CN1588004A - Two-way rotary hot wind exchanging type real time fluorescent quantitative poly enzyme chain reaction analyzer - Google Patents
Two-way rotary hot wind exchanging type real time fluorescent quantitative poly enzyme chain reaction analyzer Download PDFInfo
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- CN1588004A CN1588004A CN 200410062783 CN200410062783A CN1588004A CN 1588004 A CN1588004 A CN 1588004A CN 200410062783 CN200410062783 CN 200410062783 CN 200410062783 A CN200410062783 A CN 200410062783A CN 1588004 A CN1588004 A CN 1588004A
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Abstract
The invention discloses a dual-direction rotating hot air exchange real time fluorescence polymerase reaction analyzer, comprising tank with electric machine and prime axis inside and hot air exchange circulation device on the top. The hot air exchange circulation device consists of upper air chamber installed on upper part of tank, which has air inlet with hole on its top and the overheating protection has force fan 6 and heating unit within it. The force fan is on upper part of heating unit wherein is overheating protection, and air outlet of air inlet has extending chamber which is in tank and linked to the upper air chamber through circline enclosed air outlet. Extending chamber has temperature sensor inside and upper air chamber has controllable fan blower unit and protection device inside. The invention has properties of automatic monitoring, fast, accurate, not easy pollution, convinient operation, low price, quick popularity, suitable for the need of skeleton medical and health, epidemic prevention and plant and animal test.
Description
Technical field
The present invention is applicable to automatic, the fast quantitative analysis device of medical science, biological field polymerase chain reaction (PCR) initial target mrna concentration, more particularly, relates to a kind of two-way rotary hot wind exchanging type real time fluorescent quantitative poly enzyme chain reaction analyzer.
Background technology
PCR is that polymerase chain reaction is the abbreviation of (Polymease chain reaction), is the Protocols in Molecular Biology of the specific dna fragmentation of external at short notice a large amount of amplification.The regular-PCR instrument is the thermocirculator that earlier target gene is placed by microcomputer automatic control circuit control heating and refrigeration, finish the sex change that PCR requires, renaturation and extension, make target gene be amplified millions of times, by methods such as electrophoresis last amplified production is separated and detection after finishing pcr amplification, complex operation, analysis time is long, the method that is adopted is " end-point method " result of calculation accuracy and poor reproducibility to the initial target mrna concentration, and be difficult to realize automatical analysis, operating process very easily causes cross pollution, and analysis result once was under an embargo owing to " false positive rate " Tai Gao and is used for clinical examination.
Development in recent years the real-time fluorescence quantitative PCR technology, promptly realize special pcr amplification by temperature cycles fast, adopts the automatically quantitative analysis of special fluorescence probe and sensitive fluorescence signal reporting system realization initial target mrna concentration.Therefore, improve temperature rate and reach temperature cycles process fast, good temperature control and accurate fluorescence signal measuring system, be carry out in short-term, efficiently, the key of the PCR detection by quantitative of high specificity, favorable reproducibility.On prior art, the mode of temperature cycles employing at present has two kinds, and the one, semiconductor refrigerating heats, and the 2nd, wind is bathed.The diode mode temperature rate is slow, and analysis time is long, and the semiconductor material life-span is short.Wind is bathed mode then can improve temperature rate greatly, but its temperature homogeneity, reappearance, control accuracy are the bottleneck problems that need solve when instruments design.For improving example temperature lift-down speed, though can adopt special thin-walled glass kapillary as reaction tube, its processing technology requires high, cost an arm and a leg, and also above-mentioned apparatus structure relative complex, the instrument cost height, maintenance difficulties is big, is unsuitable for extensive popularization.
Chinese patent ZL01221110.9 discloses a kind of fluorescence quantitative polymerase chain reaction diagnostic equipment, it is to utilize control circuit fluorescence signal intensity to measured object when the control thermocirculator is to the target gene amplification to detect, because the concentration of fluorescence signal intensity and target gene is proportional relation, so fluorescence intensity just can be known the quantity or the concentration of specific gene.This measuring method is known public technology, and the present invention adopts this measuring principle.The technical scheme that this patent adopts is: whole device comprises the thermocirculator that places in the casing, fluorescence detection device, the computing machine that control circuit is outer with placing casing, the thermoelectric refrigerating unit that said thermocirculator comprises alternating temperature metal module with a socket hole, is close to the alternating temperature metal module, be located at thermoelectric refrigerating unit other heating radiator, fan and insert test tube in jack, be stamped the heat lid of band well heater on the test tube, in Re Gai, heating radiator and alternating temperature metal module, be respectively equipped with temperature sensor.The semiconductor that utilizes that this patent that hence one can see that adopts directly contacts cooling and warming with test tube, and this mode temperature rate is slow, and analysis time in the long semiconductor material life-span is short, and whole instrument needs a large amount of follow-up maintenance works, and cost of use is higher.
