CN110628607A - Efficient constant-temperature PCR device suitable for emergency field application and implementation method - Google Patents
Efficient constant-temperature PCR device suitable for emergency field application and implementation method Download PDFInfo
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Abstract
The invention relates to a high-efficiency constant-temperature PCR device suitable for the field application of emergency and a realization method thereof, wherein a shell of the device is internally provided with a machine core, the top surface of the shell is provided with a screen and a hot cover positioned on the machine core; the movement comprises a fluorescence sensor, an X-axis movement mechanism, a Y-axis movement mechanism, a main control board, a sample seat, a support frame and a support bottom plate, wherein the main control board, the support frame and the X-axis movement mechanism are arranged on the support bottom plate, the sample seat is arranged on the support frame, the Y-axis movement mechanism is vertically arranged on the X-axis movement mechanism, and the fluorescence sensor is arranged on the Y-axis movement mechanism. All detection hole sites on the same sample seat can be independently started or stopped for detection, and the function of simultaneously starting or stopping detection of all the detection hole sites is kept.
Description
Technical Field
The invention relates to a PCR gene amplification instrument, in particular to a high-efficiency constant-temperature PCR device suitable for field application of an emergency and an implementation method.
Background
The PCR gene amplification instrument is an instrument and equipment for amplifying specific DNA by utilizing a PCR (Polymerase chain reaction) technology, is an extremely important tool for molecular biology research, is mainly applied to the subdivision research directions of pathogen detection, drug efficacy assessment, tumor gene detection, gene expression research, transgene research, Single Nucleotide Polymorphism (SNP) and mutation analysis and the like, and has wide application in the life science fields of food detection, clinical examination, disease control, inspection and quarantine, scientific research laboratories, food safety, cosmetic detection, environmental sanitation and the like.
At present, all detection holes on the same sample seat of the common constant temperature PCR on the market can only be started or stopped for detection at the same time, and the detection result can be obtained and a report can be formed after the constant temperature PCR is generally operated for about 1 hour. If the instrument is operated for detection, when a new burst sample is to be detected in the detection process, the next detection can be carried out only after the current detection is finished, and the working efficiency of the instrument is low in the life science fields which need field application and can quickly and timely form a detection report, such as food detection, clinical examination, disease control, inspection and quarantine and the like. Especially for sudden safety events, such as epidemic situations occurring in a farm, if a detection report cannot be formed quickly and timely, huge loss may be caused to users.
For small-batch samples to be detected generated by an emergency, a constant temperature PCR (polymerase chain reaction) with which all detection hole sites on the same sample seat can be independently started or stopped needs to be developed, corresponding detection hole sites are independently started according to the number of the samples to be detected, and the started detection hole sites enter a detection state. And if the hole site in the detection state is detected completely or the detection of the abnormal hole site is stopped artificially, the hole site is restored to the idle state again. As long as the sample seat has the idle hole sites, the instrument can continuously work to detect the newly generated sudden samples to be detected, the detection efficiency is greatly improved, and a feasible, efficient and rapid detection method is provided for the life science fields needing field application, such as food detection, clinical examination, disease control, inspection and quarantine, and the like.
Disclosure of Invention
The invention provides a high-efficiency constant-temperature PCR device suitable for field application of an emergency and an implementation method thereof, aiming at improving the detection efficiency of an instrument and ensuring the rapid and timely formation of a detection report.
In order to achieve the purpose, the technical scheme of the invention is as follows:
a high-efficiency constant-temperature PCR device suitable for emergency field application comprises a machine core, a screen, a hot cover and a shell, wherein the machine core is arranged in the shell, the screen is arranged on the top surface of the shell, and the hot cover is arranged on the machine core; the movement comprises a fluorescence sensor, an X-axis movement mechanism, a Y-axis movement mechanism, a main control board, a sample seat, a support frame and a support base plate, wherein the main control board, the support frame and the X-axis movement mechanism are installed on the support base plate, the sample seat is installed on the support frame, the Y-axis movement mechanism is vertically installed on the X-axis movement mechanism, the fluorescence sensor is installed on the Y-axis movement mechanism, the X-axis movement mechanism is matched with the Y-axis movement mechanism to drive the fluorescence sensor to move to the position below any detection hole of the sample seat, and then the fluorescence amplification detection of a sample to be detected is realized by controlling the fluorescence sensor by the main control.
Further, the fluorescence sensor employs a silicon photodiode as a fluorescence receiving sensor.
