A polymerase chain reaction (PCR) detector and its fluorescence detection device

Abstract

This application discloses a polymerase chain reaction (PCR) detector and its fluorescence detection device. The fluorescence detection device includes a sample reactor, an excitation component, and a fluorescence receiving tube, all fixedly mounted, as well as an excitation filter module, a detection filter module, and a control module, all driven by a driving device. The excitation filter sequentially passes through the optical path of the excitation component, and the corresponding detection filter simultaneously passes through the optical path of the fluorescence receiving tube. The fluorescence receiving tube receives fluorescence signals from at most one sample reactor at a time. This application can quickly, efficiently, and accurately complete multi-channel fluorescence detection, while also featuring a simple structure, small size, fast detection speed, high stability, and a significantly improved signal-to-noise ratio.

Description

Technical Field

[0001] This application belongs to the field of enzymology or microbiology device technology, and specifically relates to a polymerase chain reaction (PCR) detector and its included fluorescence detection device. Background Technology

[0002] Fluorescent polymerase chain reaction (PCR) is a high-precision molecular biology technique used to rapidly amplify and monitor the quantity of specific nucleic acid fragments in vitro, thereby enabling qualitative and quantitative analysis of target genes.

[0003] The fluorescence detection device in a polymerase chain reaction (PCR) analyzer should include a sample reactor, a light source tube for providing excitation light, and a fluorescence receiver tube for receiving corresponding fluorescence. The light source tube includes a light source and an excitation filter, and the receiver tube includes a detection filter and a photoelectric sensor. The light source tube for excitation light and the fluorescence receiver tube are configured accordingly. Different light source tubes can provide excitation light of different specific wavelengths to the sample reactor to excite the corresponding fluorescent probe in the reaction tube, causing it to emit fluorescence. The corresponding receiver tube is configured to receive the fluorescence, thereby realizing the detection of different fluorescence channels. The excitation filter in the excitation light source tube and the detection filter in the fluorescence receiver tube are also configured accordingly.

[0004] In existing technologies, when performing multi-channel fluorescence detection on the various sample reactors a1 (commonly 16, 48, or 96) of a polymerase chain reaction (PCR) instrument, to save costs and control equipment size, a dedicated light source tube and receiver tube are not configured for each sample reactor. Instead, multiple light source tubes a2 and receiver tubes a3, the same number as the fluorescence channels (commonly 4 or 8 channels), are fixed on a turntable a4. The turntable rotates, causing the light source tubes to sequentially align with the excitation fiber a6 to provide different wavelengths of excitation light to each sample reactor. The corresponding receiver tubes also rotate synchronously, sequentially aligning with the receiving fiber a5 to receive the fluorescence generated by each sample reactor. For example... Figure 1 As shown.

[0005] However, existing technical solutions still suffer from problems such as complex structure, large size, slow detection speed, low mechanical stability due to sudden stops and starts of the motor, poor alignment of the optical path and optical fiber, and short and limited integration time of the detection signal, resulting in a low signal-to-noise ratio. In order to solve these problems and improve the accuracy and efficiency of fluorescence detection, this invention is proposed. Summary of the Invention

[0006] To address the shortcomings or deficiencies of the prior art, this application provides a fluorescence detection device for a polymerase chain reaction (PCR) analyzer that is simple in structure, small in size, fast in detection speed, high in stability, and has a significantly improved signal-to-noise ratio, and provides a PCR analyzer including the fluorescence detection device.

[0007] This application is achieved through the following technical solution: A fluorescence detection device for a polymerase chain reaction (PCR) analyzer, comprising: The sample reactor, excitation assembly, and fluorescence receiver are all fixedly installed; a drive unit; and an excitation filter module and a detection filter module driven by the drive unit; and a control module.

[0008] Each of the sample reactors is connected to at least one excitation component and at least one fluorescence receiving tube via optical fiber; the sample reactor and the excitation component and the fluorescence receiving tube connected to the sample reactor via optical fiber constitute a detection unit.