Application number is that 90205989.0 Chinese patent discloses a kind of by PCR DNA cloning instrument, it also is the solid well heater that adopts the semiconductor temperature difference cooling assembly to constitute, utilization produces it to its size and Orientation that changes supply current and heats or refrigeration, to the heating in turn under three kinds of constant temperature respectively of the biological sample on the objective table, thus the amplification of realization biological sample.The mode of this amplification and device have and the same deficiency of a last patent.
There is above relative deficiency at prior art, proposes the present invention.
Summary of the invention
The object of the present invention is to provide a kind of can detect fast, accurately, automatically and be difficult for to pollute, easy to operate, cheap, the two-way rotary hot wind exchanging type real time fluorescent quantitative poly enzyme chain reaction analyzer of being convenient to popularize, be suitable for unit demands such as vast primary care health, epidemic prevention, animals and plants detection.
The present invention adopts general thin-walled centrifuge tube as the PCR reaction tube, utilize the circulating air of controllable temperature to bathe realization fast, stable heat exchanging process, target gene in the reaction tube of two-way rotation is increased, finish the sex change that PCR requires, annealing, extend, simultaneously real-time detecting reactant (inspection product) and its fluorescent dye stimulated emission fluorescence intensity of reference material (the ginseng control product of concentration known), amplification curve by computer drawing reactant and reference material, and by calculating, exist (positive or negative) of target gene be can determine qualitatively, and the quantity or the concentration of original target gene in the inspection product quantitatively reported.Fast PCR amplification, real-time fluorescence detects, and qualitative, quantitative result calculates with process such as report and can finish automatically by computing machine.
For realizing above goal of the invention, technical scheme provided by the present invention is: the two-way rotary hot wind exchanging type real time fluorescent quantitative poly enzyme chain reaction analyzer, comprise casing, be provided with motor and main shaft in the casing, the casing upper end is provided with hot blast exchange cycles device, described hot blast exchange cycles device comprises the ipper air chamber that is arranged on casing top, be provided with the blast pipe of top perforate in the ipper air chamber, be provided with powerful blower fan and Heater group in the blast pipe, powerful blower fan is positioned at the upper end of Heater group, be provided with overtemperature protection system in the Heater group, the air outlet of blast pipe is provided with the amplification chamber, the amplification chamber is positioned at casing and is communicated with ipper air chamber by porous ring seal return air inlet, and the indoor temperature sensor that is provided with that increases is provided with controlled blower fan group and safety device in the ipper air chamber; The main shaft of described motor is provided with sample amplification fan rack device; Next-door neighbour's sample amplification fan rack device is provided with fluorescent quantitative detection device; Be provided with the rotation of control motor in the casing and, comprise the microprocessor of controlling the forward and reverse rotation of motor, the microprocessor of controlling microprocessor, control data collection group and the communication group of temperature by interface and the electronic-controlled installation that the casing outer computer links to each other.
The air opening end of described blast pipe is provided with the diffusing tuyere of porous wide-angle.
The indoor tubbiness return air plate that is provided with of described amplification, thus double-deck amplification chamber formed.
Described amplification outdoor is provided with synchronizing indicator and cooperates the grid shape synchronous disk of its work, and grid shape synchronous disk is installed on the main shaft.
Described sample amplification fan rack device comprises that the sample that is arranged on the indoor main shaft of amplification loads fan frame dish, evenly be provided with the sample tube hole around the sample loading fan frame dish, the upper end that sample loads fan frame dish is provided with the pressure cover that is installed on the main shaft in parallel, evenly is provided with and the corresponding elasticity compression leg in sample tube hole around the pressure cover.
Described sample loads fan frame dish and pressure cover is the aluminium profile shapes, is provided with knuckle α all around and is 10-70 ° pressure wind angle, and described test tube hole and elasticity compression leg are arranged on this pressure wind angle; Sample loads on the fan frame dish and is evenly equipped with air-permeating hole, is laid with on the pressure cover with sample and loads the air vent that the air-permeating hole on the fan frame dish is crisscross arranged.
Described knuckle α is preferably 50 °.
Described pressure cover is provided with locknut.
Cassette umbrella shape fair water cone is installed on the described locknut, and this cassette umbrella shape fair water cone spiral-lock is in the ring groove on main shaft top.
The bottom that described sample loads fan frame dish is provided with outward-dipping taper type ring-type wind-guiding tail, and tiltangle is 10-70 °.
The bottom that described sample loads fan frame dish is provided with the brute force flabellum of bleeding.
The described brute force flabellum of bleeding is straight formula, inclined or leafy chip.
