Bottom fluorescent quantitative PCR instrument calibration system

Abstract

The utility model relates to the technical field of equipment calibration, in particular to a bottom fluorescent quantitative PCR (Polymerase Chain Reaction) instrument calibration system which is provided with a host unit and a calibration unit which are connected with a computer, so that the calibration of the calibration unit is intelligently completed; the calibration unit is designed into a double-probe structure of the temperature probe unit and the fluorescence output unit, so that the structure is small in size, adapts to temperature optical calibration of the small-flux fluorescence PCR instrument, and is good in flexibility; a temperature sensor of a temperature probe is located at the upper top end, a surface-mounted LED is located at the bottom of an inner cavity of a fluorescence output unit, and a light emitting surface faces upwards, so that the optical detection mode of the small-flux fluorescent quantitative PCR instrument is bottom (including bottom side) fluorescence detection; the temperature probe unit is provided with a temperature probe and a fixed shell which are in threaded connection, and the fluorescence output unit is provided with a color-changing lampshade and a shell which are in threaded connection, so that installation and replacement are convenient; the color-changing lampshade is provided with a plurality of colors corresponding to different fluorescence wavelength ranges, so that the calibration of fluorescence channels of the fluorescence quantitative PCR instrument in different wavelength ranges is realized, the cost is low, and the efficiency is high.

Description

Technical Field

[0001] The utility model relates to the technical field of equipment calibration, in particular to a bottom fluorescence quantitative PCR instrument calibration system. Background Art

[0002] Real-time fluorescence quantitative PCR is a method that uses fluorescent chemicals to monitor the total amount of products after each polymerase chain reaction cycle in a DNA amplification reaction. Real-time fluorescence quantitative PCR monitors the reaction process in real time and collects data. It is simple and fast and is widely used in life science research, clinical disease diagnosis, animal disease diagnosis, food safety testing and other fields. Currently, there are physical devices that can simulate the sample fluorescence amplification process to calibrate the optical temperature parameters of the fluorescence quantitative PCR instrument, but it is mainly suitable for fluorescence quantitative PCR instruments with top fluorescence detection. Although there are a small number of fluorescence PCR instruments suitable for bottom fluorescence detection, they are generally 96-well and 48-well designs with poor flexibility. With the development of PCR instruments, the optical detection mode of some fluorescence quantitative PCR instruments is bottom (including bottom side) fluorescence detection, and the number of small-throughput fluorescence PCR instruments such as 8-well / 16-well / 32-well instruments is increasing. Therefore, a temperature optical calibration system is urgently needed to calibrate them, and different fluorescent dyes are used at different wavelengths, which has high cost and low efficiency. Utility Model Content

[0003] The utility model discloses a bottom fluorescence quantitative PCR instrument calibration system, aiming to solve the technical problems existing in the prior art.

[0004] The utility model adopts the following technical solutions:

[0005] A bottom fluorescence quantitative PCR instrument calibration system is connected to a computer, comprising a host unit and a calibration unit connected to the host unit via a cable;

[0006] The host unit includes a shell and an electronic unit located in the inner cavity of the shell, the electronic unit includes a first circuit board and a controller, a memory, a communication module and an optical drive module located on the first circuit board, and the controller is control-connected to the calibration unit;

[0007] The calibration unit comprises a temperature probe unit and a fluorescence output unit, wherein the temperature probe unit and the fluorescence output unit are installed in the same direction on a base having a second circuit board installed therein;

[0008] The temperature probe unit comprises a detachably connected temperature probe at the upper end and a fixed housing at the lower end, and the temperature probe is provided with a temperature sensing element which is electrically connected to the second circuit board;

[0009] The fluorescent output unit includes a detachably connected color-changing lampshade located at the upper end and an outer shell located at the lower end. The color-changing lampshade is communicated with an inner chamber provided in the outer shell. A surface-mount LED soldered on the second circuit board is provided at the bottom of the inner chamber. The surface-mount LED is located on the central axis of the inner chamber.

[0010] In some embodiments, a power interface, a USB interface and a cable interface are respectively provided on the housing of the host unit, and one end of the cable is connected to the first circuit board, and the other end is connected to the second circuit board.