[0009] The excitation assembly includes at least one excitation light source tube, and each excitation light source tube includes at least one light source; the fluorescence receiving tube includes a photoelectric sensor. When the excitation assembly includes multiple excitation light source tubes, each excitation light source tube may include a light source of different wavelengths, so that the excitation assembly can generate an excitation light source with a wider wavelength range.

[0010] The excitation filter module includes at least one excitation filter; the detection filter module includes at least one detection filter; the excitation filter and the detection filter used for the same fluorescence channel are referred to as corresponding filters.

[0011] The driving device drives the excitation filter module to move, so that the excitation filter passes through the optical paths of each excitation component in sequence; the driving device drives the detection filter module to move, so that the detection filter passes through the optical paths of each fluorescence receiving tube in sequence.

[0012] When the excitation filter passes through the excitation component optical path of a detection unit, the corresponding detection filter simultaneously passes through the fluorescence receiving tube optical path of the same detection unit; the fluorescence receiving tube receives at most one fluorescence signal emitted by the sample reactor at the same time.

[0013] The excitation component optical path refers to the optical path formed from the emission of a light beam from the light source until the light beam leaves the excitation source tube and enters the optical fiber. Since the excitation component includes one or more of the excitation source tubes, the excitation component optical path described in this application has the same meaning as the excitation source tube optical path in the excitation component. The fluorescence receiving tube optical path refers to the optical path formed from the emission of fluorescence leaving the optical fiber and entering the fluorescence receiving tube until the fluorescence enters the photoelectric sensor.

[0014] The sample reactor is used to hold the reaction tube, the excitation component is used to provide excitation light, and the fluorescence receiving tube is used to receive fluorescence signals. It should be emphasized that, unlike the prior art and general technical knowledge, the excitation light source tube and fluorescence receiving tube of the present invention do not include filters. The excitation filter module and the detection filter module are set separately to cooperate with the excitation component and the fluorescence receiving tube for fluorescence detection.

[0015] Preferably, in the detection unit, the sample reactor is optically connected to one of the excitation components and optically connected to one of the fluorescence receiving tubes; more preferably, the number of sample reactors, excitation components, and fluorescence receiving tubes connected by optical fibers is the same.

[0016] Preferably, in the detection unit, the sample reactor is connected to N optical fibers of the excitation components and N optical fibers of the fluorescence receiving tubes, where N ≥ 2.

[0017] Preferably, in the detection unit, the sample reactor is connected to one of the excitation components and N of the fluorescence receiving optical fibers, where N≥2.

[0018] Preferably, in the detection unit, the sample reactor is connected to N excitation components and one fluorescence receiving tube optical fiber, where N≥2.

[0019] Preferably, multiple detection units share the fluorescence receiving tube or the excitation component.

[0020] Preferably, the excitation assembly includes a plurality of excitation light source tubes, each of which includes a different light source.

[0021] Preferably, the light source is an LED.

[0022] Preferably, the excitation filter module and the detection filter module are integrated and driven synchronously by the same driving device; or, the excitation filter module and the detection filter module are separate and driven by different driving devices respectively.

[0023] Preferably, the excitation filter module includes multiple excitation filters, and the detection filter module includes multiple detection filters. The number of excitation filters and detection filters is equal to or a multiple of the number of fluorescence channels.

[0024] Preferably, the multiple excitation filters in the excitation filter module pass through multiple excitation component optical paths simultaneously, and the multiple detection filters in the detection filter module pass through multiple fluorescence receiving tube optical paths simultaneously.

[0025] Preferably, the excitation filter and the detection filter extend in the direction of motion.

[0026] Preferably, the excitation light source tube further includes an excitation light focusing lens.

[0027] Preferably, the excitation light focusing lens is a combination of multiple lenses.

[0028] Preferably, the fluorescence receiving tube further includes a fluorescence collimating lens and / or a fluorescence focusing lens.