Described fluorescent quantitative detection device comprises and is arranged on the outdoor light-emitting device of amplification, with light-emitting device angle β be the pick-up unit that 0-100 ° of angle is provided with, described light-emitting device comprises excitation source, exciter filter group, the convergent lens that sets gradually, pick-up unit comprises special-shaped light hurdle, dispersing lens, detection filter set, the photoelectric detector that sets gradually, special-shaped light hurdle is installed on the outer wall of amplification chamber, other refrigeration cover and the refrigerator group of being provided with of photoelectric detector.
The angle β of described light-emitting device and pick-up unit is preferably 90 °.
Described excitation source is tungsten lamp, high-brightness LED light emitting diode or the laser of wavelength coverage 350-800nm, described exciter filter group and detection filter set are the optical filter of the wavelength coverage 350-800nm of 1-6 bit pattern use, and described photoelectric detector is photomultiplier, phototriode or photoelectric cell.
Described refrigerator group is to cover for directly contacting the semiconductor chilling plate of refrigeration with refrigeration.
The present invention produce beneficial effect have following some:
1, adopt New Cycle hot blast bath pattern to realize temperature cycles, adopted air channel brand-new, that optimize design simultaneously, temperature rate is enhanced about more than once than traditional semiconductor refrigerating pattern, has realized quick, stable temperature cycles process.
2, designed unique bi-directional variable speed rotary sample amplification fan frame system, the heat interchange and the heat transfer process of sample solution in air, amplification tube wall and the pipe have been promoted greatly, significantly improved the actual lifting temperature rate of amplification inner reaction tube sample, make temperature homogeneity and accuracy in amplification chamber and the amplification pipe reach optimum condition simultaneously, and make sample more even, thereby guaranteed the height reappearance of amplification efficiency with mixing of amplifing reagent and fluorescent dye.
3, designed the 90 degree vertical optical path systems of fluoroscopic examination the best, compared, can significantly reduce the interference of excitation source and source of stray light, improved signal to noise ratio (S/N ratio) greatly detection signal with traditional parallel light path system.
4, this instrument adopts general, inexpensive thin-walled centrifuge tube can realize the requirement of rapid quantitative pcr analysis as the sample amplification pipe, can reduce running cost, simultaneously by above-mentioned brand-new design simplification apparatus structure, guaranteeing to greatly reduce instrument cost on the reliable performance basis of instrument, therefore the two-way rotary hot wind exchanging type real time fluorescent quantitative poly enzyme chain reaction analyzer of the present invention's design is convenient to popularize, is suitable for promote, and particularly can satisfy the demand of units such as vast primary care health, epidemic prevention, animals and plants detection.
In sum, the present invention adopts the bi-directional variable speed circulating air to bathe mode, the effectively design by a series of innovations, solved the key technical problem of quantitative fluorescent PCR, simplified apparatus structure simultaneously, greatly reduced instrument cost, had and use widely and promotion prospect.
Description of drawings
1. Fig. 1 is a general function block diagram of the present invention
2. Fig. 2 is a structural representation of the present invention
3. Fig. 3 loads fan shelf structure synoptic diagram for sample
4. Fig. 4 is the vertical view of Fig. 3
5. Fig. 5 is the upward view of Fig. 3
6. Fig. 6 is a porous ring seal return air inlet structural representation
7. Fig. 7 is the vertical view of Fig. 6
Wherein, the direction of arrow is the wind flow direction among Fig. 2, and four-headed arrow represents that motor can two-wayly rotate.Among the figure, hot blast exchange cycles device A; sample amplification fan rack device B; fluorescent quantitative detection device C; electronic-controlled installation D; computing machine E; casing 1; motor 2; main shaft 3; ipper air chamber 4; blast pipe 5; powerful blower fan 6; Heater group 7; overtemperature protection system 8; amplification chamber 9; porous ring seal return air inlet 10; temperature sensor 11; controlled blower fan group 12; safety device 13; the porous wide-angle tuyere 14 that looses; tubbiness return air plate 15; synchronizing indicator 16; grid shape synchronous disk 17; sample loads fan frame dish 18; sample tube hole 19; pressure cover 20; elasticity compression leg 21; press wind angle 22; air-permeating hole 23; air vent 24; locknut 25; cassette umbrella shape fair water cone 26; wind-guiding tail 27; the flabellum 28 of bleeding; light-emitting device 29; pick-up unit 30; excitation source 31; exciter filter group 32; convergent lens 33; special-shaped light hurdle 34; dispersing lens 35; detect filter set 36; photoelectric detector 37; refrigeration cover 38; refrigerator group 39; sample tube 40
Embodiment
Below in conjunction with accompanying drawing the present invention is further specifically described:
Fig. 1 is a general function block diagram of the present invention, shown in Fig. 1 frame of broken lines, analyser of the present invention comprises five devices---hot blast exchange cycles device A, sample amplification fan rack device B, fluorescent quantitative detection device C, electronic system device D and external computer device E, hot blast exchange cycles device A is arranged on the upper end of casing 1, sample amplification fan rack device B is arranged on the main shaft 3 of motor 2, the sample amplification that fluorescent quantitative detection device C is arranged in the casing 1 is fanned by the rack device B, electronic-controlled installation D links to each other with casing 1 outer computer E by interface, and electronic-controlled installation D is just comprising control motor 2, the microprocessor of reverse rotation, the microprocessor of control temperature, the microprocessor of control data collection group and communication group.
