compiles in the sample liquid storage tank (14), avoid resampling from sample liquid storage tank (11) later on as far as possible, thereby reduce the contaminated chance of initial sample; And (as be lower than 1% for positive rate is low, or be lower than thousandth, ten thousand/first-class) large scale test, different samples can be added in the same test tube that compiles in the sample liquid storage tank (14), as with 5,10,15,20 or a plurality of samples of other quantity add in the same test tube that compiles in the sample liquid storage tank (14), the sample liquid of compiling in the test tube in the sample liquid storage tank (14) of compiling is handled and tested, can avoid a large amount of samples to repeat to draw a large amount of negative findingses, effectively reduce and handle and test quantity, therefore preferably the test tube number that can deposit of sample liquid storage tank (11) will be far away more than compiling the stored test tube number of sample liquid storage tank (14).For example, the test tube number that sample liquid storage tank (11) can be deposited is 2500, and the sample number that can deposit can be less than 2500, and as 2304, compiling the stored test tube number of sample liquid storage tank (14) is 250, can compile with 1: 10 ratio.
Reagent herein, magnetic bead, washing lotion, elutriant be those skilled in the art according to the purpose of nucleic acid extraction, test can understand, and can buy by market channel, also can be with reference to Chinese patent application 96197315.3,00811856.6,200610030229.5 etc.For example, the reagent of depositing in the reagent storage tank (16) in the sample pooling zone comprises the nucleic acid lysate and in conjunction with liquid.The nucleic acid lysate normally contains the solution of guanidine class (being preferably guanidine thiocyanate); Normally contain the chaotropic salt ion solution of high density in conjunction with liquid, make the chaotropic salt ionic concentration of mixed solution of biomacromolecule and magnetic bead be between the 0.8-4.5M, thereby make biomacromolecule be adsorbed on the magnetic bead." magnetic bead " used herein and " magnetic-particle ", " magnetic colloid " are general, refer to particle diameter and be adsorbing biomacromolecule and having the granule of magnetic of 0.1-100 micron (being preferably the 1-10 micron), for example the granule of polymkeric substance parcel magnet.The washing lotion of magnetic bead normally contains the aqueous solution of salt (as acetate) and alcohol (as ethanol), can prevent adsorptive on the magnetic-particle when cleaning by wash-out.The elutriant of the adsorptive aqueous solution of ionic strength normally on the magnetic bead.The reagent of depositing in the reagent storage tank (45) in the test agent sample application zone is test agent.In a specific embodiment of the present invention, test agent is the PCR test agent, comprises amplified reaction damping fluid, dNTP mixing solutions, dna probe solution, primed DNA solution and thermostability archaeal dna polymerase solution.
Dispenser, stirring rod and bar magnet are that those skilled in the art use always, can buy by market channel.These parts can carry out X, Y, Z three directions move, thereby can move to suitable position.Dispenser, stirring rod and bar magnet can be installed in respectively on the mechanical arm, by mechanical arm be controlled at X, Y, Z three directions move.But preferably in described sample adding device, because storage tanks etc. are fixed normally, therefore can adopt the more cheap form that adds Z axle motor as the guide rail of Chinese patent application 200710042659.3, be that wherein said dispenser, stirring rod and bar magnet are installed in respectively on the guide rail, can carry out X, Y respectively, Z three directions move.In addition, bar magnet can be an electromagnet type, is in the state that does not have magnetic when no power, is in magnetic state when energising; Described as Chinese patent 200720076920.7, bar magnet also can be removable, promptly is surrounded by the not pipe box of shielding magnetic in the magnet periphery, is in the state that does not have magnetic when magnet is extracted out, is in magnetic state when magnet inserts.
Dispenser, stirring rod and bar magnet be form side by side preferably, it is preferably hyperchannel dispenser of dispenser, many stirring rod are arranged into a row and move together, many bar magnets are arranged into a row and move together, when test tube, disposable transfer pipet or the magnetic rod sleeve in the storage tank also is corresponding form and is arranged side by side like this, can once move and simultaneously these test tubes, disposable transfer pipet or magnetic rod sleeve are operated.Maybe can go deep into beginning operation near the test tube bottom because the liquid level that stirring rod and bar magnet need be operated usually is a fixed, so they need not supporting liquid level detector usually; And dispenser, especially when drawing sample, sample liquid level in the test tube often has error, therefore the dispenser in the preferred described sample pooling zone has air pressure type liquid level detector, avoid being subjected to the influence of sample liquid level and too much deeply cause disposable transfer pipet outer wall to be infected with too much sample, dispenser in the described in addition sample adding device can have air pressure type liquid level detector, makes things convenient for component maintenance.In addition, also preferred dispenser can have baffle plate, and stirring rod has baffle plate, can avoid shifting out the liquid that sticks on the outer wall behind the liquid and drip.Baffle plate keeps off in the bottom in the dispenser moving process, and when dispenser will move down with absorption or inject liquid, baffle plate was removed not hinder dispenser and moved down.The mode that baffle plate is removed can be horizontal translation or rotate to a side.More preferably the stirring rod in the nucleic acid extraction district has heating unit, thereby can the sample liquid of compiling that add the nucleic acid lysate be heated when stirring, and quickens the cracking of sample.
Herein " door " is meant the parts that can open with closed channel, but preferred herein door is the door of translation, but conserve space thus.The storage tank of removable tote is adjacent with another storage tank that can accept tote, therefore when opening, by tote is advanced on another storage tank that can accept tote on the storage tank of removable tote such as the active role of ball, push rod; And when door when closing, can play isolation action and reduce pollution.
