CN2482080Y - Fluorescent quantitative polymerase chain reaction diagnostic apparatus - Google Patents
Fluorescent quantitative polymerase chain reaction diagnostic apparatus Download PDFInfo
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- CN2482080Y CN2482080Y CN 01221110 CN01221110U CN2482080Y CN 2482080 Y CN2482080 Y CN 2482080Y CN 01221110 CN01221110 CN 01221110 CN 01221110 U CN01221110 U CN 01221110U CN 2482080 Y CN2482080 Y CN 2482080Y
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Abstract
The utility model relates to a fluorescence quantitative polymerase chain reaction diagnostic instrument, which comprises a heat circulating device a fluorescence detection device, a controlling circuit all of which are arranged in a box body, and a computer arranged outside the box body. The utility model is controlled through the computer and a microprocessor, while the heat circulating device implements the increase to a target gene, the fluorescence intensity of the fluorescence detection device to a tested thing and a standard thing is detected, the fluorescence intensity of the tested thing is compared with the fluorescence intensity of the standard thing through a computer software, because the target gene intensity is proportional to the fluorescence intensity, the real-time quantitative detection of the target gene concentration can be automatically and rapidly realized, because the whole detection process is carried on in a fully enclosed test tube, the tested thing can not be environmentally polluted.
Description
The utility model is a device biological, that medical domain is used for quantitative test polymerase chain reaction (PCR) initial target mrna concentration.
Before the utility model is made, also there is not the instrument of full-automatic detection by quantitative polymerase chain reaction initial target gene.Traditional method is the thermocirculator that earlier target gene is placed by microcomputer automatic control circuit control heating and refrigeration, finishes sex change, renaturation and extension that PCR requires, makes target gene be amplified millions of times.And then amplified production is carried out gel electrophoresis judge whether to have the target gene that desire detects.Because the efficient of pcr amplification is very high, the target gene of minute quantity and a large amount of target genes are increased simultaneously finally to reach capacity, result when electrophoresis detection is the same, therefore this method can only obtain the qualitative results whether specific gene exists, and can't be to its quantity or concentration detection by quantitative, this in addition method is owing to need detections sample is shifted in operating process, so is subjected to the pollution of environment easily and causes result inaccurate (false positive and false negative).Also there is complicated operating process simultaneously, shortcoming such as detection time is long.
The purpose of this utility model provides a kind of detection by quantitative that can finish automatically, fast target gene, and is not subjected to the fluorescence quantitative polymerase chain reaction diagnostic equipment of environmental pollution in whole testing process.
Technical solution of the present utility model is: utilize control circuit fluorescence signal intensity to tested reactant 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 (quantity in the unit volume) of specific gene.
Whole device comprises the thermocirculator that places in the casing, fluorescence detection device, control circuit and place the outer computing machine of 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;
Fluorescence detection device comprises that the socket hole with on the metal module that is positioned at alternating temperature metal module below is parallel, and drive the synchronous band move around by stepper motor, be fixed on synchronously with on housing, housing is provided with position transducer, the housing top is porose, be provided with lens down successively from topped hole, with axis be 45 ° of inclination angles two to dichronic mirror, cross bar and photomultiplier with axis normal, cross bar is through relative two walls of housing, this cross bar moves around with housing, location-plate bump on its two end and the casing is done horizontal slip, on cross bar, be provided with the optical filter of some different wave lengths, be provided with excitation source above in housing, being positioned at cross bar, optical filter, the light that lens, light source send is through optical filter, the lens level shines two on dichronic mirror;
Control circuit comprises the microprocessor that links to each other with the peripheral hardware computing machine by interface, to convert the A/D converter that digital signal is defeated by microprocessor from the output signal of alternating temperature metal module temperature sensor and photomultiplier to respectively, input end links to each other with microprocessor, the solid-state commutation bridge that is used to make voltage commutation that output is electrically connected with thermoelectric refrigerating unit, input end links to each other with temperature sensor, output covers temperature-control circuit and the Switching Power Supply that well heater links to each other with fan with heat, Switching Power Supply links to each other with microprocessor, its output voltage is defeated by solid-state commutation bridge respectively, temperature-control circuit and stepper motor driver, the drive signal input stepper motor driver of microprocessor output, the output signal input microprocessor of position transducer.
Below in conjunction with accompanying drawing the utility model is described in further detail:
Fig. 1 is the synoptic diagram of fluorescence quantitative polymerase chain reaction diagnostic equipment of the present utility model;
Fig. 2 is the thermocirculator synoptic diagram;
Fig. 3 is the A-A cut-open view of Fig. 1;
Fig. 4 is that control circuit constitutes block diagram;
Fig. 5 is a kind of physical circuit example of solid-state commutation bridge of control circuit;
Fig. 6 is the physical circuit example of the Switching Power Supply in the control circuit;
Fig. 7 is the physical circuit example of temperature-control circuit.