More than function of realizing of each device as follows:
Hot blast exchange cycles device A, its function one is to create a good temperature environment for the target gene amplification, under specific temperature environment, finishes the required a plurality of temperature cycle work of sex change, annealing, extension of target gene amplification, reaches the purpose of amplification.Its function two is for the heat interchange circulation provides one to meet the structure that the air fluid circulation requires, and realizes quick, stable heat interchange and thermal cycle.
Sample amplification fan rack device B, its function has four: one, carrying analytic sample guarantees consistance and the locational requirement of sample tube in hot blast bath environment.Two, guarantee that each sample tube can work under tight condition, do not leak sample so that contaminated environment.Three, the huge draft that produces in the high speed rotating is full of the amplification chamber with hot-air in the utmost point short time, and makes it evenly to make the temperature homogeneity of sample reach optimum condition, to guarantee the height repeatability of amplification efficiency.Four, under the effect of bi-directional variable speed rotation, promote the heat interchange and the heat transfer process of interior sample solution of pipe and amplification tube wall and amplification chamber air, improve the actual temperature rate of amplification inner reaction tube sample, the interior sample of sample tube is mixed fully with fluorescent dye, and make the temperature of sample in the pipe more even.
The function of fluorescent quantitative detection device C mainly contains five aspects, and one, provide a strong stable excitation source for instrument.Two, provide a fluorescence detection device that sensitivity is suitable for instrument.Three, the 90 degree vertical optical path systems of the required the best of fluoroscopic examination are provided for instrument, have compared, can significantly reduce excitation source and source of stray light, significantly improved signal to noise ratio (S/N ratio) the interference of detection signal with traditional parallel light path system.Four, for providing, instrument can satisfy the user easily to exciting or detect the demand that wavelength combinations changes in the filter set of a plurality of position groupings installations.Five, the working environment of a constant temperature is provided for pick-up unit.
The function of electronic-controlled installation is to satisfy the required control function demand of analytical instrument and keep in communication with the external computer device to get in touch.
The function of external computer device is to provide a work control basic platform for analysis by the operating analysis special software.
Fig. 2 is a structural representation of the present invention; as shown in Figure 2; the upper end of casing 1 is provided with ipper air chamber 4; be provided with the blast pipe 5 of top perforate in the ipper air chamber 4; blast pipe 5 is communicated with casing 1; its inside is provided with powerful blower fan 6 and Heater group 7; powerful blower fan 6 is positioned at the upper end of Heater group 7; this Heater group 7 is provided with overtemperature protection system 8, and the air opening end of blast pipe 5 extends in the amplification chamber 9 in the casing 1, and the porous wide-angle tuyere 14 that looses is installed; ipper air chamber 4 links to each other with amplification chamber 9 by porous ring seal return air inlet 10; amplification is provided with tubbiness return air plate 15 in the chamber 9, makes this amplification chamber form double-deck amplification chamber, is beneficial to circulating of air-flow more.Be provided with controlled blower fan group 12 in the ipper air chamber 4 and safety device 13 is installed, evenly distribute in the amplification chamber 9 4-6 group temperature sensor 11 is installed, so that the temperature in the amplification chamber 9 is monitored in real time, the signal of the microprocessor collecting temperature sensor 11 in the electronic-controlled installation D also transfers to outer computer E, makes it carry out the temperature control heating by the parameter that expection is provided with.
The diffusing tuyere 14 of porous wide-angle is horn-like, and is provided with orifice plate in the upper end, is connected with the endpiece of blast pipe 5, makes the hot blast of blowing over Heater group 7 enter amplification chamber 9 more uniformly.
The bilayer amplification chamber 9 that is provided with tubbiness return air plate 15 has guaranteed that the hot blast that enters amplification chamber 9 gets back to ipper air chamber 4 by porous ring seal return air inlet 10 faster, and controlled blower fan group 12 has been accelerated the shuttling movement of this hot blast.The hot blast thermal losses that interlayer has guaranteed to enter amplification chamber 9 reaches minimum, thereby the heat interchange and the heat transfer process of the chamber air that can promote to increase improve the actual temperature rate of amplification inner reaction tube sample.
