CN101613663A - Full-automatic serial fluorescence PCR detection system - Google Patents
Full-automatic serial fluorescence PCR detection system Download PDFInfo
- Publication number
- CN101613663A CN101613663A CN200910041456A CN200910041456A CN101613663A CN 101613663 A CN101613663 A CN 101613663A CN 200910041456 A CN200910041456 A CN 200910041456A CN 200910041456 A CN200910041456 A CN 200910041456A CN 101613663 A CN101613663 A CN 101613663A
- Authority
- CN
- China
- Prior art keywords
- nucleic acid
- full
- reaction
- sample
- detection system
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Abstract
The invention discloses a kind of full-automatic serial fluorescence PCR detection system, comprise optophone, testing sample device, nucleic acid treatment unit, reaction unit, mechanical manipulator and guide rail, optophone is connected with the testing sample device, between testing sample device and the nucleic acid treatment unit guide rail and mechanical manipulator are set, between nucleic acid treatment unit and the reaction unit mechanical manipulator are set.Full-automatic serial fluorescence PCR detection system of the present invention is for realizing a kind of temperature variation that full-automatic PCR reaction process designs and gathering the device of fluorescence information, conventional P CR instrument has been carried out innovative improvement, changed the pattern of conventional P CR instrument batch reaction, the PCR operating mode of pipeline system has been proposed, improve the rate of utilization of PCR equipment, whole testing process realizes unmanned the intervention, fully automated, saved the labour, improved working efficiency, also reduced the influence of human factor simultaneously detected result.
Description
Technical field
The present invention relates to be used for the gene test instrument in fields such as biology, medical science, particularly can realize finishing to reaction the serial fluorescence PCR detection system of full process automatization from sample preparation.
Background technology
PCR is called for short in polymerase chain reaction (Polymerase Chain Reaction), has become the routine techniques of detection of nucleic acids.In the PCR instrument, one or more apertures that are used to place the tubule that the reactant mixed solution is housed are arranged on the thermal cycling piece.The temperature of loop blocks is in the circulation and finish the polymerase chain extension in temperature-rise period between high-temperature denatured stage of cryogenic renaturation stage and an about 90-95 degree relatively of an about 50-55 degree.Response sample on the thermal cycling piece through repeatedly " sex change ", " annealing ", " extension " stage, promptly in 90-95 degree pyroprocessing about 30 seconds, within several seconds, be cooled to the 50-55 degree then, keep after about 30 seconds and be warmed up to the 90-95 degree once more, 30-40 cycle and so forth is in the low-temperature zone in each cycle, with the fluorescent tracing thing that adds in advance in the light source activation reaction solution that adds, make it to produce fluorescence, measure this change in fluorescence, just can obtain corresponding information.
Existing P CR instrument all is concurrent working, after promptly needing to prepare enough one batch testing sample, put into the PCR treatment unit together, PCR instrument commonly used allows primary first-order equation dozens of sample mostly, such as 96 common samples, that is to say that these 96 samples will be finished in same temperature program(me).For example, the LightCycler 480 type high-throughput real-time fluorescence quantitative PCR systems of Roche Holding Ag.
In hospital or laboratory, all can use the PCR instrument at any time and detect the sample that different time is handled well.But it is because cycle of PCR is longer, waits for that a batch of reaction is finished and could begin next batch reaction, cause the PCR equipment utilization rate not high like this, and influence very much working efficiency, even frozen and can influence detected result owing to the sample needs.In addition, there is not a PCR instrument can realize not needing manual operation to the whole automatizations of finishing of reaction process at present from the processing of sample yet.Generally all need the manual handling sample, prepare the PCR reaction mixture, be put on the PCR instrument and react, also need the sample hose after the processing reaction at last.In whole PCR testing process, mixed a lot of human factors, influence the accuracy of detected result.
Summary of the invention
For overcoming above-mentioned technological deficiency, the purpose of this invention is to provide a kind of full-automatic serial fluorescence PCR detection system, improve the rate of utilization of PCR instrument greatly, and realize detecting the automatization of whole process.
For achieving the above object, adopt following technical scheme:
Full-automatic serial fluorescence PCR detection system of the present invention comprises optophone, testing sample device, nucleic acid treatment unit, reaction unit, mechanical manipulator and guide rail, described optophone is connected with described testing sample device, between described testing sample device and the described nucleic acid treatment unit described guide rail and described mechanical manipulator are set, between described nucleic acid treatment unit and the described reaction unit described mechanical manipulator are set.
Described testing sample device comprises sample disc, turning axle and stepper-motor, and described sample disc is connected with described stepper-motor by turning axle, the some holes that are used to insert sample hose of card circumference regular distribution, described sample disc upper edge.