With reference to Fig. 1, fluorescence quantitative polymerase chain reaction diagnostic equipment of the present utility model comprises the thermocirculator I that places in the casing V, fluorescence detection device II, control circuit III and place the outer Computer I V of casing, its fluorescence detection device is positioned at the below of thermocirculator.The structure of thermocirculator and fluorescence detection device is seen Fig. 2, shown in Figure 3.
The thermoelectric refrigerating unit 3 that said thermocirculator I comprises alternating temperature metal module 4 with a socket hole 21, is close to alternating temperature metal module 4, be located at thermoelectric refrigerating unit 3 other heating radiator 2, fan 1 and insert being used in jack 21 and hold the test tube 5 of reactant, above test tube 5, be stamped the heat lid 6 of the band well heater that prevents the evaporation of reaction in test tube thing, in heat lid 6, heating radiator 2, be respectively equipped with temperature sensor 7, in alternating temperature metal module 4, be provided with temperature sensor 8; During use, hold the standard reaction thing of concentration known, in all the other test tubes, hold tested reactant, and in reactant, add fluorescer with several test tubes.
Fluorescence detection device II comprises that the socket hole 21 with on the metal module that is positioned at alternating temperature metal module 4 belows is parallel, and by stepper motor 17 drive move around be with 16 synchronously, be fixed on synchronously with the housing 22 on 16, housing is provided with position transducer 36, housing top porose 19, be provided with lens 13 down successively from topped hole, with axis be 45 ° of inclination angles two to dichronic mirror 12, cross bar 20 and photomultiplier 15 with axis normal, cross bar 20 is through relative two walls of housing, this cross bar moves around with housing, location-plate 18 bumps on its two end and the casing are done horizontal slip, on cross bar, be provided with the optical filter 14 of some different wave lengths, be provided with excitation source 9 above in housing, being positioned at cross bar, optical filter 10, lens 11.Usually, use the LED excitation source, the exciting light that excitation source 9 sends is filtered into the monochromatic light of specific wavelength by optical filter 10, be focused into directional light by lens 11 again and shine two on dichronic mirror 12, the monochromatic light of specific wavelength reflects the bottom that scioptics 13 focus on test tube 5 through two to dichronic mirror, make invisible spectro reactant send the fluorescence of specific wavelength, fluorescence scioptics 13 are radiated at two on dichronic mirror 12, wavelength passes two to dichronic mirror 12 greater than the fluorescence of certain value, be filtered into the monochromatic light of specific wavelength through optical filter 14, received by photomultiplier 15 and become electric signal and be defeated by control circuit.Stepper motor 17 forward and reverse rotations drive is with 16 rectilinear motions synchronously, thereby make be fixed on synchronously with on the housing with optical detection device move around, fluorescence to each test tube detects one by one, switch optical filter 14 by location-plate 18 bump cross bars 20 simultaneously, be used to filter the fluorescence of different wave length.
Thermocirculator and fluorescence detection device are by control circuit control carrying out work and information transmission.
Control circuit is seen shown in Figure 4, it comprises the microprocessor 25 that links to each other with peripheral hardware Computer I V by interface 24, to convert the A/D converter 26 that digital signal is defeated by microprocessor 25 from the output signal of temperature sensor 8 and photomultiplier 15 to respectively, 27, input end links to each other with microprocessor, the solid-state commutation bridge 28 that is used to make voltage commutation that output is electrically connected with thermoelectric refrigerating unit 3, input end links to each other with temperature sensor 7, output covers temperature-control circuit 30 and the Switching Power Supply 29 that well heater links to each other with fan 1 with heat, Switching Power Supply 29 links to each other with microprocessor 25, its output voltage is defeated by solid-state commutation bridge 28 respectively, temperature-control circuit 30 and stepper motor driver 34, the drive signal input stepper motor driver 34 of microprocessor output, the output signal input microprocessor of position transducer 36.