But the bi-directional variable speed motor rotating 2 by electronic-controlled installation D control is arranged on outside the amplification chamber 9, and the main shaft 3 of motor 2 extends in the amplification chamber 9, is provided with grid shape synchronous disk 17 on the main shaft 3 between motor 2 and the amplification chamber 9, and synchronizing indicator 16 cooperates its work.Amplification is provided with sample amplification fan rack device B in the chamber 9, comprises that the sample that is arranged on the main shaft 3 loads fan frame dish 18, and it is the aluminium profile shapes that sample loads fan frame dish 18, is provided with the pressure wind angle 22 of knuckle α between 10-70 ° all around, and present embodiment is got α=50 °.Press wind angle 22 to be provided with the sample tube hole 19 that to insert sample tube 40.It is the aluminium profile shapes that this sample loads fan frame dish 18, is evenly equipped with air-permeating hole 23 on it, and the bottom is provided with outward-dipping taper type ring-type wind-guiding tail 27, and sample loads and fan frame dish 18 bottom level angle theta between 10-70 °, and present embodiment is got θ=50 °.Sample loads brute force that fan frame dish 18 bottoms the are provided with one flabellum 28 of bleeding, and this brute force flabellum 32 of bleeding can be straight formula, inclined or leafy chip, and present embodiment adopts straight formula, and adopts and be weldingly fixed on dish 18 and the wind-guiding tail 27 (as shown in Fig. 3,4,5).
The upper end that sample loads fan frame dish 18 is provided with the pressure cover 20 that is installed on the main shaft 3 in parallel, pressure cover 20 is the aluminium profile shapes, it is identical that its structure, shape and sample load fan frame dish 18, difference is that knuckle 22 is provided with and sample tube hole 19 corresponding elasticity compression legs 21, is evenly equipped with on the pressure cover 20 with sample and loads the air vent 24 that the air-permeating hole 23 on the fan frame dish 18 shifts to install.
Pressure cover 20 is provided with locknut 25, and locknut 25 upper ends are the cassette umbrella shape fair water cone 26 that is installed on the main shaft 3 terminal ring grooves, are provided with spring steel ball in this cassette umbrella shape fair water cone 26, directly is buckled in interior the getting final product of ring groove of main shaft 3 ends during installation.
During practical application, at first will be placed on the sample tube hole 19 at the sample tube 40 that detected reactant is housed, cover pressure cover 20, locknut 25 backspins are locked, then cassette umbrella shape fair water cone 26 is pressed, made that the elasticity compression leg 21 around the pressure cover 20 is pushed down sample tube 40 to guarantee the tightness of sample tube 40.The sample tube hole of the present invention's design is fit to the test tube of multiple models such as 0.1ml, 0.2ml, 0.5ml.Owing to press the feature inclined design at wind angle 22, sample tube 40 after installing tilts with the axis direction of main shaft 3, such effect is can larger area to accept the irradiation of excitation source for the sample in amplification later test tube, has further guaranteed the ageing and accuracy of pick-up unit testing result; The test tube of Qing Xieing has quickened the circulation of hot blast just as one by one small-sized fans leaf simultaneously.
The hot blast of blowing down from blast pipe 5 is full of amplification chamber 9 uniformly after the porous wide-angle looses tuyere 14, the cassette umbrella shape fair water cone 26 under it make sample carrying fan frame dish 18 that hot blast disperses faster around, pass through sample tube 40 fast.Bleeder vent 23 on air vent 24 on the pressure disc 20 and the sample that is crisscross arranged with it carrying fan frame dish 18 makes and passes through the more even of sample tube 40, has avoided hot blast to concentrate on a certain zone, may be to the destruction of sample tube 40 thereby produce.Outward-dipping taper type ring-type wind-guiding tail 27 below the sample carrying fan frame dish 18 and the powerful flabellum 28 of bleeding make discharges through the skin of bilayer amplification chamber 9 fast by the hot blast of sample tube 40, and pass through porous ring seal return air inlet 10 and get back to upper chamber 4, the controlled blower fan group 12 in the upper chamber 4 has been quickened this circulation.The structure of porous ring seal return air inlet 10 is shown in Fig. 6,7.
By above heated air circulation device A and amplification chamber 9 special structural design, make hot-air be full of the amplification chamber, and make it evenly to make the temperature homogeneity of sample reach optimum condition, to guarantee the height repeatability of amplification efficiency in the utmost point short time.And guarantee that each sample tube can work under tight condition, do not leak sample so that contaminated environment.Can also be under the effect of bi-directional variable speed rotation, promote the heat interchange and the heat transfer process of interior sample solution of pipe and amplification tube wall and amplification chamber air, improve the actual temperature rate of amplification inner reaction tube sample, the interior sample of sample tube is mixed fully with fluorescent dye, and make the temperature of sample in the pipe more even.