[0029] Preferably, the photoelectric sensor is a photodiode.

[0030] Preferably, the excitation component and / or the fluorescence receiving tube are arranged in a circular pattern.

[0031] Preferably, the excitation components and / or the fluorescence receiving tubes are arranged in a straight line.

[0032] And a polymerase chain reaction (PCR) detector, including the fluorescence detection device as described above.

[0033] Compared with the prior art, this invention application has the following technical effects: Because only the filter module moves while other components are fixed, the overall structure is simple, small in size, fast, highly stable, and the signal-to-noise ratio is significantly improved. 1. The drive unit only needs to drive the filter module, which reduces the load and makes it move with low inertia and high speed, which has outstanding technical effect under the current detection requirements of rapid PCR; 2. Active devices do not require rotation and do not need to be configured with conductive slip rings, making them less susceptible to interference during detection and ensuring high stability; 3. Compared with the prior art, the optical path of the excitation component and the optical path of the fluorescence receiving tube of the present invention remain fixed, and there will be no misalignment or unstable alignment between the optical path and the optical fiber. The integration time of the detection signal is long, and the signal-to-noise ratio is greatly improved. The longer the filter moves in the direction of motion, the longer the integration time of the detection signal and the higher the signal-to-noise ratio.

[0034] The fluorescence detection device of the polymerase chain reaction detector described in this invention application can perform multi-channel fluorescence detection conveniently, efficiently, and accurately. Attached Figure Description

[0035] Other features, objects, and advantages of this application will become more apparent from the following detailed description of non-limiting embodiments with reference to the accompanying drawings: Figure 1 : Illustration of an existing fluorescence detection device; Figure 2 : A perspective view of an embodiment of the fluorescence detection device described in this application; Figure 3 : An exploded view of an embodiment of the fluorescence detection device described in this application; Figure 4 : A top view of an embodiment of the fluorescence detection device described in this application; Figure 5A : A top view of the rotating disk in Embodiment 1 of the fluorescence detection device described in this application Figure 1 ; Figure 5B : A top view of the rotating disk in Embodiment 1 of the fluorescence detection device described in this application Figure 2 (Continued) Figure 6 Cross-section of an embodiment of the fluorescence detection device described in this application. Figure 1 (Displays fluorescent receiver tube); Figure 7 Cross-section of an embodiment of the fluorescence detection device described in this application. Figure 2 (Displaying the excitation light source tube); Figure 8 Illustration of a filter module according to another embodiment of this application; Figure 9 : A perspective view of Embodiment 2 of the fluorescence detection device described in this application; Figure 10 : Exploded view of Embodiment 2 of the fluorescence detection device described in this application; Figure 11 Cross-section of Embodiment 2 of the fluorescence detection device described in this application. Figure 1 ; Figure 12 Cross-section of Embodiment 2 of the fluorescence detection device described in this application. Figure 2 ; Figure 13 : A perspective view of Embodiment 3 of the fluorescence detection device described in this application; Figure 14 : Exploded view of Embodiment 3 of the fluorescence detection device described in this application; Figure 15 Cross-section of Embodiment 3 of the fluorescence detection device described in this application. Figure 1 ; Figure 16 Cross-section of Embodiment 3 of the fluorescence detection device described in this application. Figure 2 ; Figure 17 : A schematic diagram of the detection unit of Embodiment 1 of the fluorescence detection device described in this application; Figure 18 : A schematic diagram of the detection unit of Embodiment 4 of the fluorescence detection device described in this application; Figure 19 : A schematic diagram of the detection unit of Embodiment 5 of the fluorescence detection device described in this application; Figure 20 : A schematic diagram of the detection unit of Embodiment Six of the fluorescence detection device described in this application; Figure 21: A schematic diagram of the detection unit of Embodiment 7 of the fluorescence detection device described in this application; 1 Sample reactor, 2 Excitation assembly, 3 Fluorescence receiving tube, 21 Excitation light source tube, 211 Light source, 212 Excitation light focusing lens, 31 Fluorescence collimating lens, 32 Fluorescence focusing lens, 33 Photoelectric sensor, 5 Drive device, 51 Excitation drive motor, 52 Detection drive motor, 6 Excitation filter module, 7 Detection filter module, 61 Excitation filter, 71 Detection filter, 8 Turntable, 81 Excitation turntable, 82 Detection turntable, 9 Lens disk, 10 Moving plate. Detailed Implementation

[0036] The technical solutions of this application will now be clearly and completely described with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.