Described nucleic acid treatment unit comprises nucleic acid process disk, rotating machinery push rod and nucleic acid treater;
Described rotating machinery push rod flexibly connects at the circle centre position of described nucleic acid process disk;
Described nucleic acid process disk comprises inner ring and outer ring, and described nucleic acid process disk is provided with 48 bank bits and 24 nucleic acid processing positions along the card circumference, and described bank bit is at described inner ring, and described nucleic acid is handled the position in described outer ring; Described outer ring is fixed, and described inner ring is rotated by computer control.Described nucleic acid treater is positioned at the periphery of described nucleic acid process disk, and is corresponding with described nucleic acid processing position.Bank bit is not limited to 48, and nucleic acid is handled the position and also is not limited to 24; This is a kind of situation in the preferred implementation of the present invention.
Described reaction unit comprises isothermal reaction dish and two temperature reaction tray.The isothermal reaction dish is meant the reaction tray of homo(io)thermism, and temperature can be selected between 37 ℃-55 ℃ and 89 ℃-95 ℃ respectively, and just the present invention is preferably 48 ℃ of reaction tray or 94 ℃ of reaction tray.
The isothermal reaction dish comprises constant temperature rotating disk, cover plate and stationary shaft; Cover plate links to each other with stationary shaft, and stationary shaft is positioned at the center of circle of isothermal reaction dish; Described isothermal reaction dish is positioned at the below of described cover plate, and described isothermal reaction rim card circumference has 20 holes that are used to bear reaction cup or pipe; There is the T-slot open card side of described cover plate; Described T-slot open card is corresponding with the hole on the described isothermal reaction dish; Two relative breach are arranged on the described cover plate.The hole that bears reaction cup or pipe is not limited to 20, and this is a kind of in the preferred implementation of the present invention.
If constant temperature is decided to be 48 ℃, then the position of two breach on its cover plate can be set according to reaction conditions, and as a preferred implementation, two breach are in relative position along circle diameter.
If constant temperature is decided to be 94 ℃, then the position of two breach on its cover plate can be set according to reaction conditions, and as a preferred implementation, the angle of two breach can be 108 degree.
Described two temperature reaction tray comprises low Winchester disk, high Winchester disk, thermofin, cover plate and stationary shaft, described low Winchester disk is positioned at the outer ring, described high Winchester disk is positioned at inner ring, cuts apart the formation lines of occlusion mutually with described thermofin between described low Winchester disk and the described high Winchester disk, and described lines of occlusion is that the gear sample is staggered; The sample pore has distributed on the same circumferential line of described lines of occlusion upper edge card circumference; Described cover plate links to each other with described stationary shaft, and described stationary shaft is positioned at the center of circle of described low Winchester disk or high Winchester disk; Described low Winchester disk and high Winchester disk are positioned at the below of described cover plate; The side perimeters of described cover plate is provided with T type channel opening card, and described T type channel opening card is corresponding with described sample pore; Sample hose is installed in the T type channel opening card of cover plate side perimeters, follows cover plate and does circumference stepwise operation and knee-action, moves to one by one the adjacent sample pore from a sample pore, has finished all after dates of sufficient temp and has just broken away from cover plate.The bottom of described sample pore is provided with the position, hole that is used to install laser diode; Sample pore on the described low Winchester disk is provided with and is used to install position, fibre-optic hole.
Full-automatic serial fluorescence PCR detection system of the present invention also comprises four hole cups, and four hole cups are positioned on the described guide rail, and described four hole cups are provided with four sample pores.As preferred implementation, described four hole cups add lysate, washings 1, washings 2 and elutriant respectively automatically in transport process.
Full-automatic serial fluorescence PCR detection system of the present invention also comprises hopper, is provided with described mechanical manipulator between described hopper and the described reaction tray.As preferred implementation, respond in the described hopper required raw material and material, such as lid, paraffin oil, reaction mixture etc.
Full-automatic serial fluorescence PCR detection system of the present invention is for realizing a kind of temperature variation that full-automatic PCR reaction process designs and gathering the device of fluorescence information.
So-called " serial " is meant each sample is entered reaction environment separately separately according to the priority order that full-automatic serial fluorescence PCR detection system of the present invention all provides an independently reaction environment for each sample; And can accomplish to finish from high temperature to low temperature repeatedly the independent collection of the information that also realized in the process in round-robin 30-40 cycle at each sample.
Compared with prior art, full-automatic serial fluorescence PCR detection system of the present invention has carried out innovative improvement to conventional P CR instrument, changed the pattern of conventional P CR instrument batch reaction, the PCR operating mode of pipeline system has been proposed, improve the rate of utilization of PCR equipment, save user's time, with survey, there are greater flexibility and suitability in the system that makes with next in realization.Owing to adopted the temperature dish of two fixed temperatures, do not adopt to produce temperature components and parts jumpy, just avoided components and parts fault because of sharply expanding with heat and contract with cold and bring for a long time, improved reliability, reduced manufacturing cost and use cost.Than adopting traditional semiconductor refrigerating, heating system can obtain higher rate of temperature change, improved reaction efficiency.Handle the processing of result even discarded reaction tubes from testing sample, whole testing process realizes unmanned the intervention, and fully automated has been saved the labour, has improved working efficiency, has also reduced the influence of human factor to detected result simultaneously.