[0011] In some embodiments, the controller is respectively connected to the optical driving module, the communication module, the memory and the temperature sensing element, and the communication module is connected to the computer for communication.

[0012] In some embodiments, a plane for horizontally fixing the second circuit board is provided at the upper end of the base, and the base is provided with interconnected grooves and end through holes in sequence from top to bottom; the circuit board through hole provided on the second circuit board is located at the upper end of the groove, and the central axes of the circuit board through hole, the groove, and the end through hole all coincide.

[0013] In some embodiments, the fixed shell of the temperature probe unit is provided with an upper column, a lower platform, a boss and a positioning short column formed on the outside of the bottom of the boss, which are connected in sequence from top to bottom. The boss and the positioning short column are respectively embedded in the groove and the end through hole.

[0014] In some embodiments, the upper cylinder is provided with a threaded hole in the upper cylinder, the conical inner cavity in the boss is connected to the threaded hole in the upper cylinder, the small head end of the conical inner cavity is located at the bottom, and the small head end is also provided with a wire entrance for connecting a flexible wire to the second circuit board.

[0015] In some embodiments, the temperature probe includes a probe housing and a temperature sensor located at the top of a central through hole in the probe housing, and the temperature sensor is connected to the second circuit board via a flexible wire; an outer periphery of the lower end of the probe housing is provided with an external probe thread, and the external probe thread is matched and connected with the internal thread of the threaded hole in the upper cylinder; and the central axes of the central through hole, the threaded hole in the upper cylinder, the conical inner cavity and the positioning short column all coincide.

[0016] In some embodiments, the top of the temperature probe and the top of the color-changing lampshade are both gradually tapering cones, and the temperature probe and the color-changing lampshade are respectively matched with two reaction holes provided in the fluorescent quantitative PCR instrument. The parts of the temperature probe and the color-changing lampshade that match the reaction holes have the same size, and the temperature probe fits tightly with the reaction hole, so that the temperature of the reaction hole can be obtained in real time.

[0017] In some embodiments, a blind path is provided inside the color-changing lampshade along the central axis, the blind path opening faces downward, the lower end of the color-changing lampshade is provided with an external thread of the lampshade, the upper end of the outer shell is provided with a shell tube portion, and the lower end is provided with a shell base, the inner wall of the shell tube portion is provided with a shell internal thread, the lampshade external thread is matched and connected with the shell internal thread, the shell base is provided with an inner cavity, a SMD LED is provided at the bottom of the inner cavity with the light-emitting surface facing upward, and the central axis of the SMD LED, the central axis of the blind path, and the central axis of the inner cavity all coincide.

[0018] In some embodiments, the color-changing lampshade has several different colors, each color corresponding to a different fluorescent wavelength range. Beneficial Effects

[0019] The utility model discloses a bottom fluorescence quantitative PCR instrument calibration system. Compared with the prior art, the utility model has the following advantages:

[0020] A bottom fluorescence quantitative PCR instrument calibration system is provided, by setting a host unit and a calibration unit connected to a computer, wherein the host unit is provided with an electronic unit including a controller, a communication module and an optical drive module, and the calibration unit is intelligently completed under the control of the computer; the calibration unit is a temperature probe unit and a fluorescence output unit installed in the same direction, i.e., a double-probe structure, with a small structure volume, suitable for temperature optical calibration of a small-throughput fluorescence PCR instrument, and good flexibility; the top of the temperature probe and the top of the color-changing lampshade are both tapered, the temperature probe and the color-changing lampshade are respectively matched with two reaction holes provided in the fluorescence quantitative PCR instrument, and the temperature The temperature sensor of the probe is located at the top, and the SMD LED is located at the bottom of the inner cavity of the fluorescence output unit with the light-emitting surface facing upward, so that the optical detection mode of the small-throughput fluorescence quantitative PCR instrument can be realized as bottom (including bottom side) fluorescence detection. The temperature probe unit is provided with a temperature probe at the upper end and a fixed shell at the lower end which are threadedly connected, and the fluorescence output unit includes a color-changing lampshade at the upper end and an outer shell at the lower end which are threadedly connected, so that installation and replacement are convenient. The color-changing lampshade is provided with several different colors, and each color corresponds to a different fluorescence wavelength range, so that the fluorescence channels of the fluorescence quantitative PCR instrument with different wavelength ranges can be calibrated, and the use cost is low and the efficiency is high. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the technical solution of the embodiment of the utility model, the following is a brief introduction to the drawings required for the description of the embodiment, which constitute a part of the utility model. The schematic embodiments of the utility model and their descriptions explain the utility model and do not constitute an improper limitation on the utility model; in the drawings:

[0022] Figure 1 It is a schematic diagram of the technical solution structure of the bottom fluorescence quantitative PCR instrument calibration system disclosed in the embodiment of the utility model;

[0023] Figure 2 A schematic diagram of the technical solution structure of the calibration unit disclosed in the embodiment of the utility model;

[0024] Figure 3 for Figure 1 A-direction view.

[0025] In the figure:

[0026] Host unit 1; housing 11; power interface 111; USB interface 112; cable interface 113; first circuit board 12; controller 121; communication module 122; memory 123; optical drive module 124; calibration unit 2; base 21; groove 211; end through hole 212; temperature probe unit 22; temperature probe 221; probe housing 2211; temperature sensing element 2212; flexible wire 2213; center through hole 2214; probe external thread 2215; fixed housing 222; threaded hole 2221 in upper column; upper column Body 2222; lower body 2223; boss 2224; positioning short column 2225; wire inlet 2226; tapered inner cavity 2227; fluorescent output unit 23; color-changing lampshade 231; channel 2311; lampshade external thread 2312; outer shell 232; shell tube 2321; shell internal thread 2322; shell base 2323; inner cavity 2324; SMD LED 233: second circuit board 24; circuit board through hole 241; flat cable 3; fastening screw 4; computer 5. DETAILED DESCRIPTION

[0027] In order to make the purpose, technical solution and advantages of the utility model clearer, the technical solution of the utility model will be clearly and completely described below in combination with the specific embodiments of the utility model and the corresponding drawings; in the description of the utility model, it should be noted that the "including" mentioned in the specification and the claims is an open term, so it should be interpreted as "including but not limited to"; "several" refers to more than 3.

[0028] In the description of the present invention, it should be noted that the terms "center", "up", "down", "left", "right", "vertical", "horizontal", "inside", "outside", etc. indicate directions or positional relationships based on the directions or positional relationships shown in the accompanying drawings. They are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific direction, be constructed and operated in a specific direction. Therefore, they cannot be understood as limitations on the present invention.

[0029] Obviously, the described embodiments are only part of the embodiments of the utility model, rather than all of the embodiments; based on the embodiments in the utility model, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the utility model.

[0030] like Figure 1-Figure 3 As shown, the technical solution disclosed by the utility model is:

[0031] A bottom fluorescence quantitative PCR instrument calibration system is connected to a computer 5 and comprises a host unit 1 and a calibration unit 2 connected to the host unit 1 via a cable 32 .

[0032] The host unit 1 includes a shell 11 and an electronic unit located in the inner cavity of the shell 11, the electronic unit includes a first circuit board 12 and a controller 121, a memory 123, a communication module 122 and an optical drive module 124 located on the first circuit board 12, and the controller 121 is controlled and connected to the calibration unit 2.

[0033] The calibration unit 2 includes a temperature probe unit 22 and a fluorescence output unit 23, which are installed in the same direction and vertically on a base 21 with a second circuit board 24 installed therein, and has a dual-probe structure.

[0034] The temperature probe unit 22 includes a detachably connected temperature probe 221 at the upper end and a fixed housing 222 at the lower end. The temperature probe 221 is provided with a temperature sensing element 2212 which is electrically connected to the second circuit board 24 .

[0035] The fluorescent output unit 23 includes a detachably connected color-changing lampshade 231 located at the upper end and an outer shell 232 located at the lower end. The color-changing lampshade 231 is communicated with an inner chamber provided in the outer shell 232. A SMD LED 233 soldered on the second circuit board 24 is provided at the bottom of the inner chamber. The SMD LED 233 is located on the central axis of the inner chamber.

[0036] like Figure 1 to Figure 3 As shown, the preferred embodiment provided by the utility model is:

[0037] A bottom fluorescence quantitative PCR instrument calibration system is connected to a computer 5 and comprises a host unit 1 and a calibration unit 2 connected to the host unit 1 via a cable 32 .