Outside in amplification chamber 9 is provided with fluorescent quantitative detection device C, comprise the light-emitting device of forming by excitation source 31, exciter filter group 32, convergent lens 33 29, pick-up unit 30 is β=0-100 ° of angle setting with light-emitting device 29, and present embodiment adopts 90 ° of angles vertically to be provided with.This pick-up unit 30 comprises special-shaped light hurdle 34, dispersing lens 35, detection filter set 36, the photoelectric detector of placing successively 37, wherein, special-shaped light hurdle 34 is installed on the outer wall of amplification chamber 9, photoelectric detector 37 other refrigeration cover 38 and the refrigerator groups 39 that are provided with direct contact refrigeration.
Photoelectric detector 37 of the present invention is photomultiplier, phototriode or photoelectric cell, and exciter filter group 17 is used wavelength coverage 350-800nm for the 1-6 bit pattern.Detect filter set 36 and use wavelength coverage 350-800nm for the 1-6 bit pattern.Excitation source 31 is tungsten lamp, high-brightness LED light emitting diode or the laser of wavelength coverage 350-800nm.Present embodiment adopts the tungsten lamp of wavelength coverage 350-800nm as excitation source, adopt photomultiplier as photoelectric detector, adopt the optical filter combination of 4 wavelength coverage 400-750nm to use, adopt the optical filter combination of 4 wavelength coverage 400-750nm to use as detecting filter set as the exciter filter group.
Under enlightenment of the present invention, can be in casing 1 a plurality of positions be provided with sample tube hole 19 in sample tube 40 corresponding light-emitting device 29 and pick-up units 30.
The course of work of analytical instrument of the present invention is: under the control of the built-in multidigit microprocessor of external computer E and the electronic-controlled installation D that is attached thereto by interface according to the running parameter that sets in advance, analytic sample is carried out hot blast bathe circulation, bathe in the circulation at hot blast, powerful blower fan 6 in the instrument top blast pipe 5 blows into temperature control heating in the Heater group 7 with air at room temperature, blow to uniformly in the double layer return air amplification chamber 9 by the diffusing tuyere 28 of porous wide-angle through heated hot air heated stream, sample amplification fan rack device B timesharing bi-directional variable speed rotation heat of stirring airflow in amplification chamber 9, the powerful exhaust fan 28 that loads fan frame dish 18 bottoms at sample cooperates the very big draft of formation with pressing wind angle 22, make the hot-air high velocity stream through sample, bringing the sample heat heats up sample, and sample loads the cassette umbrella shape fair water cone 36 at fan frame 18 tops and the special construction of double-deck amplification chamber 9, good loop is provided for the high-speed and high-temperature airflow, shown in the direction of arrow among Fig. 2.Bathe circulation time at cold wind, the controlled blower fan group 12 of 4 afterbodys is opened work during windward, quickens air draft, and amplification chamber 9 interior temperature are reduced fast to reach the demand of amplification.Again in the heating and cooling process, the rotation of sample amplification fan frame 18 timesharing bi-directional variable speed drives sample tube internal reaction liquid and forms vorticity, promote interior solution of pipe and tube wall and pipe outer air that heat interchange takes place rapidly and transmit, finish cyclic processes such as the required distortion of gene magnification, annealing, extension fast with heat.In amplification procedure, fluorescent quantitative detection device C with 29 one-tenth an angle of 90 degrees of light-emitting device, detect the fluorescent dye strength in the sample tube 40 turn over special-shaped light hurdle 34 fast by demand, and curve plotting, obtain the fluorescent dye intensity curve (amplification curve) of one group of example reaction thing (inspection product) and reference material (the ginseng control product of concentration known) through the work in 40 cycles too, after handling calculating, obtain the Ct value of reference material and the working curve of concentration and Ct (fitting formula and linearly dependent coefficient), draw the initial copy number (concentration) of example reaction thing (inspection product) again after as calculated, and positive result of determination.If what adopt is many fluorescent dyes system, instrument can be by being provided with the fluorescence curve that can detect 2-6 different wave length simultaneously.
Those skilled in the art can carry out other changes and modification after reading patent specification of the present invention, but these changes and modification are all within the claim protection domain that the present invention is awaited the reply.