[0037] Example 1 like Figures 2-8 , Figure 17 As shown, in this embodiment, the fluorescence detection device of the polymerase chain reaction (PCR) analyzer includes: Sixteen sample reactors 1, excitation components 2, and fluorescence receiver tubes 3 are fixedly installed. Each sample reactor 1 is connected to one excitation component 2 and one fluorescence receiver tube 3 via optical fiber, forming sixteen identical detection units. Sample reactor 1 is used to hold the reaction tubes. Figure 2 The image only shows the fiber optic connection of one of the sample reactors, 1. Each excitation assembly 2 includes four excitation light source tubes 21. Each excitation light source tube includes a light source 211 and a set of excitation light focusing lenses 212. The light sources 211 in the four excitation light source tubes 21 are: white LED, violet LED, deep red LED, and far-red LED. The white LED is used to excite the fluorescence of the ATTO425, FAM, HEX, ROX, and CY5 channels; the violet LED is used to excite the fluorescence of the AF405 channel; the deep red LED is used to excite the fluorescence of the QUASAR705 channel; and the far-red LED is used to excite the fluorescence of the CY7 channel.

[0038] Each fluorescence receiving tube 3 includes a fluorescence collimating lens 31, a fluorescence focusing lens 32, and a photoelectric sensor 33; in this embodiment, the photoelectric sensor 33 is preferably a photodiode.

[0039] The excitation focusing lens 212, as well as the fluorescence collimating lens 31 and fluorescence focusing lens 32, are fixed by a lens disk 9. The lens disk 9 has a central space to accommodate the passage of the turntable 8. Preferably, the lens disk 9 and all the lenses are integrally molded using two-color injection molding, and the lenses are made of transparent plastic. This significantly reduces the high economic and time costs associated with using traditional materials and processes due to the large number of lenses and the cumbersome gluing process.

[0040] like Figure 17 As shown, each sample reactor 1 is connected to four excitation light source tubes 21 in an excitation component 2 via four optical fibers, or a one-to-four fiber bundle can be used for connection; each sample reactor 1 is connected to a fluorescence receiving tube 3 via one optical fiber, forming a detection unit.

[0041] The excitation component 2 and the fluorescence receiving tube 3 are arranged in concentric circles.

[0042] The fluorescence detection device of the polymerase chain reaction (PCR) analyzer further includes a driving device 5, an excitation filter module 6, and a detection filter module 7. In this embodiment, the driving device 5 can be a drive motor, the excitation filter module 6 includes eight excitation filters 61, and the detection filter module 7 includes eight corresponding detection filters 71. Each excitation filter 61 and each detection filter 71 is considered a pair, and the eight pairs of filters correspond to the ATTO425, FAM, HEX, ROX, CY5, AF405, QUASAR705, and CY7 fluorescence channels, respectively. The eight excitation filters 61 and the eight detection filters 71 are arranged concentrically and fixedly on the same turntable 8, and the driving device 5 can drive the turntable 8 to rotate.

[0043] Unlike existing technologies, the sample reactor, excitation component, and fluorescence receiving tube in this invention are all in fixed positions. Furthermore, neither the excitation light source tube nor the fluorescence receiving tube includes filters. Through coordinated control by a control module, when the excitation filter corresponding to a specific fluorescence channel passes through the optical path of the excitation component in a detection unit, the detection filter corresponding to that specific fluorescence channel simultaneously passes through the optical path of the fluorescence receiving tube in the same detection unit, thus completing the detection of each fluorescence channel in each sample reactor. Since the active components do not rotate, the present invention eliminates the need for conductive slip rings, avoiding the risk of damage to conductive slip rings found in existing technologies. This results in a simple and stable overall structure for the fluorescence detection device.

[0044] The control module coordinates the opening and closing of the light source 211, the signal acquisition of the photoelectric sensor 33, and the movement of the drive device 5, which further drives the movement of the excitation filter module 6 and the detection filter module 7. The process of enabling the fluorescence detection device of the polymerase chain reaction detector to achieve detection through eight fluorescence channels is as follows: like Figure 4As shown, a sample reactor 1 (numbered A1, not shown) is selected, which is connected via optical fiber to four excitation light source tubes in an excitation assembly 2 (numbered G1), and via optical fiber to a fluorescence receiving tube 3 (numbered S1). Under the control of the control module, the drive device 5 drives the turntable 8 to rotate, causing the excitation filter 61 and the detection filter 71 to rotate; under the control of the control module, the excitation assembly 2 emits excitation light and forms an optical path.

[0045] First, when the ATTO425 excitation filter corresponding to the ATTO425 channel rotates under the drive of a motor through the optical path of the excitation source tube G1a in the excitation assembly G1, the ATTO425 detection filter simultaneously passes through the optical path of the fluorescence receiving tube S1. At the same time, the white LED in the excitation source tube G1a is activated, and the light beam passes through the excitation focusing lens 212 and the rotating ATTO425 excitation filter through this optical path, then leaves the excitation source tube G1a and reaches the sample reactor A1 via optical fiber. This excites a specific fluorescent marker in the reaction tube placed inside the sample reactor A1 to emit fluorescence. The fluorescence enters the fluorescence receiving tube S1 through optical fiber, passes through the fluorescence collimating lens 31, the ATTO425 detection filter, and the fluorescence focusing lens 32, and finally enters the photoelectric sensor 33. Thus, the sample reactor A1 completes the ATTO425 fluorescence channel detection.

[0046] Subsequently, as the FAM excitation filter corresponding to the FAM channel rotates with the drive motor and passes through the optical path of the excitation source tube G1a in the excitation assembly G1, the FAM detection filter simultaneously passes through the optical path of the fluorescence receiving tube S1. At the same time, the white LED of the excitation source tube G1a is activated, and the light beam passes through the excitation focusing lens 212, rotates through the FAM excitation filter in this optical path, and then reaches the sample reactor A1 through the optical fiber. This excites a specific fluorescent marker in the reaction tube placed inside the sample reactor A1 to emit fluorescence. The fluorescence enters the fluorescence receiving tube S1 through the optical fiber, passes through the fluorescence collimating lens 31, the FAM detection filter, and the fluorescence focusing lens 32, and finally enters the photoelectric sensor 33. Thus, the sample reactor A1 completes the FAM fluorescence channel detection.

[0047] Subsequently, as the HEX excitation filter corresponding to the HEX channel rotates with the drive motor through the optical path of the excitation source tube G1a in the excitation assembly G1, the HEX detection filter simultaneously passes through the optical path of the fluorescence receiving tube S1. At the same time, the white LED of the excitation source tube G1a is activated, and the light beam passes through the excitation focusing lens 212, rotates through the HEX excitation filter in this optical path, and then reaches the sample reactor A1 via optical fiber. This excites a specific fluorescent marker in the reaction tube placed inside the sample reactor A1 to emit fluorescence. The fluorescence enters the fluorescence receiving tube S1 through optical fiber, passes through the fluorescence collimating lens 31, the HEX detection filter, and the fluorescence focusing lens 32, and finally enters the photoelectric sensor 33. Thus, the sample reactor A1 completes the HEX fluorescence channel detection.