Description of drawings
Fig. 1 is the schematic perspective view of a preferred embodiment of the testing sample device in the full-automatic serial fluorescence PCR detection system of the present invention;
Fig. 2 is the schematic top plan view of a preferred embodiment of the nucleic acid treatment unit in the full-automatic serial fluorescence PCR detection system of the present invention;
Fig. 3 is the schematic side view and the schematic top plan view of a preferred embodiment of 48 ℃ of isothermal reaction dishes in the full-automatic serial fluorescence PCR detection system of the present invention;
Fig. 4 is the schematic top plan view of a preferred embodiment of 94 ℃ of isothermal reaction dishes in the full-automatic serial fluorescence PCR detection system of the present invention;
Fig. 5 is the schematic top plan view and the schematic side view of a preferred embodiment of the two temperature reaction tray in the full-automatic serial fluorescence PCR detection system of the present invention;
Fig. 6 is the schematic top plan view and the schematic side view of a preferred embodiment of four hole cups in the full-automatic serial fluorescence PCR detection system of the present invention;
Fig. 7 is the schematic flow sheet of a preferred embodiment of the reaction process in the full-automatic serial fluorescence PCR detection system of the present invention; Wherein, Fig. 7-the 1st, full-automatic serial fluorescence PCR detection system carrying out the workflow sketch of fluorescence quantitative RT-RCR; Fig. 7-the 2nd in four hole cups, and adds the synoptic diagram of reagent with sample transfer in four hole cups; Fig. 7-the 3rd, the synoptic diagram of nucleic acid treating processes.
Embodiment
For making the present invention easier to understand,, further set forth the present invention below in conjunction with specific embodiment.Should be understood that these embodiment only to be used to the present invention is described and be not used in to limit the scope of the invention that NM concrete experimental technique in the following example carries out according to the normal experiment method usually.
Embodiment 1:
As shown in Figure 1, schematic perspective view for a preferred embodiment of the testing sample device in the full-automatic serial fluorescence PCR detection system of the present invention, as seen from the figure, the testing sample device comprises sample disc 13, turning axle 12 and stepper-motor 11, sample disc 13 is connected with stepper-motor 11 by turning axle 12, and sample disc 13 is along the some sample wells 130 that are used for reconstitute QC 15 of regular distribution on the card circumference.The staff packs testing sample into behind the sample hose 15, inserts sample well 130 again.Simultaneously stick barcode in the pipe side of sample hose 15, optophone 14 is by the response behaviour of the barcode on the identification sample hose 15 with differentiation and each testing sample pipe of tracking memory.In the present embodiment, this sample disc 13 can once be inserted 30 test tubes, and stepper-motor moved a sample position in per 30 seconds.
Embodiment 2:
As shown in Figure 2, be the vertical view of a preferred embodiment of the nucleic acid treatment unit in the full-automatic serial fluorescence PCR detection system of the present invention.This nucleic acid treatment unit comprises nucleic acid process disk 21, rotating machinery push rod 23, nucleic acid treater 24, can also comprise revolving conversion head device 22; Rotating machinery push rod 23 and revolve conversion head device 22 and flexibly connect at the circle centre position of nucleic acid process disk 21; Nucleic acid process disk 21 comprises inner ring and outer ring, and it is provided with 48 bank bits 211 and 24 nucleic acid processing positions 212 along the card circumference, and bank bit 211 is at inner ring, and nucleic acid is handled position 212 in the outer ring; The outer ring is fixed, and inner ring is rotated by computer control.Described nucleic acid treater is positioned at the periphery of described nucleic acid process disk, and is corresponding with described nucleic acid processing position.The nucleic acid treatment unit cooperates four hole cups to work together: four hole cups enter bank bit 211 by guide rail 62, rotating machinery push rod 23 moves on to a contiguous position with four hole cups then, and then, then in four hole cups, nucleic acid is handled by nucleic acid treater 24 to the nucleic acid processing position 212 that is extrapolated to the outer ring.At last, four hole cups are sent handles the position, behind the transfer nucleic acid, pushes rubbish container.
Embodiment 3
As shown in Figure 3, be the synoptic diagram of a preferred embodiment of 48 ℃ of reaction tray in the full-automatic serial fluorescence PCR detection system of the present invention.
Fig. 3-the 1st, the schematic side view of 48 ℃ of reaction tray among this embodiment, this figure shows that 48 ℃ of reaction tray comprise constant temperature rotating disk 31, cover plate 32 and stationary shaft 33, and cover plate 32 links to each other with stationary shaft 33, and stationary shaft 33 is positioned at the center of circle of 48 ℃ of reaction tray; 48 ℃ of reaction tray are positioned at the below of cover plate 32, and constant temperature rotating disk 31 is useful on the hole 311 that bears reaction cup; There is T-slot open card 321 side of cover plate 32; T-slot open card 321 is corresponding with the hole 311 on 48 ℃ of reaction tray.