Claims (16)
1. two-way rotary hot wind exchanging type real time fluorescent quantitative poly enzyme chain reaction analyzer, comprise casing (1), be provided with motor (2) and main shaft (3) in the casing (1), casing (1) upper end is provided with hot blast exchange cycles device (A), it is characterized in that: described hot blast exchange cycles device (A) comprises the ipper air chamber (4) that is arranged on casing (1) top, be provided with the blast pipe (5) of top perforate in the ipper air chamber (4), be provided with powerful blower fan (6) and Heater group (7) in the blast pipe (5), powerful blower fan (6) is positioned at the upper end of Heater group (7), be provided with overtemperature protection system (8) in the Heater group (7), the air outlet of blast pipe (5) is provided with amplification chamber (9), amplification chamber (9) is positioned at casing (1) and is communicated with ipper air chamber (4) by porous ring seal return air inlet (10), be provided with temperature sensor (11) in the amplification chamber (9), be provided with controlled blower fan group (12) and safety device (13) in the ipper air chamber (4);
The main shaft (3) of described motor (2) is provided with sample amplification fan rack device (B); Next-door neighbour's sample amplification fan rack device (B) is provided with fluorescent quantitative detection device (C);
Be provided with control motor (2) rotation in the casing (1) and, comprise the microprocessor of controlling the forward and reverse rotation of motor (2), the microprocessor of controlling microprocessor, control data collection group and the communication group of temperature by the electronic-controlled installation (D) that interface links to each other with casing (1) outer computer (E).
2. two-way rotary hot wind exchanging type real time fluorescent quantitative poly enzyme chain reaction analyzer according to claim 1 is characterized in that: the air opening end of described blast pipe (5) is provided with the diffusing tuyere (14) of porous wide-angle.
3. two-way rotary hot wind exchanging type real time fluorescent quantitative poly enzyme chain reaction analyzer according to claim 1 is characterized in that: be provided with tubbiness return air plate (15) in the described amplification chamber (9), thereby form double-deck amplification chamber.
4. two-way rotary hot wind exchanging type real time fluorescent quantitative poly enzyme chain reaction analyzer according to claim 1, it is characterized in that: outside, described amplification chamber (9) is provided with synchronizing indicator (16) and cooperates the grid shape synchronous disk (17) of its work, and grid shape synchronous disk (17) is installed on the main shaft (3).
5. two-way rotary hot wind exchanging type real time fluorescent quantitative poly enzyme chain reaction analyzer according to claim 1, it is characterized in that: described sample amplification fan rack device (B) comprises that the sample on the main shaft (3) that is arranged in the amplification chamber (9) loads fan frame dish (18), evenly be provided with sample tube hole (19) around the sample loading fan frame dish (18), the upper end that sample loads fan frame dish (18) is provided with the pressure cover (20) that is installed on the main shaft (3) in parallel, evenly is provided with and sample tube hole (19) corresponding elasticity compression legs (21) around the pressure cover (20).
6. two-way rotary hot wind exchanging type real time fluorescent quantitative poly enzyme chain reaction analyzer according to claim 5, it is characterized in that: described sample loads fan frame dish (18) and pressure cover (20) is the aluminium profile shapes, be provided with knuckle α all around and be 10-70 ° pressure wind angle (22), described test tube hole (19) is arranged on this pressure wind angle (22) with elasticity compression leg (21); Sample loads on the fan frame dish (18) and is evenly equipped with air-permeating hole (23), is laid with on the pressure cover (20) with sample and loads the air vent (24) that the air-permeating hole (23) on the fan frame dish (18) is crisscross arranged.
7. two-way rotary hot wind exchanging type real time fluorescent quantitative poly enzyme chain reaction analyzer according to claim 6 is characterized in that: described knuckle α is 50 °.
8. two-way rotary hot wind exchanging type real time fluorescent quantitative poly enzyme chain reaction analyzer according to claim 6 is characterized in that: described pressure cover (20) is provided with locknut (25).
9. two-way rotary hot wind exchanging type real time fluorescent quantitative poly enzyme chain reaction analyzer according to claim 8, it is characterized in that: cassette umbrella shape fair water cone (26) is installed on the described locknut (25), and this cassette umbrella shape fair water cone (26) spiral-lock is in the ring groove on main shaft (3) top.
10. according to the described two-way rotary hot wind exchanging type real time fluorescent quantitative poly enzyme chain reaction analyzer of each claim of 5-9, it is characterized in that: the bottom that described sample loads fan frame dish (18) is provided with outward-dipping taper type ring-type wind-guiding tail (27), and tiltangle is 10-70 °.
11. according to the described two-way rotary hot wind exchanging type real time fluorescent quantitative poly enzyme chain reaction analyzer of each claim of 5-9, it is characterized in that: the bottom that described sample loads fan frame dish (18) is provided with the brute force flabellum (28) of bleeding.
12. two-way rotary hot wind exchanging type real time fluorescent quantitative poly enzyme chain reaction analyzer according to claim 11 is characterized in that: the described brute force flabellum (28) of bleeding is straight formula, inclined or leafy chip.