Furthermore, the X-axis movement mechanism and the Y-axis movement mechanism are both accurately positioned by adopting a mechanism that a stepping motor drives a screw rod pair to be matched with a microswitch.
Furthermore, the main control board adopts an embedded ARM as a main control chip.
Furthermore, the movement comprises two sample seats, each sample seat is provided with eight detection hole sites for supporting the placement of a common eight-connecting pipe; a thermal cover is arranged above each sample seat to ensure that the temperature of the sample seat can be constant.
Further, the supporting frame and the supporting bottom plate are both made of aluminum plates, and black anodic oxidation treatment is carried out.
Furthermore, the screen adopts a seven-inch industrial android screen,
further, the shell is manufactured by adopting D printing.
A high-efficiency constant-temperature PCR implementation method suitable for emergency field application adopts a high-efficiency constant-temperature PCR device suitable for emergency field application, and comprises the following steps:
1) initially, all detection hole sites of the sample seat are in an idle state;
2) selecting all detection hole sites of the same sample seat to start and stop detection or single detection hole site to start and stop detection at the same time through a human-computer interaction interface of a screen;
3) for the field application of the emergency, when a small quantity of samples to be detected need to be detected, the corresponding detection hole sites are selected to be independently started according to the quantity of the samples to be detected, the started detection hole sites enter a detection state, and meanwhile, the screen sends the positions of the independently started detection hole sites to the main control board;
4) after the main control board receives the position of the single starting hole site sent by the screen, the X-axis movement mechanism and the Y-axis movement mechanism are controlled to drive the fluorescence sensor to move to the position below a target detection hole site of the sample seat, and then the fluorescence sensor is controlled to realize fluorescence amplification detection of a sample to be detected;
5) and (3) controlling the fluorescence sensor to sequentially complete the fluorescence amplification detection of all samples to be detected according to the target hole site position sequence by the main control board, and then repeating the detection of the steps 4) to 5) after the next detection period is reached.
Further: 1) if the hole site detection time in the detection state is up, or the detected abnormal hole site is stopped manually through the start-stop detection of a single detected hole site on the screen man-machine interaction interface, the hole site is restored to the idle state again; 2) if a new sudden sample to be detected exists and the sample seat is provided with a detection hole site in an idle state, the new sample to be detected is placed into the idle detection hole site, and then the new detection hole site is independently started, so that the hole site in the detection state cannot be influenced, and the high-efficiency constant-temperature PCR device does not need to be interrupted; after the next new detection period comes, adding a target hole position sequence to the current newly started detection hole position, and repeating the detection of the steps 4) -5) by the high-efficiency constant-temperature PCR device; 3) all detection hole sites of same sample seat all can select single detection hole site to open and stop to detect and be in the hole site detection time of detection state and arrive or can resume idle state again after the artificial unusual stop, as long as there is idle hole site on the sample seat, high-efficient constant temperature PCR device can continuous uninterrupted duty, remains all detection hole sites simultaneously and opens the function that stops the detection, realizes the high-efficient constant temperature amplification of high-efficient constant temperature PCR device through this kind of mode and detects.
The invention has the beneficial effects that:
all detection hole sites on the same sample seat can be independently started or stopped for detection, and the hole sites which are started to be in a detection state can be restored to an idle state again under the condition that the detection time is reached or the detection is artificially abnormally stopped, so that continuous uninterrupted work can be realized by the instrument as long as the idle hole sites exist on the sample seat, the detection efficiency and the timeliness are greatly improved, and the device is very suitable for the field application occasions of emergency events requiring rapid and timely formation of detection reports. And meanwhile, the function of starting and stopping detection of all detection hole sites simultaneously is reserved, and the application of the instrument is more flexible.
Drawings
FIG. 1 is a schematic diagram of a high-efficiency isothermal PCR device suitable for emergency field application according to the present invention;
fig. 2 is a core module assembly view;
FIG. 3 is a diagram of a human-computer interaction interface;
in the figure: 1. the system comprises a machine core, 2 a screen, 3 a thermal cover, 4 a shell, 11 a fluorescent sensor, 12 an X-axis motion mechanism, 13 a Y-axis motion mechanism, 14 a main control board, 15 a sample seat, 16 a support frame, 17 a support bottom board, 21 all detection hole sites are simultaneously started and stopped for detection, 22 single detection hole site is started and stopped for detection, and 23 a curve is displayed.
Detailed Description
An embodiment of the present invention will be described in further detail with reference to the accompanying drawings, which are provided for illustration purposes only and are not intended to limit the scope of the invention.