[0048] Subsequently, as the ROX excitation filter corresponding to the ROX channel rotates with the drive motor and passes through the optical path of the excitation source tube G1a in the excitation assembly G1, the ROX detection filter simultaneously passes through the optical path of the fluorescence receiving tube S1. At the same time, the white LED of the excitation source tube G1a is activated, and the light beam passes through the excitation focusing lens 212, rotates through the ROX excitation filter in this optical path, and then reaches the sample reactor A1 through the optical fiber. This excites a specific fluorescent marker in the reaction tube placed inside the sample reactor A1 to emit fluorescence. The fluorescence enters the fluorescence receiving tube S1 through the optical fiber, passes through the fluorescence collimating lens 31, the ROX detection filter, and the fluorescence focusing lens 32, and finally enters the photoelectric sensor 33. Thus, the sample reactor A1 completes the ROX fluorescence channel detection.

[0049] Subsequently, as the CY5 excitation filter corresponding to the CY5 channel rotates with the drive motor and passes through the optical path of the excitation source tube G1a in the excitation assembly G1, the CY5 detection filter simultaneously passes through the optical path of the fluorescence receiving tube S1. At the same time, the white LED of the excitation source tube G1a is activated, and the light beam passes through the excitation focusing lens 212, rotates through the CY5 excitation filter in this optical path, and then reaches the sample reactor A1 through the optical fiber. This excites a specific fluorescent marker in the reaction tube placed inside the sample reactor A1 to emit fluorescence. The fluorescence enters the fluorescence receiving tube S1 through the optical fiber, passes through the fluorescence collimating lens 31, the CY5 detection filter, and the fluorescence focusing lens 32, and finally enters the photoelectric sensor 33. Thus, the sample reactor A1 completes the CY5 fluorescence channel detection.

[0050] Subsequently, as the AF405 excitation filter corresponding to the AF405 channel rotates with the drive motor and passes through the optical path of the excitation source tube G1b in the excitation assembly G1, the AF405 detection filter simultaneously passes through the optical path of the fluorescence receiving tube S1. At the same time, the violet LED of the excitation source tube G1b is activated, and the light beam passes through the excitation focusing lens 212, rotates through the AF405 excitation filter, and then reaches the sample reactor A1 via optical fiber. This excites a specific fluorescent marker in the reaction tube placed inside the sample reactor A1 to emit fluorescence. The fluorescence enters the fluorescence receiving tube S1 through optical fiber, passes through the fluorescence collimating lens 31, the AF405 detection filter, and the fluorescence focusing lens 32, and finally enters the photoelectric sensor 33. Thus, the sample reactor A1 completes the AF405 fluorescence channel detection.

[0051] Subsequently, as the QUASAR705 excitation filter corresponding to the QUASAR705 channel rotates with the drive motor, it passes through the optical path of the excitation source tube G1c in the excitation assembly G1, while the QUASAR705 detection filter simultaneously passes through the optical path of the fluorescence receiving tube S1. At the same time, the deep red LED of the excitation source tube G1c is activated, and the light beam passes through the excitation focusing lens 212, rotates through the QUASAR705 excitation filter, and then reaches the sample reactor A1 via optical fiber. This excites a specific fluorescent marker in the reaction tube placed inside the sample reactor A1 to emit fluorescence. The fluorescence enters the fluorescence receiving tube S1 through optical fiber, passes through the fluorescence collimating lens 31, the QUASAR705 detection filter, and the fluorescence focusing lens 32, and finally enters the photoelectric sensor 33. Thus, the sample reactor A1 completes the QUASAR705 fluorescence channel detection.