13. two-way rotary hot wind exchanging type real time fluorescent quantitative poly enzyme chain reaction analyzer according to claim 1, it is characterized in that: described fluorescent quantitative detection device (C) comprises the light-emitting device (29) that is arranged on outside the amplification chamber (9), with light-emitting device (29) angle β be the pick-up unit (30) that 0-100 ° of angle is provided with, described light-emitting device (29) comprises the excitation source (31) that sets gradually, exciter filter group (32), convergent lens (33), pick-up unit (30) comprises the special-shaped light hurdle (34) that sets gradually, dispersing lens (35), detect filter set (36), photoelectric detector (37), special-shaped light hurdle (34) is installed on the outer wall of amplification chamber (9), other refrigeration cover (38) and the refrigerator group (39) of being provided with of photoelectric detector (37).
14. two-way rotary hot wind exchanging type real time fluorescent quantitative poly chain according to claim 13 is anti-, answers analyser, it is characterized in that: light-emitting device (29) is 90 ° with the angle β of pick-up unit (30).
15. two-way rotary hot wind exchanging type real time fluorescent quantitative poly enzyme chain reaction analyzer according to claim 13, it is characterized in that: described excitation source (31) is the tungsten lamp of wavelength coverage 350-800nm, high-brightness LED light emitting diode or laser, described exciter filter group (32) and detection filter set (36) are the optical filter of the wavelength coverage 350-800nm of 1-6 bit pattern use, and described photoelectric detector (37) is photomultiplier, phototriode or photoelectric cell.
16. two-way rotary hot wind exchanging type real time fluorescent quantitative poly enzyme chain reaction analyzer according to claim 13 is characterized in that: described refrigerator group (39) is to cover (38) for directly contacting the semiconductor chilling plate of refrigeration with refrigeration.
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| Application Number | Priority Date | Filing Date | Title |
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| CN 200410062783 CN1281944C (en) | 2004-07-09 | 2004-07-09 | Two-way rotary hot wind exchanging type real time fluorescent quantitative poly enzyme chain reaction analyzer |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410062783 CN1281944C (en) | 2004-07-09 | 2004-07-09 | Two-way rotary hot wind exchanging type real time fluorescent quantitative poly enzyme chain reaction analyzer |
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| Publication Number | Publication Date |
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| CN1588004A true CN1588004A (en) | 2005-03-02 |
| CN1281944C CN1281944C (en) | 2006-10-25 |
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| CN 200410062783 Expired - Fee Related CN1281944C (en) | 2004-07-09 | 2004-07-09 | Two-way rotary hot wind exchanging type real time fluorescent quantitative poly enzyme chain reaction analyzer |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013131274A1 (en) * | 2012-03-09 | 2013-09-12 | 瑞基海洋生物科技股份有限公司 | Device and method for controlling thermal convection velocity of biochemical reaction |
| CN104136914A (en) * | 2011-12-15 | 2014-11-05 | 海因生命科学股份有限公司 | A device for optically measuring fluorescence of nucleic acids in test samples and use of the device |
| CN107058090A (en) * | 2017-04-27 | 2017-08-18 | 滨江华康（北京）生物科技有限公司 | A kind of real-time fluorescence quantitative PCR gene magnification detector |
| CN110951604A (en) * | 2019-12-31 | 2020-04-03 | 广州源起健康科技有限公司 | Rotating device for real-time quantitative gene amplification fluorescence detection |
-
2004
- 2004-07-09 CN CN 200410062783 patent/CN1281944C/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104136914A (en) * | 2011-12-15 | 2014-11-05 | 海因生命科学股份有限公司 | A device for optically measuring fluorescence of nucleic acids in test samples and use of the device |
| CN104136914B (en) * | 2011-12-15 | 2016-08-24 | 海因生命科学股份有限公司 | The device of fluorescence and the use of this device for optical measurement sample amplifying nucleic acid |
| WO2013131274A1 (en) * | 2012-03-09 | 2013-09-12 | 瑞基海洋生物科技股份有限公司 | Device and method for controlling thermal convection velocity of biochemical reaction |
| CN107058090A (en) * | 2017-04-27 | 2017-08-18 | 滨江华康（北京）生物科技有限公司 | A kind of real-time fluorescence quantitative PCR gene magnification detector |
| CN110951604A (en) * | 2019-12-31 | 2020-04-03 | 广州源起健康科技有限公司 | Rotating device for real-time quantitative gene amplification fluorescence detection |
| CN110951604B (en) * | 2019-12-31 | 2023-07-21 | 广州源起健康科技有限公司 | Rotary device for real-time quantitative gene amplification fluorescence detection |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1281944C (en) | 2006-10-25 |
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