As shown in fig. 1 to 3, a high-efficiency constant-temperature PCR device suitable for emergency field application comprises a movement 1, a screen 2, a thermal cover 3 and a housing 4, wherein the movement shown in fig. 2 further comprises a fluorescence sensor 11, an X-axis movement mechanism 12, a Y-axis movement mechanism 13, a main control board 14, a sample holder 15, a support frame 16 and a support bottom board 17. The movement 1 is arranged in the shell 4, the screen 2 and the thermal cover 3 are arranged on the shell 4, the main control board 14, the support frame 16 and the X-axis movement mechanism 12 are arranged on the support bottom board 17, the sample seat 15 is arranged on the support frame 16, the Y-axis movement mechanism 13 is vertically arranged on the X-axis movement mechanism 12, and the fluorescence sensor 11 is arranged on the Y-axis movement mechanism 13.
The X-axis motion mechanism 12 is matched with the Y-axis motion mechanism 13 to drive the fluorescence sensor 11 to move to the position below any detection hole of the sample seat 15 along the X-axis and the Y-axis, and then the main control board 14 controls the fluorescence sensor 11 to realize fluorescence amplification detection of a sample to be detected.
As shown in fig. 2, the fluorescence sensor 11 of the movement 1 adopts a silicon photodiode as a fluorescence receiving sensor, the X-axis movement mechanism 12 and the Y-axis movement mechanism 13 both adopt a stepping motor to cooperate with a microswitch, so as to realize accurate positioning, the main control board 14 adopts an embedded ARM as a main control chip, the movement 1 comprises 2 sample seats 15, each sample seat has 8 detection hole sites, support common 8 connecting pipes to be placed, the support frame 16 and the support base plate 17 both adopt aluminum plates, and black anodic oxidation treatment is performed. Screen 2 adopts 7 cun industry android screens, and every sample seat top all has a hot lid 3 to guarantee that the temperature of sample seat 15 can be invariable, the shell adopts 3D to print.
Each sample holder 15 can independently control the temperature and is divided into two temperature areas, namely an area A and an area B, 8 hole sites of each temperature area are sequentially defined as A1-A8 and B1-B8, each hole site can carry out single detection hole site start-stop detection 22 or all detection hole sites can simultaneously start-stop detection 21 through a human-computer interaction interface of the screen 2, and an amplification curve in the detection process can be displayed on a curve display 23 interface.
The invention relates to a high-efficiency constant-temperature PCR implementation method suitable for field application of emergency events, which comprises the following steps:
(1) all the detection well sites of the sample holder 15 can be in two states: an idle state or a detection state. Initially, all the detection hole sites of the sample holder 15 are in an idle state;
(2) all detection hole sites of the same sample holder 15 can be selected to be simultaneously started and stopped for detection 21 or a single detection hole site can be simultaneously started and stopped for detection 22 through a man-machine interaction interface of a screen 2 shown in fig. 3;
(3) for the field application of the emergency, when a small batch of samples to be detected need to be detected, the corresponding detection hole sites can be independently started according to the number of the samples to be detected. The started detection hole sites enter a detection state, and meanwhile, the screen 2 sends the positions of the detection hole sites which are started independently to the main control board 14;
(4) after the main control board 14 receives the position of the single start hole sent by the screen 2, firstly, the X-axis movement mechanism 12 and the Y-axis movement mechanism 13 are controlled to drive the fluorescence sensor 11 to move to the position below the target detection hole of the sample holder 15, and then the fluorescence sensor 11 is controlled to realize the fluorescence amplification detection of the sample to be detected;
(5) the main control board 14 controls the fluorescence sensor 11 to sequentially complete fluorescence amplification detection of all samples to be detected according to the target hole site position sequence, and then repeats the detection of the steps (4) to (5) after the next detection period is reached;
(6) if the hole site detection time in the detection state is up (namely the detection is finished), or the detection of the abnormal hole site is stopped manually through the start-stop detection 22 of a single detection hole site on the human-computer interaction interface of the screen 2, the hole site is restored to the idle state again;
(7) if a new sudden sample to be detected exists and the sample seat is provided with a detection hole site in an idle state, the new sample to be detected is placed into the idle detection hole site, then the new detection hole site is independently started, the hole site in the detection state cannot be influenced, and the operation of the instrument does not need to be interrupted. After waiting for the next new detection period, adding a target hole position sequence to the current newly started detection hole position, and repeating the detection in the steps (4) to (5) by the instrument;
(8) all detection hole sites of the same sample holder 15 can be selected to start and stop the detection 22 at a single detection hole site, and the hole site detection time in the detection state reaches or can be recovered to the idle state again after being artificially abnormally stopped, so long as the idle hole sites exist on the sample holder, the instrument can continuously and uninterruptedly work, and simultaneously, the functions of starting, stopping and detecting all the detection hole sites simultaneously are retained, and the efficient constant-temperature amplification detection of the instrument is realized through the mode.