The storage tank (25) of the second removable tote in nucleic acid extraction district can be adjacent with the storage tank (41) in the test agent sample application zone, therefore elutriant test-tube stent (251) directly can be moved on the storage tank (41) in the test agent sample application zone, the storage tank (25) of the second removable tote in optional nucleic acid extraction district can and the test agent sample application zone in storage tank (41) between cut off by the added partition that has door, by the sealing process of dividing plate and door, further reduce the chance of polluting.
If but we find and can isolate static for some time, extremely is helped further to reduce to pollute, be that tiny magnetic bead is floated to the test agent sample application zone thus, therefore in the sample adding device of first aspect present invention, also comprise buffer zone in the preferred described sample adding device shell, described buffer zone comprises the storage tank (31) of removable tote:
Cut off the door (201) that the storage tank (31) of the removable tote in the storage tank (25) of the second removable tote in the nucleic acid extraction district and the buffer zone has across first added partition (20) and adjacent between described buffer zone and the nucleic acid extraction district by first added partition (20) that has door (201); Cut off the door (301) that the storage tank (41) in the storage tank of the removable tote in the buffer zone (31) and the test agent sample application zone has across second added partition (30) and adjacent between described buffer zone and the test agent sample application zone by second added partition (30) that has door (301);
And, the storage tank (25) of the second removable tote in the nucleic acid extraction district can move the elutriant test-tube stent (251) of its loading and be loaded on the storage tank (31) of the removable tote in the buffer zone, and the storage tank of the removable tote in the buffer zone (15) can move the elutriant test-tube stent (251) of its loading on the storage tank (41) that is loaded in the test agent sample application zone.
" added partition " herein and " dividing plate " be just to distinguishing the different of dividing plate entity, and do not represent the difference of diaphragm structure, " auxiliary washing lotion storage tank " and " washing lotion storage tank " too.By dividing plate, independently sample pooling zone, nucleic acid extraction district, buffer zone, test agent sample application zone and buffer zone have been separated out separately in described sample adding device inside.For further antipollution chance, preferred sample pooling zone, nucleic acid extraction district, buffer zone and/or test agent sample application zone comprise the gas barrier that has filter screen.Wherein said filter screen aperture is less than 1 μ m, is preferably 0.2-0.5 μ m, most preferably is 0.3 μ m.Can effectively prevent from like this to pollute.In addition, buffer zone also can comprise the gas barrier that has filter screen.
The layout of some details also can obtain the lifting of sizable preventing polluting effect under the condition that does not improve manufacturing cost substantially.For example, in the sample pooling zone, sample liquid storage tank (11) lays respectively at the not homonymy that compiles sample liquid storage tank (14) with these two parts of storage tank (15) of removable tote.Like this, draw compile sample liquid and dispenser needn't pass through sample liquid storage tank (11), can effectively prevent from moving process, to compile sample liquid and drip sample in the contaminated samples liquid storage tank (11).Equally, also preferred reagent storage tank (16) is positioned at the homonymy that compiles sample liquid storage tank (14) with these two parts of storage tank (15) of removable tote, also can avoid reagent to drip sample in the contaminated samples liquid storage tank (11) in the process of pipetting.
And for example, the storage tank (25) of the storage tank (21) of the first removable tote in the nucleic acid extraction district, bar magnet cover storage tank (22), magnetic bead storage tank (24), washing lotion storage tank (23) and the second removable tote is arranged in order, especially linear arrangement.So make things convenient for the setting and the economy of guide rail, the feasible on the other hand operation at each storage tank does not move through on other storage tanks as far as possible, pollutes thereby reduce.Also preferred in addition bar magnet has many or many rows, can be respectively at the operation of some storage tank.Preferred nucleic acid extracts to distinguish and also comprises the first auxiliary washing lotion storage tank (26) and the second auxiliary washing lotion storage tank (27), in order to the washing lotion of adding different concns, thereby constitutes the cleaning gradient with washing lotion storage tank (23).Wherein, more preferably the storage tank (25) of the storage tank of the first removable tote (21), bar magnet cover storage tank (22), magnetic bead storage tank (24), washing lotion storage tank (23) first auxiliary washing lotion storage tanks (26), the second auxiliary washing lotion storage tank (27) and the second removable tote is arranged in order, especially linear arrangement.Test tube in preferred each washing lotion storage tank and the test tube in the elutriant test-tube stent (251) are added with washing lotion and elutriant respectively, complicated reagent needn't be set like this pipette device, and when polluting, directly change the test tube that is added with washing lotion and elutriant respectively and get final product.
Therefore the sample adding device of first aspect present invention can be controlled automatically, and preferred described sample adding device comprises and can control dispenser, stirring rod and bar magnet operation, can control the automatic control component that storage tank that door that each dividing plate has opened and closed and can control each removable tote moves tote.Wherein, described operation comprises the volume that moves (or stirring), dispenser absorption and emit and draw and emit of X, Y, Z direction, controls having or not of bar magnet magnetic force etc.Automatic control component can be a micro-chip, but preferably can adopt commercially available computer.In addition, the sample pooling zone preferably also comprises and has the bar code recognition device, the barcode of identification sample tube, the control automatically of being more convenient for.