During work, the command signal that Computer I V spreads out of converts control signal to through microprocessor 25, microprocessor control signal is defeated by solid-state commutation bridge 28 on the one hand, voltage from Switching Power Supply 29 is commutated and regulate by it, heat supply electricity refrigerator 3 heating or refrigeration or constant temperature, the temperature requirement of control pcr amplification, be defeated by stepper motor driver 34 on the other hand, the control step motor is realized reciprocating type straight line stepping, finishes successively all fluorescence intensity detection of (comprising standard reaction thing and tested reactant) test-tube reaction thing under different wave length.Simultaneously, the temperature signal of temperature sensor 8 outputs in the thermocirculator and the electric signal of 15 outputs of the photomultiplier in the fluorescence detection device are respectively by A/D converter 26,27 convert digital signal to, and the position signalling of position transducer 36 outputs all is transferred to computing machine through microprocessor, through after the software processing, sending on the one hand steering order is defeated by microprocessor and converts control signal to, on the other hand the tested reactant imported in the amplification procedure and the fluorescence intensity of standard reaction thing are made comparisons, be processed into visual and understandable curve or chart and show, realize detection by quantitative target gene at display.Temperature-control circuit 30 in the control circuit receives the temperature signal of temperature sensor 7 outputs, and heat lid well heater is connected in its output, and fan is so that heat lid and environment temperature are controlled at required scope all the time.
Fig. 5 is a kind of physical circuit example of solid-state commutation bridge of control circuit;
In this example, solid-state commutation bridge is to constitute TE among the figure by two IR2110 integrated packages and MOSFET pipe Q1～Q4
+, TE
-End links to each other with thermoelectric refrigerating unit, and COOL, HEAT, PWM end are the cpu i/f signals that comes from microprocessor, and this circuit has been realized contactless electronic commutation.
Fig. 6 is the physical circuit example of the Switching Power Supply in the control circuit.
In this example, AC power behind rectifying and wave-filtering and fuse, enters the DC to DC converter module PH300F of isolation, at HV through the TVS protection of pipe
+, HV
-The higher DC voltage of end output safety characteristic.DC voltage offers the driving that solid-state commutation bridge is used for thermoelectric refrigerating unit, also offers step motor drive voltage, provides 12VDC to temperature-control circuit simultaneously, is used for the temperature control of heat lid, heating radiator.Link to each other with the power supply control interface of microprocessor between the CNT among the PH300F and SG.
Fig. 7 is the physical circuit example of temperature-control circuit.
Heat covers the temperature signal of temperature sensor and goes into from SEN6, OUT6 termination, and with in integrated circuit U5 (TLC2202), carry out amplification ratio by the temperature set points of resistance R 7, R8 decision, drive the on off state of Q5, the heat lid heating film that butts up against the HOTL10 end is realized temperature automatically controlled.In like manner, the temperature signal of temperature sensor is gone into from SEN7, OUT7 termination on the heating radiator, and with in integrated circuit U6 (TLC2202), carry out amplification ratio by the temperature set points of resistance R i2 decision, drive the on off state of Q6, control is connected to the fan action of FAN end, the temperature of realization heating radiator is controlled automatically, and U3 and U4 are constant current sources, and it has played the effect of stable dut temperature signal.
Microprocessor in the control circuit can be used 80C196.Interface can adopt RS232.
Fluorescence quantitative polymerase chain reaction diagnostic equipment of the present utility model, utilize computing machine, microprocessor control, when thermocirculator is implemented amplification to target gene, adopt fluorescence detection device that the fluorescence intensity of checking matter and reference material is detected, and the fluorescence intensity of checking matter and reference material is made comparisons by computer software, because target gene intensity and fluorescence intensity are proportional, therefore can be automatic, realize fast the real-time quantitative of target gene concentration is detected, because whole testing process is to carry out in airtight fully test tube, and the fluorescer that adds test tube does not pollute reactant, and therefore detecting thing can not be subjected to environmental pollution.