Initially, all the test wells of the sample holder 15 are idle. Assuming that 3 samples to be detected are provided, putting a sample 1 into an A1 hole, putting a sample 2 into an A6 hole, putting a sample 3 into a B3 hole, then respectively starting the A1, A6 and B3 hole sites, and setting the detection time to be 45min, then the main control board 14 controls the X-axis movement mechanism 12 and the Y-axis movement mechanism 13 to drive the fluorescence sensor 11 to move to the positions below the target detection hole sites of the two sample seats 15 according to the target hole site position sequence (A1, A6 and B3), and further controls the fluorescence sensor 11 to sequentially complete the fluorescence amplification detection of all the samples to be detected.
And if 3 new samples to be detected are generated after 10min, the operation of the instrument is not required to be interrupted, the new samples to be detected are respectively placed into the A2 hole site, the B1 hole site and the B8 hole site, then the corresponding detection hole sites are started, the detection time is set to be 45min, and then the newly started A2 hole site, B1 hole site and B8 hole site are added into the target hole site position sequence. The main control board 14 controls the X-axis movement mechanism 12 and the Y-axis movement mechanism 13 to drive the fluorescence sensor 11 to move to the lower part of the target detection hole sites of the two sample seats 15 according to the new target hole site position sequence (a1, a2, a6, B1, B3, B8), and further controls the fluorescence sensor 11 to sequentially complete the fluorescence amplification detection of all samples to be detected.
And recovering the idle state after the detection of the A1, A6 and B3 hole sites is finished, and continuously carrying out fluorescence amplification detection on the A2, B1 and B8 hole sites.
If the hole site detection of the sample B1 to be detected at the 2 nd time is found to be abnormal in the detection process, stopping the B1 hole site with abnormal detection manually through the single detection hole site start-stop detection 22 of the human-computer interaction interface of the screen 2, restoring the B1 hole site to the idle state again, and continuing the fluorescence amplification detection on the A2 hole site and the B8 hole site.
For small batches of samples to be detected generated by emergencies, as long as the sample seat has idle hole sites, the instrument can continuously work, and can quickly and timely form a detection report, so that the detection efficiency is greatly improved, and the practical, feasible, efficient and quick detection method is provided for the life science fields needing field application, such as food detection, clinical inspection, disease control, inspection and quarantine, and the like.
Claims (10)
1. A high-efficiency constant-temperature PCR device suitable for emergency field application comprises a machine core (1), a screen (2), a heat cover (3) and a shell (4), and is characterized in that: a machine core (1) is arranged in the shell (4), a screen (2) is arranged on the top surface of the shell (4), and a hot cover (3) positioned on the machine core (1) is arranged; the movement (1) comprises a fluorescence sensor (11), an X-axis movement mechanism (12), a Y-axis movement mechanism (13), a main control board (14), a sample seat (15), a support frame (16) and a support base plate (17), wherein the main control board (14), the support frame (16) and the X-axis movement mechanism (12) are installed on the support base plate (17), the sample seat (15) is installed on the support frame (16), the Y-axis movement mechanism (13) is vertically installed on the X-axis movement mechanism (12), the fluorescence sensor (11) is installed on the Y-axis movement mechanism (13), the X-axis movement mechanism (12) is matched with the Y-axis movement mechanism (13) to drive the fluorescence sensor (11) to move to the position below any detection hole of the sample seat (15), and then the fluorescence amplification detection of a sample to be detected is realized by the main control plate (14) and the fluorescence sensor (11).
2. The PCR device of claim 1, wherein the PCR device is suitable for field application in emergency, and comprises: the fluorescence sensor (11) adopts a silicon photodiode as a fluorescence receiving sensor.
3. The PCR device of claim 1, wherein the PCR device is suitable for field application in emergency, and comprises: the X-axis movement mechanism (12) and the Y-axis movement mechanism (13) are both accurately positioned by adopting a mechanism that a stepping motor drives a screw rod pair to be matched with a microswitch.
4. The PCR device of claim 1, wherein the PCR device is suitable for field application in emergency, and comprises: the main control board (14) adopts an embedded ARM as a main control chip.