Aspect second, the invention provides sample adding device biased sample and compositions and methods with the present invention first aspect, it is characterized in that described method comprises,
(a1) in the sample pooling zone, dispenser moves and plugs the disposable transfer pipet of depositing in the disposable transfer pipet storage tank (12) successively, move and suck the sample liquid of depositing in the sample liquid storage tank (11), move and sample liquid is injected the test tube that compiles sample liquid storage tank (14), move and disposable transfer pipet is thrown aside in transfer pipet disposal receptacle (13);
(a2) then, in the sample pooling zone, dispenser moves and plugs the disposable transfer pipet of depositing in the disposable transfer pipet storage tank (12) successively, move and suck the nucleic acid lysate in the reagent storage groove (16), move and the nucleic acid lysate is injected the test tube of the test-tube stent (151) that the storage tank (15) of removable tote loads, move and disposable transfer pipet is thrown aside in transfer pipet disposal receptacle (13); In the sample pooling zone, dispenser moves and plugs the disposable transfer pipet of depositing in the disposable transfer pipet storage tank (12) successively, move and suck and compile the sample liquid of depositing in the sample liquid storage tank (14) of compiling, move and will compile the test tube that sample liquid is injected the test-tube stent (151) that the storage tank of removable tote (15) loads, move and disposable transfer pipet is thrown aside in transfer pipet disposal receptacle (13);
(a3) then, open the door (101) of the dividing plate (10) between sample pooling zone and the nucleic acid extraction district, the storage tank of the removable tote in the sample pooling zone (15) moves the test-tube stent (151) of its loading at the door (101) that the dividing plate (10) between sample pooling zone and the nucleic acid extraction district was gone up and closed to storage tank (21) in the first removable tote that is loaded into the nucleic acid extraction district, in the nucleic acid extraction district, stirring rod is moved and stirs the liquid in the test tube of described test-tube stent (151), retract stirring rod, open the door (101) of the dividing plate (10) between sample pooling zone and the nucleic acid extraction district, the storage tank (21) in the first removable tote in nucleic acid extraction district moves the door (101) that the dividing plate (10) between sample pooling zone and the nucleic acid extraction district was gone up and closed to the storage tank (15) that is loaded into the removable tote in the sample pooling zone with the test-tube stent (151) of its loading;
(a4) then, in the sample pooling zone, dispenser moves and plugs the disposable transfer pipet of depositing in the disposable transfer pipet storage tank (12) successively, move and suck in the reagent storage groove (16) in conjunction with liquid, move and will inject the test tube of the test-tube stent (151) that the storage tank (15) of removable tote loads, move and disposable transfer pipet is thrown aside in transfer pipet disposal receptacle (13) in conjunction with liquid;
(a5) then, open the door (101) of the dividing plate (10) between sample pooling zone and the nucleic acid extraction district, the storage tank of the removable tote in the sample pooling zone (15) moves the test-tube stent (151) of its loading at the door (101) that the dividing plate (10) between sample pooling zone and the nucleic acid extraction district was gone up and closed to storage tank (21) in the first removable tote that is loaded into the nucleic acid extraction district;
(b1) then, in the nucleic acid extraction district, bar magnet moves and puts the bar magnet cover of depositing in the bar magnet cover storage tank (22) successively, under magnetic state, move and absorption magnetic bead storage tank (24) in the magnetic bead deposited, under magnetic state, move and insert in the test tube of the test-tube stent (151) that the storage tank (21) of the first removable tote loads, magnetic disappears so that adsorbed magnetic bead comes off, and moves and the bar magnet cover is retracted in the bar magnet cover storage tank (22);
(b2) then, in the nucleic acid extraction district, stirring rod is moved and stirs the liquid in the test tube of the test-tube stent (151) that the storage tank (21) of the first removable tote loads, retract stirring rod;
(b3) then, in the nucleic acid extraction district, bar magnet moves and puts the bar magnet cover of depositing in the bar magnet cover storage tank (22) successively, under magnetic state, move and adsorb the magnetic bead in the test tube of the test-tube stent (151) that the storage tank (21) of the first removable tote loads, under magnetic state, move and insert in the washing lotion in the washing lotion storage tank (23), magnetic disappears so that adsorbed magnetic bead comes off, and moves and the bar magnet cover is retracted in the bar magnet cover storage tank (22);
(b4) then, in the nucleic acid extraction district, bar magnet under magnetic state, move successively and absorption washing lotion storage tank (23) in washing lotion in magnetic bead, move under magnetic state and insert in the test tube in the elutriant test-tube stent (251) that the storage tank (25) of the second removable tote loads, magnetic disappears so that adsorbed magnetic bead comes off;
(b5) in the nucleic acid extraction district, operate in or afterwards, in the test agent sample application zone, dispenser moves and plugs the disposable transfer pipet of depositing in the disposable transfer pipet storage tank (42) successively, move and suck the detection reagent in the reagent storage groove (45), move and detection reagent is injected the test tube of application of sample mixed solution storage tank (44), move and disposable transfer pipet is thrown aside in transfer pipet disposal receptacle (43);
(c) then, the elutriant test-tube stent (251) that the storage tank (25) of the second removable tote in the nucleic acid extraction district is loaded is moved on the storage tank (41) in the test agent sample application zone;
(d) then, in the test agent sample application zone, dispenser moves and plugs the disposable transfer pipet of depositing in the disposable transfer pipet storage tank (42) successively, move and suck the liquid in the test tube in the elutriant test-tube stent (251) that storage tank (41) loads, move and described liquid is injected the test tube of application of sample mixed solution storage tank (44), move and disposable transfer pipet is thrown aside in throwing aside in the container (43).
Preferably in step (c), open the door (201) of first added partition (20) between buffer zone and the nucleic acid extraction district, the storage tank (25) of the second removable tote in the nucleic acid extraction district moves the elutriant test-tube stent (251) of its loading and is loaded on the storage tank (31) of the removable tote in the buffer zone, close the door (201) of first added partition (20) between buffer zone and the nucleic acid extraction district, open the door (301) of second added partition (30) between buffer zone and the test agent sample application zone after leaving standstill, the storage tank of the removable tote in the buffer zone (31) moves the elutriant test-tube stent (251) of its loading on the storage tank (41) that is loaded in the test agent sample application zone, closes the door (301) of second added partition (30) between buffer zone and the test agent sample application zone.Preferred in addition in the step of lysate sample, promptly in (a3) step, stirring rod heats described liquid when stirring liquid in the test tube of described test-tube stent (151).
In the method for a second aspect of the present invention, can choose each step of repetition wantonly, for example in (a1),, can repeat (a1) step for mobile several samples.In addition, before the method for a second aspect of the present invention is carried out, the Men Juncheng closing condition, sample, reagent, washing lotion, magnetic bead, elutriant all need to add in the test tube on the corresponding storage tank, wherein said test-tube stent (151) is loaded on the storage tank (15) of the removable tote in the sample pooling zone, and described elutriant test-tube stent (251) is loaded on the storage tank (25) of the second removable tote in the nucleic acid extraction district.These all are understandable to those skilled in the art.
Aspect the 3rd, the invention provides the nucleic acid test set, it is characterized in that described device comprises described sample adding device in first aspect of the present invention and nucleic acid tester.Described nucleic acid test set can comprise the transhipment parts, and the application of sample mixed solution that final application of sample can be finished is transported in the described nucleic acid tester, and preferred described transhipment parts are dispensers.But the application of sample mixed solution storage tank (44) in the described sample adding device in first aspect of the present invention in the preferred described nucleic acid test set just is installed on the described nucleic acid tester, can save the transhipment parts like this.
In the nucleic acid test set of third aspect of the present invention, preferred described nucleic acid tester is the PCR instrument, more preferably the application of sample mixed solution storage tank (44) in the described sample adding device in first aspect of the present invention just is installed on the described PCR instrument, and more preferably described PCR instrument is also controlled automatically.
Aspect the 4th, the invention provides method with the nucleic acid test set analytic sample of third aspect of the present invention, it is characterized in that described method comprises the described biased sample and the compositions and methods of carrying out second aspect of the present invention, detect nucleic acid and analysis with the nucleic acid tester then.
Below will by the drawings and specific embodiments to sample adding device of the present invention, add quadrat method and comprise that the nucleic acid test set of this sample adding device carries out exemplary description.In addition, the open source literature of quoting is herein all included this paper reference in, just looks like that they repeat to tell the same in this article.
Description of drawings
The exemplary whole lower floor schematic layout pattern of Fig. 1 sample adding device of the present invention.
Exemplary lower floor's schematic layout pattern of sample pooling zone in Fig. 2 sample adding device of the present invention.
Fig. 3 sample adding device amplifying nucleic acid of the present invention is extracted exemplary lower floor's schematic layout pattern in district.
Exemplary lower floor's schematic layout pattern of test agent sample application zone in Fig. 4 sample adding device of the present invention.
Embodiment
Following according to the layout shown in Fig. 1-4 to sample adding device of the present invention, add quadrat method and comprise that the nucleic acid test set of this sample adding device carries out exemplary description.
Embodiment 1 sample adding device
Exemplary sample adding device comprises test tube, sample adding device shell, dispenser, stirring rod and bar magnet, it is characterized in that, comprises sample pooling zone, nucleic acid extraction district, buffer zone and test agent sample application zone in the described sample adding device shell, wherein,
The sample pooling zone comprises the dispenser that is positioned on the guide rail of upper strata, and include: comprise a plurality of sample liquid storage tanks 11 of depositing the test tube 111 of sample liquid in lower floor, the disposable transfer pipet storage tank 12 that comprises a plurality of disposable transfer pipets 121, transfer pipet disposal receptacle 13, what comprise a plurality of test tubes 141 compiles sample liquid storage tank 14, comprise and a plurality ofly deposit the nucleic acid lysate at least and in conjunction with the reagent storage groove 16 of the test tube 161 of liquid be mounted with the storage tank 15 of the removable tote of test-tube stent 151, a plurality of test tubes 152 are housed on the described test-tube stent 151, and cut off by the dividing plate 10 that has door 101 between sample pooling zone and the nucleic acid extraction district;
The nucleic acid extraction district comprises stirring rod and the bar magnet that is positioned on the guide rail of upper strata, and include successively: the storage tank 21 of the first removable tote in lower floor, the bar magnet cover storage tank 22 that comprises a plurality of bar magnet covers, comprise a plurality of storage tanks 24 of depositing the test tube of magnetic bead suspension, comprise a plurality of washing lotion storage tanks 23 of depositing the test tube of the first concentration washing lotion, comprise a plurality of first auxiliary washing lotion storage tanks 26 of depositing the test tube of the second concentration washing lotion, comprise a plurality of second auxiliary washing lotion storage tanks 27 of depositing the test tube of the 3rd concentration washing lotion, with the storage tank 25 of the second removable tote that is mounted with elutriant test-tube stent 251, a plurality of test tubes 252 of depositing elutriant are housed on the described elutriant test-tube stent 251;
Buffer zone comprises the storage tank 31 of removable tote; And, cut off the door 201 that the storage tank 31 of the removable tote in the storage tank 25 of the second removable tote in the nucleic acid extraction district and the buffer zone has across first added partition 20 and adjacent between described buffer zone and the nucleic acid extraction district by first added partition 20 that has door 201; Cut off the door 301 that the storage tank 41 in the storage tank 31 of the removable tote in the buffer zone and the test agent sample application zone has across second added partition 30 and adjacent between described buffer zone and the test agent sample application zone by second added partition 30 that has door 301;
The test agent sample application zone comprises the dispenser that is positioned on the guide rail of upper strata, and include in lower floor: storage tank 41, comprise a plurality of disposable transfer pipets 421 disposable transfer pipet storage tank 42, transfer pipet disposal receptacle 43, comprise a plurality of PCR of depositing reagent test tube 451 reagent storage groove 45 and comprise the application of sample mixed solution storage tank 44 of test-tube stent 441, a plurality of test tubes 442 are housed on the described test-tube stent 441;
And, the door 101 that the storage tank 21 of the first removable tote in the storage tank 15 of the removable tote in the sample pooling zone and the nucleic acid extraction district has across dividing plate 10 and adjacent, and the storage tank 15 of the removable tote in the sample pooling zone can move the test-tube stent 151 of its loading on the storage tank 21 in the first removable tote that is loaded into the nucleic acid extraction district, if load, the storage tank 21 of the first removable tote in the nucleic acid extraction district also can move the test-tube stent 151 of its loading on the storage tank 15 that is loaded into the removable tote in the sample pooling zone; The storage tank 25 of the second removable tote in the nucleic acid extraction district can move the elutriant test-tube stent 251 of its loading on the storage tank 31 that is loaded into the removable tote in the buffer zone, and the storage tank 15 of the removable tote in the buffer zone can move the elutriant test-tube stent 251 of its loading on the storage tank 41 that is loaded in the test agent sample application zone.