Claims (2)
1. fluorescence quantitative polymerase chain reaction diagnostic equipment, it is characterized in that it comprises the thermocirculator [I] that places in the casing, fluorescence detection device [II], control circuit [III] and place the outer computing machine [IV] of casing, said thermocirculator [I] comprises the alternating temperature metal module [4] with a socket hole [21], the thermoelectric refrigerating unit of being close to alternating temperature metal module [4] [3], be located at the other heating radiator [2] of thermoelectric refrigerating unit [3], fan [1] and insert test tube [5] in jack [21], be stamped the heat lid [6] of band well heater on the test tube [5], in Re Gai [6], be respectively equipped with temperature sensor [7] in the heating radiator [2], in alternating temperature metal module [4], be provided with temperature sensor [8];
Fluorescence detection device [II] comprises that the socket hole [21] with on the metal module that is positioned at alternating temperature metal module [4] below is parallel, and drive the synchronous band [16] move around by stepper motor [17], be fixed on the housing [22] on the synchronous band [16], housing is provided with position transducer [36], housing top porose [19], be provided with lens [13] down successively from topped hole, with axis be 45 ° of inclination angles two to dichronic mirror [12], cross bar [20] and photomultiplier [15] with axis normal, cross bar [20] is through relative two walls of housing, this cross bar moves around with housing, location-plate [18] bump on its two end and the casing is done horizontal slip, on cross bar, be provided with the optical filter [14] of some different wave lengths, be provided with excitation source [9] above in housing, being positioned at cross bar, optical filter [10], lens [11], the light that light source sends is through optical filter [10], lens [11] level shines two on dichronic mirror [12];
Control circuit [III] comprises the microprocessor [25] that links to each other with peripheral hardware computing machine [IV] by interface [24], to convert the A/D converter [26 that digital signal is defeated by microprocessor [25] to from the output signal of temperature sensor [8] and photomultiplier [15] respectively, 27], input end links to each other with microprocessor, the solid-state commutation bridge [28] that is used to make voltage commutation that output is electrically connected with thermoelectric refrigerating unit [3], input end links to each other with temperature sensor [7], output covers temperature-control circuit [30] and the Switching Power Supply [29] that well heater links to each other with fan [1] with heat, Switching Power Supply [29] links to each other with microprocessor [25], its output voltage is defeated by solid-state commutation bridge [28] respectively, temperature-control circuit [30] and stepper motor driver [34], the drive signal input stepper motor driver [34] of microprocessor output, the output signal input microprocessor of position transducer [36].
2. by the described fluorescence quantitative polymerase chain reaction diagnostic equipment of claim 1, it is characterized in that said solid-state commutation bridge is to be made of two IR2110 integrated packages and MOSFET pipe [Q1～Q4].
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 01221110 CN2482080Y (en) | 2001-04-12 | 2001-04-12 | Fluorescent quantitative polymerase chain reaction diagnostic apparatus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 01221110 CN2482080Y (en) | 2001-04-12 | 2001-04-12 | Fluorescent quantitative polymerase chain reaction diagnostic apparatus |
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| CN2482080Y true CN2482080Y (en) | 2002-03-13 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN 01221110 Expired - Fee Related CN2482080Y (en) | 2001-04-12 | 2001-04-12 | Fluorescent quantitative polymerase chain reaction diagnostic apparatus |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100378227C (en) * | 2003-05-08 | 2008-04-02 | 生物-拉德实验室公司 | Systems and methods for fluorescence detection with a movable detection module |
| CN105733939A (en) * | 2014-11-14 | 2016-07-06 | 财团法人工业技术研究院 | Constant temperature rotary device |
| CN106834110A (en) * | 2015-12-04 | 2017-06-13 | 北京倍肯华业科技发展有限公司 | A kind of isothermal PCR detector and its detection method |
| 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 |
| CN109929751A (en) * | 2019-04-23 | 2019-06-25 | 苏州合惠生物科技有限公司 | A kind of novel fluorescence quantitative augmentation detection instrument |
-
2001
- 2001-04-12 CN CN 01221110 patent/CN2482080Y/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100378227C (en) * | 2003-05-08 | 2008-04-02 | 生物-拉德实验室公司 | Systems and methods for fluorescence detection with a movable detection module |
| CN105733939A (en) * | 2014-11-14 | 2016-07-06 | 财团法人工业技术研究院 | Constant temperature rotary device |
| CN105733939B (en) * | 2014-11-14 | 2018-03-23 | 财团法人工业技术研究院 | Constant temperature rotary device |
| CN106834110A (en) * | 2015-12-04 | 2017-06-13 | 北京倍肯华业科技发展有限公司 | A kind of isothermal PCR detector and its detection method |
| 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 |
| CN109929751A (en) * | 2019-04-23 | 2019-06-25 | 苏州合惠生物科技有限公司 | A kind of novel fluorescence quantitative augmentation detection instrument |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C56 | Change in the name or address of the patentee |
Owner name: HANGZHOU BORI TECHNOLOGY CO., LTD. Free format text: FORMER NAME OR ADDRESS: DAHE THERMOMAGNETIC ELECTRONIC CO., LTD., HANGZHOU |
|
| CP03 | Change of name, title or address |
Address after: 310012, No. 90, East Software Park, Wensanlu Road, Xihu District, Hangzhou Patentee after: Hangzhou BIOER Technology Co., Ltd. Address before: Hangzhou City, Zhejiang province 310005 111 South Jinjiang building 9F-B Patentee before: Hangzhou Daiwa Thermomagnetic Electronic Co., Ltd. |
|
| C19 | Lapse of patent right due to non-payment of the annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |