Optical detection device and PCR analyzer

Abstract

The utility model relates to an optical detection device and a PCR analyzer. The PCR analyzer comprises a temperature control amplification device and an optical detection device which are connected with each other through optical fibers, the optical detection device comprises an optical fiber fixing seat, a rotating disc and a plurality of detection head assemblies, the optical fiber fixing seat is provided with a plurality of optical fiber fixing holes which are arranged around a central axis at intervals, and each optical fiber fixing hole is used for insertion of an optical fiber; the rotating disc is rotatably connected to the optical fiber fixing seat, the plurality of detection head assemblies are arranged on the rotating disc around the central axis, and the distance between any two adjacent detection head assemblies is greater than or equal to the distance between two adjacent optical fiber fixing holes, so that the rotating disc rotates for one circle; the sample of each PCR tube in the sample carrier can be irradiated by exciting light with different wavelengths, so that positioning errors caused by line feed and column feed in an X-Y direction scanning mode do not exist, the samples can be comprehensively analyzed in a short time, the waiting time can be saved, and the detection and analysis efficiency can be improved.

Description

Technical Field

[0001] This application relates to the field of biological detection technologies, and particularly to an optical detection device and a PCR analyzer. Background Art

[0002] Polymerase chain reaction (PCR) is a molecular biology technique used to amplify specific DNA fragments. It can be regarded as a special DNA amplification outside the organism. This technique can greatly increase trace gene fragments. A PCR analyzer (gene amplifier) is a gene amplification instrument that uses PCR technology for gene analysis applications.

[0003] When a PCR analyzer is running, real-time fluorescence quantitative PCR technology needs to be used, that is, a fluorescent group is added to the PCR reaction system, and the entire PCR process is detected in real time through the change of the fluorescent signal during the PCR reaction. Finally, quantitative analysis of the sample to be tested is performed through a standard curve. Therefore, a corresponding optical detection device is needed to ensure the stable state of the excitation light and fluorescence. Currently, most existing optical detection devices use a scanning method in the X-Y direction for detection. This method has a slow detection speed, not only consumes a large amount of time, but also has positioning errors when changing rows and columns. It seriously affects the stability of the channel signal and also limits the shortening of the working time. Summary of the Utility Model

[0004] Based on this, the purpose of this application is to provide an optical detection device and a PCR analyzer including the optical detection device to solve the problems that the optical detection device of the existing PCR analyzer has a slow detection speed and there are positioning errors when changing rows and columns.

[0005] According to one aspect of this application, an optical detection device is provided, including:

[0006] An optical fiber fixing seat, the optical fiber fixing seat is provided with a plurality of optical fiber fixing holes arranged at intervals around a central axis, and each of the optical fiber fixing holes is used for inserting an optical fiber;

[0007] A rotating disk, rotatably connected to the optical fiber fixing seat, and the rotating disk can rotate around the central axis;

[0008] A plurality of detection head components, arranged on the rotating disk around the central axis, and the distance between any two adjacent detection head components is greater than or equal to the distance between two adjacent optical fiber fixing holes; the detection head components can follow the rotation of the rotating disk so that each detection head component can sequentially face each optical fiber fixing hole.

[0009] In one embodiment, the distance between two adjacent ones of the optical fiber fixing holes is greater than or equal to the aperture of the optical fiber fixing holes.

[0010] In one embodiment, the detection head assembly includes a housing, a light source, a first filter, a dichroic mirror, a second filter, and a detector, wherein the light source, the first filter, the dichroic mirror, the second filter, and the detector are disposed in the housing, and the light source, the first filter, and the dichroic mirror are arranged at intervals along a first straight line; the dichroic mirror, the second filter, and the detector are arranged at intervals along a second straight line perpendicular to the first straight line, and the dichroic mirror is inclined at 45° relative to the first straight line or the second straight line so that the light emitted by the light source can be refracted in a direction away from the detector.

[0011] In one embodiment, a focusing lens is further disposed in the housing, and the focusing lens is disposed on a side of the dichroic mirror away from the second filter on the second straight line.

[0012] In one embodiment, at least part of the material of the housing is an opaque material, and / or the housing is grounded.

[0013] In one embodiment, the optical detection device includes a conductive slip ring and a cable, the conductive slip ring is coaxially mounted on the rotating disk, one end of the cable is connected to the detection head assembly, and the other end is connected to the conductive slip ring.

[0014] In one embodiment, a plurality of rolling balls are mounted on a side of the rotating disk facing the optical fiber fixing seat in a rolling manner, and the rolling balls are in rolling connection with the optical fiber fixing seat.

[0015] In one embodiment, a driving element drivingly connected to the rotating disk is mounted on the optical fiber fixing seat, and the driving element is used to drive the rotating disk to rotate; a sensing element is mounted on one of the optical fiber fixing seat and the rotating disk, and a limiting piece is mounted on the other, and the sensing element is configured to monitor the rotation state of the driving element driving the rotating disk when being aligned with the limiting piece.

[0016] In one embodiment, a plurality of mounting seats are mounted on the edge of the rotating disk, the plurality of mounting seats are arranged at intervals around the central axis, and each detection head assembly is fixedly mounted on a corresponding one of the mounting seats.

[0017] According to another aspect of the present application, there is provided a PCR analyzer, including a temperature control amplification device and the optical detection device as described in any one of the above, and the optical detection device is connected to the temperature control amplification device through an optical fiber.

[0018] In the above optical detection device and PCR analyzer, a plurality of optical fiber fixing holes around a central axis are provided on the optical fiber fixing base, and a plurality of detection head assemblies around the central axis are arranged on the rotating disk. The distance between any two adjacent detection head assemblies is equal to the distance between two adjacent optical fiber fixing holes. When the rotating disk rotates one circle, the samples to be tested in each PCR tube in the sample carrier can be sequentially irradiated by excitation lights of different wavelengths. Therefore, compared with the optical detection device of the existing PCR analyzer that adopts the X-Y direction scanning method, there is no positioning error caused by line and column changing in the X-Y direction scanning method, and the samples to be tested can be comprehensively analyzed in a short time (such as 1 second to 2 seconds). Therefore, the waiting time can be saved and the detection and analysis efficiency can be improved. Description of the Drawings

[0019] Figure 1 Isometric view of the optical detection device provided by an embodiment of the present application Figure One 。

[0020] Figure 2 Isometric view of the optical detection device provided by an embodiment of the present application Figure Two 。

[0021] Figure 3 Cross-sectional view of the optical detection device provided by an embodiment of the present application.

[0022] Figure 4 Isometric view of the sample carrier provided by an embodiment of the present application.

[0023] Figure 5 Exploded view of the detection head assembly in the optical detection device provided by an embodiment of the present application.

[0024] Description of the Reference Numerals:

[0025] 10. Optical detection device; 100. Optical fiber fixing base; 101. Optical fiber fixing hole; 200. Driving element; 300. Rotating disk; 400. Detection head assembly; 410. Housing; 420. Light source; 430. First filter; 440. Dichroic mirror; 450. Second filter; 460. Detector; 470. Focusing lens; 500. Scanning control board; 600. Rolling ball; 700. Conductive slip ring; 800. Sensing element; 900. Limiting piece; 20. Central axis; 30. Sample carrier; 31. PCR tube; 40. First straight line; 50. Second straight line. Detailed Embodiments

[0026] To make the above objects, features, and advantages of the present application more apparent and understandable, the following provides a detailed description of the specific embodiments of the present application in conjunction with the accompanying drawings. A lot of specific details are set forth in the following description to facilitate a full understanding of the present application. However, the present application can be implemented in many other ways different from those described herein. Those skilled in the art can make similar improvements without departing from the connotation of the present application. Therefore, the present application is not limited by the specific embodiments disclosed below.

[0027] In the description of the present application, it should be understood that if terms such as "center", "longitudinal", "transverse", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", "axial", "radial", "circumferential", etc. appear, the orientation or positional relationship indicated by these terms is based on the orientation or positional relationship shown in the drawings, and is only for the convenience of describing the present application and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be construed as a limitation to the present application.

[0028] In addition, if terms such as "first" and "second" appear, these terms are only for descriptive purposes and should not be construed as indicating or implying relative importance or implicitly specifying the quantity of the indicated technical features. Thus, features defined with "first" and "second" may explicitly or implicitly include at least one such feature. In the description of the present application, if the term "plurality" appears, the meaning of "plurality" is at least two, such as two, three, etc., unless otherwise specifically and clearly defined.

[0029] In the present application, unless otherwise clearly specified and limited, if terms such as "install", "connect", "join", "fix", etc. appear, these terms should be understood in a broad sense. For example, it may be a fixed connection, a detachable connection, or integrated; it may be a mechanical connection or an electrical connection; it may be directly connected or indirectly connected through an intermediate medium, and it may be the communication inside two elements or the interaction relationship between two elements, unless otherwise clearly limited. For those of ordinary skill in the art, the specific meanings of the above terms in the present application can be understood according to specific circumstances.

[0030] In this application, unless otherwise clearly defined and limited, if there is a description such as a first feature being "on" or "under" a second feature, its meaning can be that the first and second features are in direct contact, or the first and second features are in indirect contact through an intermediate medium. Moreover, the first feature being "above", "over" and "on top of" the second feature can mean that the first feature is directly above or obliquely above the second feature, or simply indicates that the first feature has a higher horizontal height than the second feature. The first feature being "under", "beneath" and "underneath" the second feature can be that the first feature is directly below or obliquely below the second feature, or simply indicates that the first feature has a lower horizontal height than the second feature.

[0031] It should be noted that if an element is referred to as being "fixed to" or "disposed on" another element, it can be directly on the other element or there can also be an intermediate element. If an element is considered to be "connected" to another element, it can be directly connected to the other element or there may be an intermediate element at the same time. If so, the terms "vertical", "horizontal", "upper", "lower", "left", "right" and similar expressions used in this application are only for the purpose of illustration and do not represent the only implementation.

[0032] This application provides an optical detection device and a PCR analyzer. The PCR analyzer includes the optical detection device. The PCR analyzer is used to simulate the DNA replication process in vivo in vitro based on the amplification principle of polymerase chain reaction (PCR). The optical detection device is used to perform real-time detection on the entire PCR process based on real-time fluorescence quantitative PCR technology, and finally perform quantitative analysis on the sample to be tested through a standard curve.

[0033] The structures of the PCR analyzer and the optical detection device in this application will be described below. It can be understood that in other embodiments, the optical detection device of this application is not limited to being only used in the PCR analyzer, and can also be used in any device that requires optical detection of samples, which is not limited here.

[0034] The PCR analyzer provided by the embodiment of this application includes a temperature control amplification device (not shown in the figure) and the Figures 1 to 3 shown optical detection device 10. The optical detection device 10 is connected to the temperature control amplification device through an optical fiber (not shown in the figure). The temperature control amplification device is used to perform cycles of high-temperature denaturation, low-temperature annealing and appropriate-temperature extension on the sample to be tested to achieve rapid amplification of the target DNA fragment; the optical detection device 10 is used to emit excitation light to the sample to be tested and receive the fluorescence emitted by the sample to be tested, so as to facilitate subsequent analysis of the sample to be tested, so as to be able to monitor the reaction process of PCR in real time.

[0035] Specifically, in one embodiment, as Figure 1 and Figure 2As shown, the optical detection device 10 includes an optical fiber fixing base 100, a driving element 200, a rotating disk 300, and a plurality of detection head assemblies 400. The optical fiber fixing base 100 is provided with a plurality of optical fiber fixing holes 101 arranged at intervals around a central axis 20. Each optical fiber fixing hole 101 is correspondingly inserted with an optical fiber. One end of the optical fiber is connected to a temperature-controlled amplification device, and the end far from the temperature-controlled amplification device is inserted into the optical fiber fixing hole 101. The reason for setting a plurality of optical fiber fixing holes 101 is as Figure 4 shown. The sample carrier 30 for holding the sample to be tested has a plurality of PCR tubes 31. Therefore, a plurality of optical fiber fixing holes 101 are also provided, and the number of optical fiber fixing holes 101 is equal to the number of PCR tubes 31 in the sample carrier 30, so that each optical fiber can transmit an optical signal to a corresponding PCR tube 31 and can transmit the fluorescence signal emitted from the sample to be tested in the corresponding PCR tube 31.