[0038] The host unit 1 includes a shell 11 and an electronic unit located in the inner cavity of the shell 11, the electronic unit includes a first circuit board 12 and a controller 121, a memory 123, a communication module 122 and an optical drive module 124 located on the first circuit board 12, and the controller 121 is controlled and connected to the calibration unit 2.

[0039] The housing 11 of the host unit 1 is provided with a power interface 111, a USB interface 112 and a cable interface 112, one end of the cable 3 is connected to the first circuit board 12, and the other end is connected to the second circuit board 24; the power interface 111 is a Type-C interface, which is used to connect a power source to power the entire system, and the USB interface 112 is used to connect to the computer 5, establish a connection with the computer 5 software, and control the system.

[0040] The controller 121 is connected to the computer 5 , and the controller 121 controls the temperature probe 221 to collect the temperature in the reaction hole of the fluorescence quantitative PCR instrument. The temperature data is then transmitted to the controller 121 , stored in the memory 123 , and transmitted to the computer 5 by the communication module 122 .

[0041] The controller 121 controls the optical driving module 124 to work, and the controller 121 controls the communication module 122 and the memory 123 .

[0042] The communication module 122 includes a Hall module, a crystal oscillator and a Bluetooth communication device, and can be connected to the computer 5 for communication to transmit the temperature data information stored in the memory 123 .

[0043] The calibration unit 2 includes a temperature probe unit 22 and a fluorescence output unit 23, which are installed on the base 21 in the same direction and vertically. The temperature probe unit 22 and the fluorescence output unit 23 are respectively matched and installed with two reaction holes (not shown in the figure) arranged side by side in the fluorescence quantitative PCR instrument, and are used for the fluorescence detection of the bottom (including the bottom side) of the optical detection mode of the fluorescence quantitative PCR instrument. The temperature probe 221 and the color-changing lampshade 231 have the same size as the matching parts of the reaction holes. The dual-probe structure has a small volume and is suitable for the temperature optical calibration of small-throughput fluorescence PCR instruments such as 8-hole / 16-hole / 32-hole, and has good flexibility.

[0044] The upper end of the base 21 is provided with a plane for horizontally fixing the second circuit board 24, and the base 21 is provided with interconnected grooves 211 and end through holes 212 in sequence from top to bottom; the circuit board through hole 241 provided on the second circuit board 24 is located at the upper end of the groove 211, and the central axes of the circuit board through hole 241, the groove 211, and the end through hole 212 all coincide.

[0045] The temperature probe unit 22 includes a detachably connected temperature probe 221 at the upper end and a fixed housing 222 at the lower end. The temperature probe 221 is provided with a temperature sensing element 2212 which is electrically connected to the second circuit board 24 .

[0046] The fixed shell 222 of the temperature probe unit 22 is provided with an upper column 2222, a lower platform 2223, a boss 2224 and a positioning short column 2225 formed at the bottom of the boss 2224, which are connected in sequence from top to bottom. The boss 2224 and the positioning short column 2225 are respectively embedded in the groove 211 and the end through hole 212. The fixed shell 222 is made of PEEK material.

[0047] The upper column 2222 is provided with a threaded hole 2221 in the upper column, and the conical inner cavity 2227 provided in the boss 2224 is connected to the threaded hole 2221 in the upper column. The small head end of the conical inner cavity 2227 is located at the bottom, and the small head end is also provided with a wire entry 2226 for connecting the flexible wire 2213 with the second circuit board 24.

[0048] The temperature probe 221 includes a probe housing 2211 and a temperature sensor 2212 located at the top of a central through hole 2214 in the probe housing 2211. In this example, the probe housing 2211 is made of copper, the temperature sensor 2212 is a thermistor, and the temperature sensor 2212 is connected to the second circuit board 24 via a flexible wire 2213; an outer probe thread 2215 is provided on the periphery of the lower end of the probe housing 2211, and the outer probe thread 2215 is matched and connected with the inner thread of the threaded hole 2221 in the upper column; and the central axes of the central through hole 2214, the threaded hole 2221 in the upper column, the conical inner cavity 2227 and the positioning short column 2225 all coincide.