[0052] Subsequently, as the CY7 excitation filter corresponding to the CY7 channel rotates with the drive motor and passes through the optical path of the excitation source tube G1d in the excitation assembly G1, the CY7 detection filter simultaneously passes through the optical path of the fluorescence receiving tube S1. At the same time, the far-red LED of the excitation source tube G1d is activated, and the light beam passes through the excitation focusing lens 212, rotates through the CY7 excitation filter in this optical path, and then reaches the sample reactor A1 through the optical fiber. This excites a specific fluorescent marker in the reaction tube placed inside the sample reactor A1 to emit fluorescence. The fluorescence enters the fluorescence receiving tube S1 through the optical fiber, passes through the fluorescence collimating lens 31, the CY7 detection filter, and the fluorescence focusing lens 32, and finally enters the photoelectric sensor 33. Thus, the sample reactor A1 completes the CY7 fluorescence channel detection.

[0053] As can be seen, when the turntable 8 rotates one revolution, all eight pairs of filters corresponding to the ATTO425, FAM, HEX, ROX, CY5, AF405, QUASAR705, and CY7 channels can pass through the optical path of the excitation light source tubes G1a to G1d and the fluorescence receiving tube S1 connected to the sample reactor A1 in sequence, and the sample reactor A1 completes the detection of the eight fluorescence channels in sequence.

[0054] Furthermore, in this embodiment, starting with the excitation component G1, excitation components 2 arranged sequentially in the opposite direction to the rotation direction of the turntable 8 are defined as G1-G16, and starting with the fluorescence receiving tube S1, fluorescence receiving tubes 3 arranged sequentially in the opposite direction to the rotation direction of the turntable 8 are defined as S1-S16, corresponding to sample reactors A1-A16 respectively. Figure 4 , Figure 5A As shown, the fluorescence detection device in this embodiment has eight excitation filters 61 and sixteen excitation components 2, eight detection filters 71 and sixteen fluorescence receiving tubes 3.

[0055] Multiple sample reactors 1 can simultaneously perform detection of different fluorescence channels based on different filters rotated to their respective detection units; for example, when the ATTO425 excitation filter passes through the optical path of the excitation component G1, the FAM, HEX, ROX, CY5, AF405, QUASAR705, and CY7 excitation filters are simultaneously passing through the optical paths of the excitation components G3, G5, G7, G9, G11, G13, and G15, respectively; and the ATTO425 detection filter is passing through the optical path of the fluorescence receiving tube S1, while the FAM, HEX, ROX, CY5, AF405, QUASAR705, and CY7 detection filters are simultaneously passing through the optical paths of the fluorescence receiving tubes S3, S5, S7, S9, S11, S13, and S15, respectively.

[0056] Therefore, when sample reactor A1 performs ATTO425 channel detection, sample reactor A3 simultaneously performs FAM channel detection, sample reactor A5 simultaneously performs HEX channel detection, sample reactor A7 simultaneously performs ROX detection, sample reactor A9 simultaneously performs CY5 channel detection, sample reactor A11 simultaneously performs AF405 channel detection, sample reactor A13 simultaneously performs QUASAR705 channel detection, and sample reactor A15 simultaneously performs CY7 channel detection.

[0057] Subsequently, when sample reactors A1, A3, A5, A7, A9, A11, A13, and A15 simultaneously completed one test, turntable 8 rotated to the position shown in the image. Figure 5BAs shown, the FAM excitation filter passes through the optical path of excitation component G2, and the FAM detection filter passes through the optical path of fluorescence receiving tube S2, allowing sample reactor A2 to perform FAM channel detection. At this time, the HEX, ROX, CY5, AF405, QUASAR705, CY7, and ATTO425 excitation filters simultaneously pass through the optical paths of excitation components G4, G6, G8, G10, G12, G14, and G16, respectively; the HEX, ROX, CY5, AF405, QUASAR705, CY7, and ATTO425 detection filters simultaneously pass through the optical paths of fluorescence receiving tubes S4, S6, S8, S10, S12, S14, and S16, respectively.

[0058] Therefore, when sample reactor A2 performs FAM channel detection, sample reactor A4 simultaneously performs HEX channel detection, sample reactor A6 simultaneously performs ROX detection, sample reactor A8 simultaneously performs CY5 channel detection, sample reactor A10 simultaneously performs AF405 channel detection, sample reactor A12 simultaneously performs QUASAR705 channel detection, sample reactor A14 simultaneously performs CY7 channel detection, and sample reactor A16 simultaneously performs ATTO425 channel detection. In this way, sample reactors A1 through A16 each complete one detection cycle.