Embodiment 2 adds quadrat method
According to embodiment 1 described sample adding device, before carrying out application of sample, the Men Juncheng closing condition, sample, reagent, washing lotion, magnetic bead, elutriant all add respectively in the test tube on the corresponding storage tank, wherein said test-tube stent 151 is loaded on the storage tank 15 of the removable tote in the sample pooling zone, and described elutriant test-tube stent 251 is loaded on the storage tank 25 of the second removable tote in the nucleic acid extraction district.
May further comprise the steps when carrying out application of sample:
(a1) in the sample pooling zone, dispenser moves and plugs the disposable transfer pipet of depositing in the disposable transfer pipet storage tank 12 successively, move and suck the sample liquid of depositing in the sample liquid storage tank 11, move and sample liquid is injected the test tube that compiles sample liquid storage tank 14, move and disposable transfer pipet is thrown aside in transfer pipet disposal receptacle 13;
(a2) then, in the sample pooling zone, dispenser moves and plugs the disposable transfer pipet of depositing in the disposable transfer pipet storage tank (12) successively, move and suck the nucleic acid lysate in the reagent storage groove 16, move and the nucleic acid lysate is injected the test tube of the test-tube stent 151 that the storage tank 15 of removable tote loads, move and disposable transfer pipet is thrown aside in transfer pipet disposal receptacle 13; In the sample pooling zone, dispenser moves and plugs the disposable transfer pipet of depositing in the disposable transfer pipet storage tank 12 successively, move and suck and compile the sample liquid of depositing in the sample liquid storage tank 14 of compiling, move and will compile the test tube that sample liquid is injected the test-tube stent 151 that the storage tank 15 of removable tote loads, move and disposable transfer pipet is thrown aside in transfer pipet disposal receptacle 13;
(a3) then, open the door 101 of the dividing plate 10 between sample pooling zone and the nucleic acid extraction district, the storage tank 15 of the removable tote in the sample pooling zone moves the test-tube stent 151 of its loading on the storage tank 21 in the first removable tote that is loaded into the nucleic acid extraction district and closes the door 101 of the dividing plate 10 between sample pooling zone and the nucleic acid extraction district, in the nucleic acid extraction district, stirring rod is moved and stirs the liquid in the test tube of described test-tube stent 151, retract stirring rod, open the door 101 of dividing plate 10 between sample pooling zone and the nucleic acid extraction district, the storage tank 21 in the first removable tote in nucleic acid extraction district moves the test-tube stent 151 of its loading on the storage tank 15 that is loaded into the removable tote in the sample pooling zone and closes the door 101 of the dividing plate 10 between sample pooling zone and the nucleic acid extraction district;
(a4) then, in the sample pooling zone, dispenser moves and plugs the disposable transfer pipet of depositing in the disposable transfer pipet storage tank 12 successively, move and suck in the reagent storage groove 16 in conjunction with liquid, move and will inject the test tube of the test-tube stent 151 that the storage tank 15 of removable tote loads, move and disposable transfer pipet is thrown aside in transfer pipet disposal receptacle 13 in conjunction with liquid;
(a5) then, open the door 101 of dividing plate 10 between sample pooling zone and the nucleic acid extraction district, the storage tank 15 of the removable tote in the sample pooling zone moves the test-tube stent 151 of its loading on the storage tank 21 in the first removable tote that is loaded into the nucleic acid extraction district and closes the door 101 of the dividing plate 10 between sample pooling zone and the nucleic acid extraction district;
(b1) then, in the nucleic acid extraction district, bar magnet moves and puts the bar magnet cover of depositing in the bar magnet cover storage tank 22 successively, under magnetic state, move and absorption magnetic bead storage tank 24 in the magnetic bead deposited, under magnetic state, move and insert in the test tube of the test-tube stent 151 that the storage tank 21 of the first removable tote loads, magnetic disappears so that adsorbed magnetic bead comes off, and moves and the bar magnet cover is retracted in the bar magnet cover storage tank 22;
(b2) then, in the nucleic acid extraction district, stirring rod is moved and stirs the liquid in the test tube of the test-tube stent 151 that the storage tank 21 of the first removable tote loads, retract stirring rod;
(b3) then, in the nucleic acid extraction district, bar magnet moves and puts the bar magnet cover of depositing in the bar magnet cover storage tank 22 successively, under magnetic state, move and adsorb the magnetic bead in the test tube of the test-tube stent 151 that the storage tank 21 of the first removable tote loads, under magnetic state, move and insert in the washing lotion in the washing lotion storage tank 23, magnetic disappears so that adsorbed magnetic bead comes off, and moves and the bar magnet cover is retracted in the bar magnet cover storage tank 22;
(b4) then, in the nucleic acid extraction district, bar magnet under magnetic state, move successively and absorption washing lotion storage tank 23 in washing lotion in magnetic bead, move under magnetic state and insert in the test tube in the elutriant test-tube stent 251 that the storage tank 25 of the second removable tote loads, magnetic disappears so that adsorbed magnetic bead comes off;
(b5) in the nucleic acid extraction district, operate in or afterwards, in the test agent sample application zone, dispenser moves and plugs the disposable transfer pipet of depositing in the disposable transfer pipet storage tank 42 successively, move and suck the detection reagent in the reagent storage groove 45, move and detection reagent is injected the test tube of application of sample mixed solution storage tank 44, move and disposable transfer pipet is thrown aside in transfer pipet disposal receptacle 43;
(c) then, open the door 201 of first added partition 20 between buffer zone and the nucleic acid extraction district, the storage tank 25 of the second removable tote in the nucleic acid extraction district moves the elutriant test-tube stent 251 of its loading on the storage tank 31 that is loaded into the removable tote in the buffer zone, close the door 201 of first added partition 20 between buffer zone and the nucleic acid extraction district, open the door 301 of second added partition 30 between buffer zone and the test agent sample application zone after leaving standstill, the storage tank 31 of the removable tote in the buffer zone moves the elutriant test-tube stent 251 of its loading on the storage tank 41 that is loaded in the test agent sample application zone, closes the door 301 of second added partition 30 between buffer zone and the test agent sample application zone;
(d) then, in the test agent sample application zone, dispenser moves and plugs the disposable transfer pipet of depositing in the disposable transfer pipet storage tank 42 successively, move and suck the liquid in the test tube in the elutriant test-tube stent 251 that storage tank 41 loads, move and described liquid is injected the test tube of application of sample mixed solution storage tank 44, move and disposable transfer pipet is thrown aside in transfer pipet disposal receptacle 43.