Fig. 3-the 2nd, the schematic top plan view of 48 ℃ of reaction tray among this embodiment, this figure shows that constant temperature rotating disk 31 has 20 holes 311 that are used to bear reaction cup along the card circumference, 48 ℃ of constant temperature rotating disk 31 constant temperature, rotated once in per 30 seconds, and once changeed 18 degree, 10 turnbacks.Cover plate 32 upper edge circle diameters have two relative breach 322A and 322B, and reaction cup is packed into from the 322A breach, unloads from the 322B breach, follow constant temperature rotating disk 31 Rotate 180 degree, and the time is five minutes.
Embodiment 4:
Fig. 4 is the schematic top plan view of a preferred embodiment of 94 ℃ of reaction tray in the full-automatic serial fluorescence PCR detection system of the present invention.The structure of these 94 ℃ of reaction tray 4 and 48 ℃ of reaction tray is similar substantially, comprises constant temperature rotating disk, cover plate and stationary shaft, and cover plate links to each other with stationary shaft, and stationary shaft is positioned at the center of circle of 94 ℃ of reaction tray 4; 94 ℃ of reaction tray are positioned at the below of cover plate, and 94 ℃ of reaction tray 4 have 20 holes that are used to bear reaction cup along the card circumference; There is the T-slot open card side of cover plate; The T-slot open card is corresponding with the hole on described 94 ℃ of reaction tray 4; The constant temperature rotating disk has 20 holes that are used to bear reaction cup along the card circumference, and the constant temperature rotating disk rotated once in per 30 seconds, once changes 18 degree, and cover plate upper edge circumference has two breach, and reaction cup is packed into from breach 4A, and 4B unloads from breach.The difference of them part is: these 94 ℃ of reaction tray 4 constant temperature are at 94 ℃; The position difference of breach, see shown in Figure 4, the position of breach 4A and breach 4B be separated by 108 the degree, like this, the reaction times has only 3 minutes.Adjust the angle position of breach, can change heat treatment period.
Embodiment 5:
Fig. 5 is the structural representation of a preferred embodiment of the two temperature reaction tray in the full-automatic serial fluorescence PCR detection system of the present invention.Wherein, Fig. 5-the 1st, the vertical view of this two temperature reaction tray, by Fig. 5-1 as seen, this two temperature dish device comprises low Winchester disk 51, high Winchester disk 52, thermofin 53, low Winchester disk 51 and high Winchester disk 52 are two heat conductivility good metal dishes; Low Winchester disk 51 is positioned at the outer ring, and temperature can be 50-55 ℃, and high Winchester disk 52 is positioned at inner ring, and temperature can be 90-95 ℃, cuts apart the staggered lines of occlusion of formative gear sample mutually with thermofin 53 between low Winchester disk 51 and the high Winchester disk 52.Sample pore 56 has distributed on the same circumferential line of lines of occlusion upper edge card circumference.Sample pore 56 can be 40 or 80, and like this, in fact adjacent sample cup hole is positioned on the different humidity provinces.If a sample is moved into another adjacent hole from a hole, just from a humidity province to another humidity province.If this sample has moved position, 80 holes along circumference, this sample has just experienced 40 cycles that high and low temperature changes so, that is to say the whole process of having finished a PCR reaction environment.
How to realize that the sample cup moves to adjacent hole from a hole successively along circumference? this realizes by rotatable and cover plate moving up and down.Fig. 5-the 2nd, the schematic side view of this two temperature reaction tray, in conjunction with Fig. 5-2, this two temperature reaction tray also comprises cover plate 54 and stationary shaft 55 except low Winchester disk 51, high Winchester disk 52 and thermofin 53, cover plate 54 links to each other with stationary shaft 55, and stationary shaft 55 is positioned at the center of circle of low Winchester disk 51 or high Winchester disk 52; Low Winchester disk 51 and high Winchester disk 52 are positioned at the below of cover plate 54; The side perimeters of cover plate 54 is provided with 40 T type channel opening cards 541, and sample hose 59 is stuck on the cover plate 54 by T type channel opening card 541.T type channel opening card 541 is corresponding with sample pore 56; Sample hose 59 is installed in the T type channel opening card 541 of cover plate 54 side perimeters, follows cover plate 54 and does circumference stepwise operation and knee-action, and after 30 seconds, cover plate moves up, and all samples pipe 59 is extracted sample pore 56.Cover plate 54 rotates position, a hole, moves down again, and like this, all samples pipe 59 has just been changed temperature environment together.Move up and down and the stepping rotation of cover plate 54 are moved and can be driven respectively by two stepper-motors.Cover plate 54 carries sample hose and does up and down and circumferential motion, moves once in per 30 seconds, and 40 actions rotate a circle.The cover plate revolution moves position, a hole, can a new sample hose be stuck on the cover plate with mechanical manipulator, after this sample hose has been finished 40 temperature cycles, will be shifted out from cover plate.