[0036] The rotating disk 300 is rotatably connected to the optical fiber fixing base 100. A plurality of detection head assemblies 400 are arranged on the rotating disk 300 around the central axis 20. The distance between any two adjacent detection head assemblies 400 is greater than or equal to the distance between two adjacent optical fiber fixing holes 101 to prevent mutual crosstalk between adjacent optical fibers. The driving element 200 is fixedly installed on the optical fiber fixing base 100 and is connected to the rotating disk 300. Under the drive of the driving element 200, the rotating disk 300 rotates around the central axis 20 (i.e., around its own central axis 20), so that each optical fiber fixing hole 101 can sequentially face each detection head assembly 400, so that the sample to be tested in each accommodation cavity can be sequentially irradiated by the excitation light emitted by each detection head assembly 400, and each detection head assembly 400 can receive the fluorescence emitted by the sample to be tested. Preferably, the distance between two adjacent optical fiber fixing holes 101 is greater than or equal to the core diameter of the optical fiber fixing hole 101 to avoid signal mutual crosstalk.

[0037] Exemplarily, in Figure 2 the embodiment shown, the number of detection head assemblies 400 is eight. Therefore, it means that the optical detection device 10 has eight channels and can emit eight different wavelengths of excitation light, so the high-throughput performance of the experiment can be improved. It can be understood that the number of detection head assemblies 400 is not limited and is not specifically defined.

[0038] More specifically, in Figure 2In the embodiment shown, a scanning control board 500 for controlling the detection head assembly 400 is provided on the rotating disk 300. The detection head assembly 400 is connected to the scanning control board 500 and is connected to the rotating disk 300 through the scanning control board 500. To further prevent the detection head assembly 400 from falling off the rotating disk 300, more preferably, a plurality of mounting seats are installed on the edge of the rotating disk 300. The plurality of mounting seats are arranged at intervals around the central axis 20, and each detection head assembly 400 is fixedly installed on a corresponding mounting seat, so that the detection head assembly 400 can be more firmly fixedly installed on the rotating disk 300.

[0039] As described above, the optical detection device 10 performs real-time detection on the entire PCR process based on the real-time fluorescence quantitative PCR technology. The specific analysis principle is to add a fluorescent group in the amplification reaction, use the accumulation of fluorescent signals to monitor the entire PCR process in real time, and finally perform quantitative analysis on the test sample through a standard curve. Therefore, each detection head assembly 400 can generate excitation light, which is transmitted to the test samples in each PCR tube 31 of the sample carrier 30 through an optical fiber. If the known fragments for characterizing various diseases in the test sample match the wavelength of the excitation light, the fluorescent group in the test sample will be excited to generate a fluorescent signal. The detection circuit of the detection head assembly 400 performs real-time detection on the test samples in each PCR tube 31 of the sample carrier 30. At the same time, the fluorescent signal is plotted as a curve as the number of cycles changes, and finally the test sample is quantitatively analyzed through a standard curve.

[0040] In this way, as long as the rotating disk 300 rotates one circle, the test samples in each PCR tube 31 of the sample carrier 30 can be irradiated by different wavelengths of excitation light in turn, so that the test samples can be comprehensively analyzed in a short time (for example, 1 second to 2 seconds). Therefore, the waiting time can be saved and the detection and analysis efficiency can be improved.

[0041] Furthermore, as Figure 3 shown, a plurality of rolling balls 600 are installed on the side of the rotating disk 300 facing the optical fiber fixing seat 100 in a rolling manner. All the rolling balls 600 are arranged at intervals around the central axis 20 of the rotating disk 300, and the rolling balls 600 are in rolling connection with the optical fiber fixing seat 100. In this way, it can be ensured that the rotating disk 300 remains stable during rotation and avoids yaw.

[0042] Even further, as Figure 2 and Figure 3As shown. The optical detection device 10 further includes a conductive slip ring 700 and a cable (not shown in the figure). The conductive slip ring 700 is coaxially mounted on the rotating disk 300. One end of the cable is connected to the detection head assembly 400, and the other end is connected to the conductive slip ring 700, so that an external control device can supply power to the optical detection device 10 and perform signal processing. And the conductive slip ring 700 can rotate together with the rotating disk 300, so that when the rotating disk 300 rotates, the cable will not rotate together with the rotating disk 300, so that they will not be wound around each other, thereby ensuring that the rotating disk 300 and the detection head assembly 400 can rotate continuously by 360°, so that continuous detection can be performed in multiple cycles.