Embodiment 3 nucleic acid test sets
Exemplary nucleic acid test set comprises embodiment 1 described sample adding device and PCR instrument, and the application of sample mixed solution storage tank 44 in the described sample adding device just is installed on the PCR instrument.
Embodiment 4 PCR test results
, the result
The sample of known HBV DNA concentration is diluted to 10
5IU/ml, with the sample of this concentration and negative sample on the sample disk with the bigger arrangement mode of mutual pollution probability---after being staggered, with embodiment 3 described nucleic acid test sets these samples are carried out nucleic acid extraction and pcr amplification detection, the result conforms to fully with the yin and yang attribute result of expection.Wherein the one group of result who selects at random is shown in following table 1 and 2, and the result shows that apparatus of the present invention can effectively be carried out the sample nucleic acid extraction, prevent crossed contamination between sample.This shows that nucleic acid test set of the present invention can effectively prevent to pollute, and can apply in the demanding biomacromolecule test.
Table 1: the arrangement mode of sample in sample disk
| ??1 | ??2 | ??3 | ??4 | ??5 | ??6 | |
| ??A | Negative sample | ??HBV?10 5 | Negative sample | ??HBV?10 5 | Negative sample | ??HBV?10 5 |
| ??B | ??HBV?10 5 | Negative sample | ??HBV?10 5 | Negative sample | ??HBV?10 5 | Negative sample |
| ??C | Negative sample | ??HBV?10 5 | Negative sample | ??HBV?10 5 | Negative sample | ??HBV?10 5 |
| ??1 | ??2 | ??3 | ??4 | ??5 | ??6 | |
| ??D | ??HBV?10 5 | Negative sample | ??HBV?10 5 | Negative sample | ??HBV?10 5 | Negative sample |
| ??E | Negative sample | ??HBV?10 5 | Negative sample | ??HBV?10 5 | Negative sample | ??HBV?10 5 |
| ??F | ??HBV?10 5 | Negative sample | ??HBV?10 5 | Negative sample | ??HBV?10 5 | Negative sample |
| ??G | Negative sample | ??HBV?10 5 | Negative sample | ??HBV?10 5 | Negative sample | ??HBV?10 5 |
| ??H | ??HBV?10 5 | Negative sample | ??HBV?10 5 | Negative sample | ??HBV?10 5 | Negative sample |
Table 2: the pcr amplification detected result of sample after contrive equipment extracts nucleic acid
| ??1 | ??2 | ??3 | ??4 | ??5 | ??6 |
| ??A | Negative | The HBV positive | Negative | The HBV positive | Negative | The HBV positive |
| ??B | The HBV positive | Negative | The HBV positive | Negative | The HBV positive | Negative |
| ??C | Negative | The HBV positive | Negative | The HBV positive | Negative | The HBV positive |
| ??D | The HBV positive | Negative | The HBV positive | Negative | The HBV positive | Negative |
| ??E | Negative | The HBV positive | Negative | The HBV positive | Negative | The HBV positive |
| ??F | The HBV positive | Negative | The HBV positive | Negative | The HBV positive | Negative |
| ??G | Negative | The HBV positive | Negative | The HBV positive | Negative | The HBV positive |
| ??H | The HBV positive | Negative | The HBV positive | Negative | The HBV positive | Negative |
Above specific examples and accompanying drawing to sample adding device of the present invention, add quadrat method and comprise that the nucleic acid test set of this sample adding device is illustrated.Yet; should be understood that; the specific examples that provides only is to illustrate for example; especially the Reference numeral in the claim only helps to understand; do not constitute restriction to the scope of protection of present invention; obviously those of ordinary skill in the art can make various corrections and change to the present invention within the scope of the invention according to the explanation of specification sheets, and for example the quadrat method that adds of operating part present embodiment is also included within the scope of protection of present invention.