Embodiment 6:
Fig. 6 is the synoptic diagram of a preferred embodiment of four hole cups in the full-automatic serial fluorescence PCR detection system of the present invention, and Fig. 6-1 is the schematic top plan view of four hole cups 61, and Fig. 6-2 is sectional views that four hole cups 61 are positioned over guide rail 62.Four hole cups 61 are positioned on the guide rail 62, and guide rail 62 has draw-in groove, four hole cups 61 can be fixed in the draw-in groove of guide rail, and can operation flexibly in groove.Four hole cups 61 are provided with four sample pores 611 of four sample pore 611, four hole cups 61 and put into lysate, washings 1, washings 2 and elutriant respectively.Four hole cups 61 enter stored position on the nucleic acid process disk inner ring along guide rail 62.
Embodiment 7:
Fig. 7 is the schematic flow sheet of concrete work of a preferred embodiment of full-automatic serial fluorescence PCR detection system work of the present invention.
Fig. 7-the 1st, full-automatic serial fluorescence PCR detection system carrying out the workflow sketch of fluorescence quantitative RT-RCR.Fs: after the sample preparation to be checked, the reaction mixture that contains the nucleic acid behind the purifying enters 48 ℃ of reaction tray 3.Mechanical manipulator 71C is positioned over them on 48 ℃ of reaction tray 3 grasp reaction cup 15B from hopper 72B after.Sample to be checked obtains nucleic acid behind the purifying by nucleic acid treatment unit and four hole cups 61, in last hole in four hole cups 61 of the nucleic acid behind the purifying.Four hole cups 61 are moved to by guide rail 62 and mechanical manipulator by the rotation pipettor 74B, and the nucleic acid of rotation pipettor 74B after with purifying in the four hole cups 61 is drawn among the reaction cup 15B.After rotation pipettor 74B changes head, draw reaction solution from the 73B of liquid feeding station in reaction cup 15B, wherein, liquid feeding station 73B deposits the liquid that responds in advance.Then, the mechanical manipulator 71C bowl cover that extracting cooperates with reaction cup 15B from hopper 72C again is placed on the reaction cup 15B.The above-mentioned structure that had described 48 ℃ of reaction tray 3 in detail, in brief, 48 ℃ of reaction tray 3 minutes are two-layer, and cover plate of upper layer is fixed, lower floor's rotation, per 30 seconds positions, cover plate of upper layer has material loading breach and discharging breach, and each reaction cup stopped 5 minutes at this.
Subordinate phase: reaction cup 15B transfers on 94 ℃ of reaction tray 4 from 48 ℃ of reaction tray 3.This stage action is finished by mechanical manipulator 71D.The structure of 94 ℃ of reaction sabots 4 and the structural similitude of 48 ℃ of reaction tray 3, just homo(io)thermism is at 94 ℃, and reaction cup stopped 2-3 minute at this, also entered and leave reaction unit by material loading breach and discharging breach.By adjusting the angle of two breach, can be controlled at this residence time.
Phase III: reaction cup 15B transfers on the two temperature reaction tray 5 from 94 ℃ of reaction tray 4.In the present embodiment, two temperature reaction tray 5 is provided with two.Two Winchester disks of two temperature reaction tray 5 have 40 holes, make a move in per 30 seconds, and turn over two circles and can finish 40 temperature cycles, totally 2400 seconds, promptly 40 minutes.The adjacent reaction pipe goes out one minute timed interval of result.Mechanical manipulator 71E finishes reaction cup 15B is transferred on the two temperature reaction tray 5 from 94 ℃ of reaction tray 4, and on two temperature reaction tray 5 reacted reaction cup 15B is moved to the rubbish container.
Fig. 7-the 2nd in four hole cups 61, and adds the synoptic diagram of reagent with sample transfer to four hole cups 61 in the nucleic acid treating processes.Rotation pipettor 74A draws in first hole of testing sample to four hole cup 61 from the test tube 15A that testing sample is housed.Whenever finish once, rotation pipettor 74A revolves and turn 90 degrees, and to changing " the rifle head " that head station 75A more renews, carries out the application of sample of next sample then.Lysate, washings 1, washings 2, elutriant have been deposited among the 73A of liquid feeding station respectively in advance.Added testing sample in four hole cups 61 after, the liquid among the 73A of liquid feeding station is just progressively added in four hole cups, the 61 corresponding pipes by computer program control.After sample preparation was intact, four hole cups 61 were transferred to fashionable place by guide rail 62, and wherein the nucleic acid behind the purifying is removed, and exhausted four hole cups 61 are dropped.