[0043] Optionally, as Figure 2 shown, the optical fiber fixing seat 100 is installed with a sensing element 800 (not shown in the figure), and the rotating disk 300 is installed with a limiting piece 900. The sensing element 800 is configured to monitor the rotation state of the driving element 200 driving the rotating disk 300 when it is aligned with the limiting piece 900. For example, monitor the positions where the driving element 200 starts and stops rotating the rotating disk 300, and monitor the positions where the rotating disk 300 starts and stops rotating at a constant speed, so as to realize the positioning of the rotating disk 300. Of course, it can also be that the sensing element 800 is installed on the rotating disk 300 and the limiting piece 900 is installed on the optical fiber fixing seat 100, which is not limited here.

[0044] In the internal structure of the detection head assembly 400, as Figure 5 shown, each detection head assembly 400 includes a housing 410, a light source 420, a first filter 430, a dichroic mirror 440, a second filter 450 and a detector 460. Among them, the light source 420, the first filter 430, the dichroic mirror 440, the second filter 450 and the detector 460 are arranged in the housing 410, and the light source 420, the first filter 430, the dichroic mirror 440 are arranged at intervals along a first straight line; the dichroic mirror 440, the second filter 450 and the detector 460 are arranged at intervals along a second straight line perpendicular to the first straight line. The dichroic mirror 440 is placed at a 45° angle, so that the light emitted by the light source 420 can be refracted in a direction away from the detector 460.

[0045] Preferably, a focusing lens 470 is further provided in the housing 410. The focusing lens 470 is arranged on the second straight line on the side of the dichroic mirror 440 away from the second filter 450. It is used to converge multiple parallel light rays into a concentrated beam when the light irradiates, so that most of the light emitted by the light source 420 can enter the optical fiber for transmission to avoid light loss.

[0046] In a preferred embodiment, the light source 420 uses a collimated and channel-monochromatic LED lamp, such that the light emitted by the LED lamp is not diffused light but straight light. Therefore, there is no need to add a lens at the position of the light source 420, thus simplifying the system design. Moreover, the use of a monochromatic LED lamp can avoid the spectral overlap problem that may be caused by a multi-color light source, ensuring the specificity and accuracy of detection. Using a monochromatic LED can precisely control the wavelength of the excitation light, thereby only exciting specific fluorescent groups and avoiding interference from light of other wavelengths to the signal, improving the sensitivity and accuracy of detection. In addition, compared with traditional light sources such as mercury lamps, the monochromatic LED lamp has a longer service life and lower energy consumption, and is more suitable for PCR experiments that require long-term operation.

[0047] During detection, the excitation light emitted by the light source 420 is filtered by the first filter 430 to remove stray light. After being refracted by 90° on the dichroic mirror 440, it is focused by the focusing lens 470 and then enters the optical fiber. The optical fiber conducts it to the sample carrier 30 and into the solution of the sample to be tested contained in the corresponding PCR tube 31. If the wavelength of the excitation light matches the known fragment of the sample to be tested, the solution of the sample to be tested will be excited to generate fluorescence. The optical fiber conducts the fluorescence back to the focusing lens 470, and then the focusing lens 470 collects and collimates it and incident on the dichroic mirror 440. After passing through the dichroic mirror 440, it passes through the second filter 450 to filter out stray light, and finally enters the detector 460 for detection of the fluorescence signal. The signals do not interfere with each other, and it can save costs and improve space utilization. By using one optical fiber, the sample to be tested can be sequentially irradiated by multiple (for example, eight) different wavelengths of excitation light. By using multiple optical fibers, the samples to be tested in all PCR tubes 31 can be detected simultaneously, improving the number of PCR detections and the detection efficiency.

[0048] In addition, in some better embodiments, at least part of the material of the housing 410 is an opaque material (such as black silicone or surface-treated matte black). Therefore, it can reduce the influence of the external environment on light, and the housing 410 can also be grounded to weaken the signal interference of 50 Hz of the earth, so as to provide stable and reliable signal transmission and ensure the accuracy and stability of the PCR detection result.

[0049] It can be seen that compared with the X-Y direction scanning method used in existing PCR analyzers, the above optical detection device 10 does not have the positioning error existing in the X-Y direction scanning method due to line and column changes. Moreover, in the case where it usually takes 8 seconds to 10 seconds to complete six-channel detection, the optical detection device 10 provided by the present application has a faster scanning speed and only takes 1 second to 2 seconds to complete eight-channel detection. Therefore, the detection efficiency is greatly improved.

[0050] The technical features of the above-described embodiments can be combined arbitrarily. For the sake of brevity of description, not all possible combinations of the various technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, it should be considered to be within the scope described in this specification.

[0051] The above-described embodiments merely represent several implementation manners of the present application. The description is relatively specific and detailed, but it should not be construed as a limitation on the scope of the patent application. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present application, several modifications and improvements can still be made, and these all belong to the protection scope of the present application. Therefore, the protection scope of the patent of the present application shall be subject to the appended claims.

Claims

1. An optical detection device, characterized in that: include: An optical fiber fixing seat, wherein the optical fiber fixing seat is provided with a plurality of optical fiber fixing holes arranged at intervals around a central axis, each of the optical fiber fixing holes being used for inserting an optical fiber; A rotating disk, rotatably connected to the optical fiber fixing seat, and the rotating disk can rotate around the central axis; A plurality of detection head assemblies are arranged on the rotating disk around the central axis, and the distance between any two adjacent detection head assemblies is greater than or equal to the distance between any two adjacent optical fiber fixing holes; the detection head assemblies can rotate with the rotating disk so that each of the detection head assemblies can be aligned with each of the optical fiber fixing holes in turn.

2. The optical detection device according to claim 1, characterized in that: The distance between two adjacent optical fiber fixing holes is greater than or equal to the aperture of the optical fiber fixing hole.

3. The optical detection device according to claim 1, characterized in that: The detection head assembly includes a shell, a light source, a first filter, a dichroic mirror, a second filter and a detector, wherein the light source, the first filter, the dichroic mirror, the second filter and the detector are arranged in the shell, and the light source, the first filter and the dichroic mirror are arranged at intervals along a first straight line; the dichroic mirror, the second filter and the detector are arranged at intervals along a second straight line perpendicular to the first straight line, and the dichroic mirror is arranged at an angle of 45° relative to the first straight line or the second straight line, so that the light emitted by the light source can be refracted in a direction away from the detector.

4. The optical detection device according to claim 3, characterized in that: A focusing lens is also arranged in the housing. The focusing lens is arranged on the second straight line at a side of the dichroic mirror away from the second filter.

5. The optical detection device according to claim 3, characterized in that: At least part of the shell is made of opaque material, and / or the shell is grounded.

6. The optical detection device according to claim 1, characterized in that: The optical detection device comprises a conductive slip ring and a cable. The conductive slip ring is coaxially mounted on the rotating disk. One end of the cable is connected to the detection head assembly, and the other end is connected to the conductive slip ring.

7. The optical detection device according to claim 1, characterized in that: A plurality of rolling balls are rollingly mounted on one side of the rotating disk facing the optical fiber fixing seat, and the rolling balls are rollingly connected to the optical fiber fixing seat.

8. The optical detection device according to claim 1, characterized in that: A driving element that is transmission-connected to the rotating disk is installed on the optical fiber fixing seat, and the driving element is used to drive the rotating disk to rotate; a sensing element is installed on one of the optical fiber fixing seat and the rotating disk, and a limiting plate is installed on the other, and the sensing element is configured to monitor the rotation state of the rotating disk driven by the driving element when it is aligned with the limiting plate.

9. The optical detection device according to claim 1, characterized in that: A plurality of mounting seats are installed on the edge of the rotating disk. The plurality of mounting seats are arranged at intervals around the central axis. Each of the detection head components is fixedly installed on a corresponding one of the mounting seats.

10. A PCR analyzer, characterized in that: It comprises a temperature-controlled amplification device and an optical detection device as described in any one of claims 1 to 9, wherein the optical detection device is connected to the temperature-controlled amplification device via an optical fiber.

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