Claims (13)
1. sample adding device, it comprises test tube, sample adding device shell, dispenser, stirring rod and bar magnet, it is characterized in that, comprises sample pooling zone, nucleic acid extraction district and test agent sample application zone in the described sample adding device shell, wherein,
The sample pooling zone comprises dispenser, sample liquid storage tank (11), disposable transfer pipet storage tank (12), transfer pipet disposal receptacle (13), compiles the storage tank (15) of sample liquid storage tank (14), reagent storage groove (16) and removable tote, and cuts off by the dividing plate (10) that has door (101) between sample pooling zone and the nucleic acid extraction district;
The nucleic acid extraction district comprises the storage tank (25) of storage tank (21), bar magnet cover storage tank (22), magnetic bead storage tank (24), washing lotion storage tank (23) and the second removable tote of stirring rod, bar magnet, the first removable tote;
The test agent sample application zone comprises dispenser, storage tank (41), disposable transfer pipet storage tank (42), transfer pipet disposal receptacle (43), reagent storage groove (45) and application of sample mixed solution storage tank (44);
And, the door (101) that the storage tank (21) of the first removable tote in the storage tank of the removable tote in the sample pooling zone (15) and the nucleic acid extraction district has across dividing plate (10) and adjacent, and the storage tank of the removable tote in the sample pooling zone (15) can move the test-tube stent (151) of its loading on the storage tank (21) in the first removable tote that is loaded into the nucleic acid extraction district, and the storage tank (21) of the first removable tote in the nucleic acid extraction district also can move the test-tube stent (151) of its loading and be loaded on the storage tank (15) of the removable tote in the sample pooling zone; The storage tank (25) of the second removable tote in the nucleic acid extraction district loads elutriant test-tube stent (251), and described elutriant test-tube stent (251) can be moved on the storage tank (41) in the test agent sample application zone.
2. according to the arbitrary described sample adding device of aforementioned claim, it is characterized in that, also comprise buffer zone in the described sample adding device shell, described buffer zone comprises the storage tank (31) of removable tote;
Cut off the door (201) that the storage tank (31) of the removable tote in the storage tank (25) of the second removable tote in the nucleic acid extraction district and the buffer zone has across first added partition (20) and adjacent between described buffer zone and the nucleic acid extraction district by first added partition (20) that has door (201); Cut off the door (301) that the storage tank (41) in the storage tank of the removable tote in the buffer zone (31) and the test agent sample application zone has across second added partition (30) and adjacent between described buffer zone and the test agent sample application zone by second added partition (30) that has door (301);
And, the storage tank (25) of the second removable tote in the nucleic acid extraction district can move the elutriant test-tube stent (251) of its loading and be loaded on the storage tank (31) of the removable tote in the buffer zone, and the storage tank of the removable tote in the buffer zone (15) can move the elutriant test-tube stent (251) of its loading on the storage tank (41) that is loaded in the test agent sample application zone.
3. according to the arbitrary described sample adding device of aforementioned claim, it is characterized in that in the sample pooling zone, sample liquid storage tank (11) lays respectively at the not homonymy that compiles sample liquid storage tank (14) with these two parts of storage tank (15) of removable tote.
4. according to the arbitrary described sample adding device of aforementioned claim, it is characterized in that, the nucleic acid extraction district also comprises the first auxiliary washing lotion storage tank (26) and the second auxiliary washing lotion storage tank (27), and the storage tank (25) of the storage tank of the preferred first removable tote (21), bar magnet cover storage tank (22), magnetic bead storage tank (24), washing lotion storage tank (23), the first auxiliary washing lotion storage tank (26), the second auxiliary washing lotion storage tank (27) and the second removable tote is arranged in order.
5. according to the arbitrary described sample adding device of aforementioned claim, it is characterized in that sample pooling zone, nucleic acid extraction district, buffer zone and/or test agent sample application zone comprise the gas barrier that has filter screen.
6. according to the arbitrary described sample adding device of aforementioned claim, it is characterized in that wherein said dispenser, stirring rod and bar magnet are installed in respectively on the guide rail, can carry out X, Y respectively, Z three directions move.
7. according to the arbitrary described sample adding device of aforementioned claim, it is characterized in that, wherein said dispenser is the hyperchannel dispenser that has air pressure type liquid level detector, and/or dispenser has baffle plate, and/or stirring rod has baffle plate, this baffle plate will be removed when dispenser and/or stirring rod descend, and dispenser and/or stirring rod rise to the location and will retract when mobile.
8. according to the arbitrary described sample adding device of aforementioned claim, it is characterized in that described sample adding device comprises can control dispenser, stirring rod and bar magnet operation, can control the automatic control component that storage tank that door that each dividing plate has opened and closed and can control each removable tote moves tote.