[0049] The top of the temperature probe 221 and the top of the color-changing lampshade 231 are both gradually tapering cones. The temperature probe 221 and the color-changing lampshade 231 are respectively matched with two reaction holes provided in the fluorescent quantitative PCR instrument, and the temperature probe 221 fits tightly with the temperature measuring reaction hole, so that the temperature of the temperature measuring reaction hole can be obtained in real time by the temperature probe 221.

[0050] The fluorescent output unit 23 includes a detachably connected color-changing lampshade 231 located at the upper end and an outer shell 232 located at the lower end. In this example, the color-changing lampshade 231 is set to be a colored PE material, and the outer shell 232 is set to be a PEEK material; the color-changing lampshade 231 is communicated with an inner chamber provided with the outer shell 232, and a chip LED 233 is provided at the bottom of the inner chamber and is soldered on the second circuit board 24, and the chip LED 233 is located on the central axis of the inner chamber.

[0051] A blind path 2311 is provided inside the color-changing lampshade 231 along the central axis, the blind path 2311 opens downward, and the blind end reaches the top of the color-changing lampshade 231. The lower end of the color-changing lampshade 231 is provided with a lampshade external thread 2312. The upper end of the outer shell 232 is provided with a shell tube 2321, and the lower end is provided with a shell pedestal 2323, and the shell pedestal 2323 is fixed to the base 21 by fastening screws 4; the inner wall of the shell tube 2321 is provided with a shell internal thread 2322, the lampshade external thread 2312 is matched and connected with the shell internal thread 2322, and the color-changing lampshade 231 can be screwed onto the outer shell 232 and fixed, and the shell pedestal 2323 is provided with an inner cavity 2324, and a patch LED 233 is provided at the bottom of the inner cavity 2324 with the light-emitting surface facing upward, and the central axis of the patch LED 233, the central axis of the blind path 2311, and the central axis of the inner cavity 2324 all coincide.

[0052] The color-changing lampshade 231 is provided with several colors, each color corresponds to a different fluorescence wavelength range. By replacing the color-changing lampshades 231 of different colors, the fluorescence channels of different wavelength ranges of the fluorescence quantitative PCR instrument can be calibrated. In this example, the color-changing lampshade has 4 colors to choose from, which can respectively transmit 520-530nm, 560-580nm, 620-650nm, and 680-730nm light. According to the fluorescence channel to be calibrated, the user can rotate the corresponding color-changing lampshade 231 to the position of the SMD LED 233 for calibration. It is easy to use and can be replaced freely; the light emitted by the SMD LED 233 can be evenly transmitted through the color-changing lampshade 231 to collect the changes in the optical signal of the calibrated fluorescence PCR instrument.

[0053] The optical change control of the SMD LED 233 is connected to the optical driving module 124 by the controller 121. The optical driving module 124 controls the current supplied to the SMD LED 233 so that it can change according to the standard S curve according to the change of the fluorescence value during gene amplification, and is collected as an output signal by the calibrated fluorescence PCR instrument.

[0054] The optical change control signal of the SMD LED 233 comes from the temperature probe 221. When the temperature probe 221 collects a set of high and low temperature changes, it is transmitted to the controller 121. The controller 121 drives the optical driving module 124 to give the SMD LED 233 a change in current. The increase in current increases the luminous intensity of the SMD LED 233, simulating the sample fluorescence change in the actual amplification process. The fluorescent PCR instrument collects the optical signal changes, and the obtained data is used for calculation to give a calibration result.

[0055] The embodiments of the utility model are described above in conjunction with the accompanying drawings, but the utility model is not limited to the above-mentioned specific implementation methods. The above-mentioned specific implementation methods are merely illustrative and not restrictive. Under the enlightenment of the utility model, ordinary technicians in this field can also make many forms without departing from the scope of protection of the utility model and the claims, all of which are within the protection of the utility model.