Fig. 7-the 3rd, the synoptic diagram of nucleic acid treating processes; Four hole cups 61 enter bank bit 211 (see figure 2)s in the nucleic acid treatment unit by guide rail 62.Rotating machine arm 23 pushes four hole cups 61 in the bank bit 211 and handles position 212.Have on the nucleic acid treater 24 can with magnet magnetic bonded guide pin bushing 241, on guide pin bushing 241 parcel one plastics tubing 242; Plastics tubing 242 can be separated with guide pin bushing 241, and works alone.In first hole, what carry out is the cracking of nucleic acid, plastics tubing 242 and guide pin bushing 241 are separated, and plastics tubing 242 stirs in first hole alone, with the liquid mixing; After 3 minutes, guide pin bushing 241 and plastics tubing 242 sockets are connected together, because put into small magnet grains in the liquid in first hole in advance, after leaving standstill, small magnet grains can be adsorbed the nucleic acid in the mixing solutions, and then can be adsorbed on the guide pin bushing 241 by this magnet, and then brings nucleic acid into second hole.In second hole, what carry out is washing.After the plastics tubing 242 that is socketed on guide pin bushing 241 enters second hole, guide pin bushing 241 is broken away from plastics tubing 242, then magnet breaks away from plastics tubing 242, thereby the nucleic acid that is adsorbed in magnet is dissolved in the washings, wash, after 3 minutes, utilize the mode in first hole, the nucleic acid after the washing is adsorbed in the 3rd hole.Solution is washings in the 3rd hole, with the same mode of second pipe.Same, at last the nucleic acid behind the wash-out is adsorbed to the 4th hole.The 4th hole is an elutriant, and such as ethanol, in ethanol eluate, nucleic acid can separate with magnet, so at last the plastics tubing 242 by being socketed with guide pin bushing 241 is with the absorption of the magnet in the solution, remaining in solution is exactly nucleic acid behind the purifying.Whole process 12 minutes.Processing makes it get back to bank bit 211 by rotating machine arm 23 after finishing.Whenever handle a cup, change a plastics tubing 242 with revolving the conversion head device.
Last institute should be noted that; above embodiment is only in order to illustrate technical scheme of the present invention but not limiting the scope of the invention; although the present invention has been done detailed description with reference to preferred embodiment; those of ordinary skill in the art is to be understood that; can make amendment or be equal to replacement technical scheme of the present invention, and not break away from the essence and the scope of technical solution of the present invention.
Claims (10)
1, full-automatic serial fluorescence PCR detection system, it is characterized in that, comprise optophone, testing sample device, nucleic acid treatment unit, reaction unit, mechanical manipulator and guide rail, described optophone is connected with described testing sample device, between described testing sample device and the described nucleic acid treatment unit described guide rail and described mechanical manipulator are set, between described nucleic acid treatment unit and the described reaction unit described mechanical manipulator are set.
2, full-automatic serial fluorescence PCR detection system according to claim 1, it is characterized in that, described testing sample device comprises sample disc, turning axle and stepper-motor, described sample disc is connected the hole that described sample disc upper edge card circumference regular distribution is used to insert sample hose by described turning axle with described stepper-motor.
3, full-automatic serial fluorescence PCR detection system according to claim 1 is characterized in that, described nucleic acid treatment unit comprises nucleic acid process disk, rotating machinery push rod and nucleic acid treater;
Described nucleic acid process disk comprises inner ring and outer ring, and described nucleic acid process disk is provided with 48 bank bits and 24 nucleic acid processing positions along the card circumference, and described bank bit is at described inner ring, and described nucleic acid is handled the position in described outer ring;
Described rotating machinery push rod flexibly connects at the circle centre position of described nucleic acid process disk;
Described nucleic acid treater is positioned at the periphery of described nucleic acid process disk, and is corresponding with described nucleic acid processing position.
4, full-automatic serial fluorescence PCR detection system according to claim 1 is characterized in that, described reaction unit comprises isothermal reaction dish and two temperature reaction tray.
5, full-automatic serial fluorescence PCR detection system according to claim 4 is characterized in that, described isothermal reaction dish comprises constant temperature rotating disk, cover plate and stationary shaft;
Described cover plate links to each other with described stationary shaft, and described stationary shaft is positioned at the center of circle of described isothermal reaction dish;
Described isothermal reaction dish is positioned at the below of described cover plate, and described isothermal reaction rim card circumference has 20 holes that are used to bear reaction cup;
There is the T-slot open card side of described cover plate; Described T-slot open card is corresponding with the hole on the described isothermal reaction dish;
Two relative breach are arranged on the described cover plate.
6, full-automatic serial fluorescence PCR detection system according to claim 4 is characterized in that, described two temperature reaction tray comprises low Winchester disk, high Winchester disk, thermofin, cover plate and stationary shaft;
Described low Winchester disk is positioned at the outer ring, and described high Winchester disk is positioned at inner ring, cuts apart the formation lines of occlusion mutually with described thermofin between described low Winchester disk and the described high Winchester disk, and described lines of occlusion is that the gear sample is staggered;
The sample pore has distributed on the same circumferential line of described lines of occlusion upper edge card circumference;
Described cover plate links to each other with described stationary shaft, and described stationary shaft is positioned at the center of circle of described low Winchester disk or high Winchester disk;
Described low Winchester disk and described high Winchester disk are positioned at the below of described cover plate.
7, full-automatic serial fluorescence PCR detection system according to claim 6 is characterized in that, the side perimeters of described cover plate is provided with T type channel opening card, and described T type channel opening card is corresponding with described sample pore.
8, full-automatic serial fluorescence PCR detection system according to claim 6 is characterized in that, the bottom of described sample pore is provided with the position, hole that is used to install laser diode; Sample pore on the described low Winchester disk is provided with and is used to install position, fibre-optic hole.