9. with the sample adding device biased sample and the compositions and methods of each claim as described above, it is characterized in that described method comprises,
(a1) in the sample pooling zone, dispenser moves and plugs the disposable transfer pipet of depositing in the disposable transfer pipet storage tank (12) successively, move and suck the sample liquid of depositing in the sample liquid storage tank (11), move and sample liquid is injected the test tube that compiles sample liquid storage tank (14), move and disposable transfer pipet is thrown aside in transfer pipet disposal receptacle (13);
(a2) then, in the sample pooling zone, dispenser moves and plugs the disposable transfer pipet of depositing in the disposable transfer pipet storage tank (12) successively, move and suck the nucleic acid lysate in the reagent storage groove (16), move and the nucleic acid lysate is injected the test tube of the test-tube stent (151) that the storage tank (15) of removable tote loads, move and disposable transfer pipet is thrown aside in transfer pipet disposal receptacle (13); In the sample pooling zone, dispenser moves and plugs the disposable transfer pipet of depositing in the disposable transfer pipet storage tank (12) successively, move and suck and compile the sample liquid of depositing in the sample liquid storage tank (14) of compiling, move and will compile the test tube that sample liquid is injected the test-tube stent (151) that the storage tank of removable tote (15) loads, move and disposable transfer pipet is thrown aside in transfer pipet disposal receptacle (13);
(a3) then, open the door (101) of the dividing plate (10) between sample pooling zone and the nucleic acid extraction district, the storage tank of the removable tote in the sample pooling zone (15) moves the test-tube stent (151) of its loading at the door (101) that the dividing plate (10) between sample pooling zone and the nucleic acid extraction district was gone up and closed to storage tank (21) in the first removable tote that is loaded into the nucleic acid extraction district, in the nucleic acid extraction district, stirring rod is moved and stirs the liquid in the test tube of described test-tube stent (151), retract stirring rod, open the door (101) of the dividing plate (10) between sample pooling zone and the nucleic acid extraction district, the storage tank (21) in the first removable tote in nucleic acid extraction district moves the door (101) that the dividing plate (10) between sample pooling zone and the nucleic acid extraction district was gone up and closed to the storage tank (15) that is loaded into the removable tote in the sample pooling zone with the test-tube stent (151) of its loading;
(a4) then, in the sample pooling zone, dispenser moves and plugs the disposable transfer pipet of depositing in the disposable transfer pipet storage tank (12) successively, move and suck in the reagent storage groove (16) in conjunction with liquid, move and will inject the test tube of the test-tube stent (151) that the storage tank (15) of removable tote loads, move and disposable transfer pipet is thrown aside in transfer pipet disposal receptacle (13) in conjunction with liquid;
(a5) then, open the door (101) of the dividing plate (10) between sample pooling zone and the nucleic acid extraction district, the storage tank of the removable tote in the sample pooling zone (15) moves the test-tube stent (151) of its loading at the door (101) that the dividing plate (10) between sample pooling zone and the nucleic acid extraction district was gone up and closed to storage tank (21) in the first removable tote that is loaded into the nucleic acid extraction district;
(b1) then, in the nucleic acid extraction district, bar magnet moves and puts the bar magnet cover of depositing in the bar magnet cover storage tank (22) successively, under magnetic state, move and absorption magnetic bead storage tank (24) in the magnetic bead deposited, under magnetic state, move and insert in the test tube of the test-tube stent (151) that the storage tank (21) of the first removable tote loads, magnetic disappears so that adsorbed magnetic bead comes off, and moves and the bar magnet cover is retracted in the bar magnet cover storage tank (22);
(b2) then, in the nucleic acid extraction district, stirring rod is moved and stirs the liquid in the test tube of the test-tube stent (151) that the storage tank (21) of the first removable tote loads, retract stirring rod;
(b3) then, in the nucleic acid extraction district, bar magnet moves and puts the bar magnet cover of depositing in the bar magnet cover storage tank (22) successively, under magnetic state, move and adsorb the magnetic bead in the test tube of the test-tube stent (151) that the storage tank (21) of the first removable tote loads, under magnetic state, move and insert in the washing lotion in the washing lotion storage tank (23), magnetic disappears so that adsorbed magnetic bead comes off, and moves and the bar magnet cover is retracted in the bar magnet cover storage tank (22);
(b4) then, in the nucleic acid extraction district, bar magnet under magnetic state, move successively and absorption washing lotion storage tank (23) in washing lotion in magnetic bead, move under magnetic state and insert in the test tube in the elutriant test-tube stent (251) that the storage tank (25) of the second removable tote loads, magnetic disappears so that adsorbed magnetic bead comes off;
(b5) in the nucleic acid extraction district, operate in or afterwards, in the test agent sample application zone, dispenser moves and plugs the disposable transfer pipet of depositing in the disposable transfer pipet storage tank (42) successively, move and suck the detection reagent in the reagent storage groove (45), move and detection reagent is injected the test tube of application of sample mixed solution storage tank (44), move and disposable transfer pipet is thrown aside in transfer pipet disposal receptacle (43);
(c) then, the elutriant test-tube stent (251) that the storage tank (25) of the second removable tote in the nucleic acid extraction district is loaded is moved on the storage tank (41) in the test agent sample application zone;
(d) then, in the test agent sample application zone, dispenser moves and plugs the disposable transfer pipet of depositing in the disposable transfer pipet storage tank (42) successively, move and suck the liquid in the test tube in the elutriant test-tube stent (251) that storage tank (41) loads, move and described liquid is injected the test tube of application of sample mixed solution storage tank (44), move and disposable transfer pipet is thrown aside in transfer pipet disposal receptacle (43).
10. biased sample according to claim 9 and compositions and methods, it is characterized in that, in step (c), open the door (201) of first added partition (20) between buffer zone and the nucleic acid extraction district, the storage tank (25) of the second removable tote in the nucleic acid extraction district moves the elutriant test-tube stent (251) of its loading and is loaded on the storage tank (31) of the removable tote in the buffer zone, close the door (201) of first added partition (20) between buffer zone and the nucleic acid extraction district, open the door (301) of second added partition (30) between buffer zone and the test agent sample application zone after leaving standstill, the storage tank of the removable tote in the buffer zone (31) moves the elutriant test-tube stent (251) of its loading on the storage tank (41) that is loaded in the test agent sample application zone, closes the door (301) of second added partition (30) between buffer zone and the test agent sample application zone.
11. the nucleic acid test set is characterized in that, described device comprises arbitrary described sample adding device and the nucleic acid tester of claim 1-8.
12. the described nucleic acid amplifier of claim 11 is characterized in that, wherein said nucleic acid tester is the PCR instrument, and the application of sample mixed solution storage tank (44) in the arbitrary described sample adding device of preferred claim 1-8 just is installed on the PCR instrument.
13. the method with the nucleic acid test set analytic sample of claim 11 or 12 is characterized in that described method comprises carries out claim 9 or 10 described biased sample and compositions and methods, detects nucleic acid with the nucleic acid tester then and analyzes.
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