5. The PCR device of claim 1, wherein the PCR device is suitable for field application in emergency, and comprises: the movement (1) comprises two sample seats (15), each sample seat (15) is provided with eight detection hole sites for supporting the placement of common eight-connecting pipes; a thermal cover (3) is arranged above each sample holder (15) to ensure that the temperature of the sample holder (15) can be constant.
6. The PCR device of claim 1, wherein the PCR device is suitable for field application in emergency, and comprises: the supporting frame (16) and the supporting bottom plate (17) are both made of aluminum plates, and black anodic oxidation treatment is carried out.
7. The PCR device of claim 1, wherein the PCR device is suitable for field application in emergency, and comprises: the screen (2) adopts a 7-inch industrial android screen.
8. The PCR device of claim 1, wherein the PCR device is suitable for field application in emergency, and comprises: the shell (4) is manufactured by 3D printing.
9. A method for realizing high-efficiency constant-temperature PCR suitable for emergency field application, which adopts the high-efficiency constant-temperature PCR device suitable for emergency field application of any one of claims 1 to 8, and is characterized in that: the method comprises the following steps:
1) at the beginning, all the detection hole sites of the sample seat (15) are in an idle state;
2) selecting all detection hole sites of the same sample seat (15) through a human-computer interaction interface of a screen (2) to start and stop detection (21) or start and stop detection (22) of a single detection hole site;
3) for the field application of emergency, when a small batch of samples to be detected need to be detected, the corresponding detection hole sites are selected to be independently started according to the number of the samples to be detected, the started detection hole sites enter a detection state, and meanwhile, the screen (2) sends the positions of the independently started detection hole sites to the main control board (14);
4) after the main control board (14) receives the position of the single starting hole position sent by the screen (2), firstly controlling the X-axis movement mechanism (12) and the Y-axis movement mechanism (13) to drive the fluorescence sensor (11) to move to the position below a target detection hole position of the sample seat (15), and then controlling the fluorescence sensor (11) to realize fluorescence amplification detection of a sample to be detected;
5) and the main control board (14) controls the fluorescence sensor (11) to sequentially complete the fluorescence amplification detection of all samples to be detected according to the target hole position sequence, and then the detection of the steps 4) to 5) is repeated after the next detection period is reached.
10. The method for realizing high-efficiency constant-temperature PCR (polymerase chain reaction) suitable for the field application of the emergency according to claim 9, wherein the method comprises the following steps:
1) if the hole site detection time in the detection state is up, or the detected abnormal hole site is stopped manually through the start-stop detection (22) of a single detection hole site on the man-machine interaction interface of the screen (2), the hole site is restored to the idle state again;
2) if a new sudden sample to be detected exists and the sample seat is provided with a detection hole site in an idle state, the new sample to be detected is placed into the idle detection hole site, and then the new detection hole site is independently started, so that the hole site in the detection state cannot be influenced, and the high-efficiency constant-temperature PCR device does not need to be interrupted; after the next new detection period comes, adding a target hole position sequence to the current newly started detection hole position, and repeating the detection of the steps 4) -5) by the high-efficiency constant-temperature PCR device;
3) all detection hole sites of same sample seat (15) all can select single detection hole site to open and stop detection (22) to hole site detection time that is in the detection state reachs or can resume idle state again after artificial unusual stopping, as long as there is idle hole site on the sample seat, high-efficient constant temperature PCR device can continuous uninterrupted duty, remains all detection hole sites simultaneously and opens the function that stops the detection, realizes the high-efficient constant temperature amplification of high-efficient constant temperature PCR device through this kind of mode and detects.
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Citations (4)
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- 2019-10-10 CN CN201910957115.2A patent/CN110628607A/en active Pending
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| US20170130261A1 (en) * | 2014-07-08 | 2017-05-11 | National Institute Of Advanced Industrial Science And Technology | Nucleic acid amplification device, nucleic acid amplification method, and chip for nucleic acid amplification |
| CN206768082U (en) * | 2017-04-18 | 2017-12-19 | 朱红 | A kind of magnetic stirring Analysis of Drug Susceptibility instrument and matched reagent box |
| CN107653186A (en) * | 2017-09-15 | 2018-02-02 | 广东达元食品药品安全技术有限公司 | A kind of portable PCR instrument |
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| Title |
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| 单秋霞;李建杰;吴晓;杜与冲;袁旭军;: "恒温PCR双通道荧光读数仪的系统设计", 电子科技, vol. 29, no. 03, 15 March 2016 (2016-03-15), pages 110 - 113 * |
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