9, full-automatic serial fluorescence PCR detection system according to claim 1 is characterized in that, also comprises four hole cups, and described four hole cups are positioned on the described guide rail, and described four hole cups are provided with four sample pores.
10, full-automatic serial fluorescence PCR detection system according to claim 1 is characterized in that, also comprises hopper, between described hopper and the described reaction tray described mechanical manipulator is set.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200910041456 CN101613663B (en) | 2009-07-28 | 2009-07-28 | Full-automatic serial fluorescence PCR detection system |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200910041456 CN101613663B (en) | 2009-07-28 | 2009-07-28 | Full-automatic serial fluorescence PCR detection system |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101613663A true CN101613663A (en) | 2009-12-30 |
| CN101613663B CN101613663B (en) | 2013-01-23 |
Family
ID=41493554
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200910041456 Expired - Fee Related CN101613663B (en) | 2009-07-28 | 2009-07-28 | Full-automatic serial fluorescence PCR detection system |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101613663B (en) |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102373148A (en) * | 2010-08-13 | 2012-03-14 | 美科生物医学技术无锡有限公司 | Disc ultra high-speed real-time fluorescence quantitative PCR instrument |
| CN103614291A (en) * | 2013-11-13 | 2014-03-05 | 戴小波 | Full-automatic fluorescence quantitative gene amplification instrument |
| CN103695309A (en) * | 2013-12-19 | 2014-04-02 | 江苏金太生命科技有限公司 | Novel PCR (Polymerase Chain Reaction) instrument |
| CN104130933A (en) * | 2014-08-02 | 2014-11-05 | 张金木 | Thermostatic fluorescence PCR (polymerase chain reaction) amplifier |
| CN106885791A (en) * | 2017-03-23 | 2017-06-23 | 中国科学院合肥物质科学研究院 | Heavy metal in water automatic on-line monitoring device based on laser breakdown spectral technique |
| CN108802019A (en) * | 2017-05-02 | 2018-11-13 | 中国石油化工股份有限公司 | A kind of forecasting system and method for water of oil-gas field scaling tendency |
| CN109486655A (en) * | 2018-12-27 | 2019-03-19 | 英诺维尔智能科技（苏州）有限公司 | A kind of full-automation biological products system |
| CN110114475A (en) * | 2018-12-20 | 2019-08-09 | 广州市华南医学研究中心 | The method and its equipment of a kind of very fast nucleic acid amplification and application |
| WO2020062315A1 (en) * | 2018-09-30 | 2020-04-02 | 苏州百源基因技术有限公司 | Pcr pipeline |
| WO2021219067A1 (en) * | 2020-04-30 | 2021-11-04 | 杭州杰毅生物技术有限公司 | Nucleic acid test pipeline processing apparatus, and pipeline molecular diagnostics apparatus |
| WO2022246797A1 (en) * | 2021-05-28 | 2022-12-01 | 王锦弘 | Polymerase chain reaction device |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6703236B2 (en) * | 1990-11-29 | 2004-03-09 | Applera Corporation | Thermal cycler for automatic performance of the polymerase chain reaction with close temperature control |
| CN2612673Y (en) * | 2003-03-27 | 2004-04-21 | 上海复生生物工程研究所有限公司 | Full-automatic water-bath type gene amplification |
| EP2056114A1 (en) * | 2007-10-29 | 2009-05-06 | Koninklijke Philips Electronics N.V. | Automatic detection of infectious diseases |
| CN201530828U (en) * | 2009-07-28 | 2010-07-21 | 呼吸疾病国家重点实验室 | Full-automatic serial fluorescent PCR detecting system |
-
2009
- 2009-07-28 CN CN 200910041456 patent/CN101613663B/en not_active Expired - Fee Related
Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102373148A (en) * | 2010-08-13 | 2012-03-14 | 美科生物医学技术无锡有限公司 | Disc ultra high-speed real-time fluorescence quantitative PCR instrument |
| CN103614291A (en) * | 2013-11-13 | 2014-03-05 | 戴小波 | Full-automatic fluorescence quantitative gene amplification instrument |
| CN103695309A (en) * | 2013-12-19 | 2014-04-02 | 江苏金太生命科技有限公司 | Novel PCR (Polymerase Chain Reaction) instrument |
| CN104130933A (en) * | 2014-08-02 | 2014-11-05 | 张金木 | Thermostatic fluorescence PCR (polymerase chain reaction) amplifier |
| CN104130933B (en) * | 2014-08-02 | 2016-01-13 | 张金木 | A kind of fluorescence constant temperature PCR amplification instrument |
| CN106885791B (en) * | 2017-03-23 | 2019-04-30 | 中国科学院合肥物质科学研究院 | Heavy metal in water automatic on-line monitoring device based on laser breakdown spectral technique |
| CN106885791A (en) * | 2017-03-23 | 2017-06-23 | 中国科学院合肥物质科学研究院 | Heavy metal in water automatic on-line monitoring device based on laser breakdown spectral technique |
| CN108802019A (en) * | 2017-05-02 | 2018-11-13 | 中国石油化工股份有限公司 | A kind of forecasting system and method for water of oil-gas field scaling tendency |
| WO2020062315A1 (en) * | 2018-09-30 | 2020-04-02 | 苏州百源基因技术有限公司 | Pcr pipeline |
| CN110114475A (en) * | 2018-12-20 | 2019-08-09 | 广州市华南医学研究中心 | The method and its equipment of a kind of very fast nucleic acid amplification and application |
| WO2020124510A1 (en) * | 2018-12-20 | 2020-06-25 | 广州市华南医学研究中心 | Extremly rapid nucleic acid amplification method, apparatus and use thereof |
| CN110114475B (en) * | 2018-12-20 | 2021-06-18 | 广州市华南医学研究中心 | Method for rapid nucleic acid amplification and equipment and application thereof |
| CN109486655A (en) * | 2018-12-27 | 2019-03-19 | 英诺维尔智能科技（苏州）有限公司 | A kind of full-automation biological products system |
| WO2021219067A1 (en) * | 2020-04-30 | 2021-11-04 | 杭州杰毅生物技术有限公司 | Nucleic acid test pipeline processing apparatus, and pipeline molecular diagnostics apparatus |
| WO2022246797A1 (en) * | 2021-05-28 | 2022-12-01 | 王锦弘 | Polymerase chain reaction device |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101613663B (en) | 2013-01-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101613663B (en) | Full-automatic serial fluorescence PCR detection system | |
| CN201530828U (en) | Full-automatic serial fluorescent PCR detecting system | |
| US11828766B2 (en) | Performing antimicrobial susceptibility testing and related systems and methods | |
| AU2008222590B2 (en) | Apparatus and method for nucleic acid amplification | |
| CN207636617U (en) | It automates preanalysis sample treatment modules and container shuttle transports component | |
| CA2913698C (en) | Diagnostic systems and methods | |
| CN103003409B (en) | Method for detecting nucleic acid amplification in sample and device therefor | |
| US20020001848A1 (en) | Multi-format sample processing devices, methods and systems | |
| CN104871006B (en) | Sample processing system and the method for uniform heating slide glass | |
| CN102753978A (en) | System for conducting the identification of bacteria in biological samples | |
| JP2005507074A (en) | Method and apparatus for a high throughput sample handling process line | |
| KR20130029127A (en) | Reagent cartridge | |
| US8066943B2 (en) | Clinical analyzer having a variable cycle time and throughput | |
| US20150209789A1 (en) | Sample processing apparatus and automatic analyzing apparatus including the same | |
| CN108823088A (en) | Nucleic acid detection apparatus based on micro-fluidic chip | |
| JP5759818B2 (en) | Nucleic acid testing equipment | |
| US20090221060A1 (en) | Nucleic acid analyzer | |
| CN101801514A (en) | The thermocirculator that has the sample port of optionally opening | |
| US9593367B2 (en) | Genetic test system | |
| WO2021056005A1 (en) | Sample preparation apparatus and multi-well plate with pcr chip | |
| CN108823089A (en) | Nucleic acid detection chip based on multiple nucleic acids reaction | |
| CN110114475A (en) | The method and its equipment of a kind of very fast nucleic acid amplification and application | |
| KR101651337B1 (en) | Micro-chip for diagnosis and integrated rotary method for identification of biological pathogens using the same | |
| CN102215971A (en) | Automated analysis device with a rotating carousel for different liquid containers | |
| CN101558145A (en) | Rapid thermocycler |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| ASS | Succession or assignment of patent right |
Owner name: NATIONAL KEY EXPERIMENT OF RESPIRATORY DISEASE Free format text: FORMER OWNER: NATIONAL KEY LABORATORY OF RESPIRATORY DISEASE Effective date: 20111205 Free format text: FORMER OWNER: GUANGZHOU RUIDA BIOSCIENCE CO., LTD. Effective date: 20111205 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20111205 Address after: 510000 No. 151 Yanjiang Road, Guangzhou Applicant after: National Key Laboratory of respiratory diseases Address before: 510000 No. 151 Yanjiang Road, Guangdong, Guangzhou Applicant before: State Key Laboratory of Respiratory Diseases Co-applicant before: GUANGZHOU RIDA BIOTECH CO., LTD. |
|
| ASS | Succession or assignment of patent right |
Owner name: THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL Effective date: 20121211 Owner name: GUANGZHOU INSTITUTE OF RESPIRATORY DISEASE Free format text: FORMER OWNER: NATIONAL KEY EXPERIMENT OF RESPIRATORY DISEASE Effective date: 20121211 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20121211 Address after: 510000 No. 151 Yanjiang Road, Guangdong, Guangzhou Applicant after: Guangzhou Institute of Respiratory Disease Applicant after: The Second Affiliated Hospital of Guangzhou Medical School Address before: 510000 No. 151 Yanjiang Road, Guangdong, Guangzhou Applicant before: National Key Laboratory of respiratory diseases |
|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20130123 Termination date: 20140728 |
|
| EXPY | Termination of patent right or utility model |