Claims

1. A bottom fluorescence quantitative PCR instrument calibration system, connected to a computer, characterized in that: It includes a host unit and a calibration unit connected to the host unit via a cable; The host unit includes a shell and an electronic unit located in the inner cavity of the shell, the electronic unit includes a first circuit board and a controller, a memory, a communication module and an optical drive module located on the first circuit board, and the controller is control-connected to the calibration unit; The calibration unit comprises a temperature probe unit and a fluorescence output unit, wherein the temperature probe unit and the fluorescence output unit are installed in the same direction on a base having a second circuit board installed therein; The temperature probe unit comprises a detachably connected temperature probe at the upper end and a fixed housing at the lower end, and the temperature probe is provided with a temperature sensing element which is electrically connected to the second circuit board; The fluorescent output unit includes a detachably connected color-changing lampshade located at the upper end and an outer shell located at the lower end. The color-changing lampshade is communicated with an inner chamber provided in the outer shell. A surface-mount LED soldered on the second circuit board is provided at the bottom of the inner chamber. The surface-mount LED is located on the central axis of the inner chamber.

2. The bottom fluorescence quantitative PCR instrument calibration system according to claim 1, characterized in that: The housing of the host unit is provided with a power interface, a USB interface and a cable interface respectively. One end of the cable is connected to the first circuit board, and the other end is connected to the second circuit board.

3. The bottom fluorescence quantitative PCR instrument calibration system according to claim 2, characterized in that: The controller is respectively connected to the optical driving module, the communication module, the memory and the temperature sensing element, and the communication module is connected to the computer for communication.

4. The bottom fluorescence quantitative PCR instrument calibration system according to claim 1, characterized in that: The upper end of the base is provided with a plane for horizontally fixing the second circuit board, and the base is provided with interconnected grooves and end through holes in sequence from top to bottom; the circuit board through hole provided on the second circuit board is located at the upper end of the groove, and the central axes of the circuit board through hole, the groove, and the end through hole all coincide.

5. The bottom fluorescence quantitative PCR instrument calibration system according to claim 4, characterized in that: The fixed shell of the temperature probe unit is provided with an upper column, a lower table, a boss and a positioning short column formed at the bottom of the boss in sequence from top to bottom. The boss and the positioning short column are respectively embedded in the groove and the end through hole.

6. The bottom fluorescence quantitative PCR instrument calibration system according to claim 5, characterized in that: The upper cylinder is provided with an internal threaded hole, the boss is provided with a conical inner cavity which is connected with the internal threaded hole, the small end of the conical inner cavity is located at the bottom, and the small end is also provided with a wire inlet for connecting a flexible wire to the second circuit board.

7. The bottom fluorescence quantitative PCR instrument calibration system according to claim 6, characterized in that: The temperature probe includes a probe housing and a temperature sensor located at the top of a central through hole in the probe housing, and the temperature sensor is connected to the second circuit board via a flexible wire; an outer periphery of the lower end of the probe housing is provided with a probe external thread, and the probe external thread is matched and connected with the internal thread of the threaded hole in the upper cylinder; and the central axes of the central through hole, the threaded hole in the upper cylinder, the tapered inner cavity and the positioning short column all coincide.

8. The bottom fluorescence quantitative PCR instrument calibration system according to claim 5, characterized in that: The top of the temperature probe and the top of the color-changing lampshade are both gradually decreasing cones. The temperature probe and the color-changing lampshade are respectively matched with two reaction holes provided in the fluorescent quantitative PCR instrument. The parts of the temperature probe and the color-changing lampshade that match the reaction holes have the same size, and the temperature probe fits tightly with the reaction hole, so that the temperature of the reaction hole can be obtained in real time.

9. The bottom fluorescence quantitative PCR instrument calibration system according to claim 1, characterized in that: A blind path is provided inside the color-changing lampshade along the central axis, and the opening of the blind path faces downward. The lower end of the color-changing lampshade is provided with an external thread of the lampshade, the upper end of the outer shell is provided with a shell tube portion, and the lower end is provided with a shell base. The inner wall of the shell tube portion is provided with an internal shell thread, and the external thread of the lampshade is matched and connected with the internal thread of the shell. The shell base is provided with an inner cavity, and a SMD LED is provided at the bottom of the inner cavity with the light-emitting surface facing upward. The central axis of the SMD LED, the central axis of the blind path, and the central axis of the inner cavity all coincide.

10. The bottom fluorescence quantitative PCR instrument calibration system according to claim 9, characterized in that: The color-changing lampshade is provided with a plurality of different colors, and each color corresponds to a different fluorescent wavelength range.

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