[0059] As can be seen, when the turntable 8 rotates once, all sample reactors 1, namely sample reactors A1-A16, can complete the detection of eight fluorescence channels, and each fluorescence receiving tube 3 can receive at most one fluorescence signal emitted by the sample reactor 1 at the same time; thus, multi-channel fluorescence detection can be completed efficiently and accurately.

[0060] In existing technologies, the light source tube and fluorescence receiving tube rotate and stop at high speed with the motor. Each stop requires perfect alignment of the optical path with the fixedly installed optical fiber. This not only places extremely high demands on the precision of the motor but also makes it difficult to eliminate alignment errors. Even if alignment is achieved at a certain moment, the alignment time is extremely short and limited, resulting in very short periods of time for each light source tube to illuminate the sample reactor and for each fluorescence receiving tube to receive fluorescence. In this invention, because the excitation light source tube 21 is connected to the sample reactor 1 via optical fiber and its position is fixed, and the fluorescence receiving tube 3 is also connected to the sample reactor 1 via optical fiber and its position is fixed, the optical path of the excitation component and the optical path of the receiving tube are always aligned with the optical fiber. Only the filter passing through the optical path moves. The area of ​​the filter is larger than the cross-sectional area of ​​the optical path, completely eliminating the above-mentioned problems. At the same time, during the time the filter passes through the optical path, the excitation light source tube 21 can illuminate the sample reactor 1, and the fluorescence receiving tube 3 can synchronously receive the fluorescence signal. The longer the filter passes through the optical path, the longer the sample reactor 1 is illuminated by the excitation light, the longer the integration time of the detection signal, and the signal-to-noise ratio will be significantly improved.

[0061] Therefore, preferably, in this embodiment, the lengths of the excitation filter 61 and the detection filter 71 are extendable in their rotational direction. Specifically, in this embodiment, both the excitation filter 61 and the detection filter 71 can be lengthened along their direction of movement, extending as far as their adjacent filters. Each excitation filter 61 can extend to occupy 1 / 8 of its circumference, and each detection filter 71 can extend to occupy 1 / 8 of its circumference, such as... Figure 8 As shown. In other embodiments, the maximum proportion of the extended excitation filter 61 and detection filter 71 occupying a single loop can vary depending on the number of filters.

[0062] Preferably, the difference between Embodiment 1A and Embodiment 1 is that the fluorescence detection device still has eight fluorescence channels, but the excitation filter module 6 is configured to include sixteen (or other multiples of eight) excitation filters 61, and the detection filter module 7 is configured to include sixteen (or other multiples of eight) corresponding detection filters 71. All filters are divided into two groups, with eight pairs in each group, corresponding to eight fluorescence channels. In this embodiment, under the coordinated control of the control module, the excitation filter 61 / detection filter 71 only needs to rotate half a turn for all sample reactors 1 to complete one detection of eight fluorescence channels. When rotated one turn, the sample reactors 1 can complete two detections of eight fluorescence channels, greatly improving the detection efficiency. Similarly, depending on different needs, the number of filters can be set to other multiples of the number of fluorescence channels.

[0063] The difference between Example 1B and Example 1 is that the number of excitation filters 61 and detection filters 71 is set to sixteen, meaning each fluorescence channel corresponds to two identical excitation filters 61 and two identical detection filters 71. Based on this arrangement, under the coordinated control of the control module, sample reactors A1 and A2 can simultaneously perform ATTO425 channel detection. At this time, sample reactors A3 and A4 simultaneously perform FAM channel detection, sample reactors A5 and A6 simultaneously perform HEX channel detection, sample reactors A7 and A8 simultaneously perform ROX detection, sample reactors A9 and A10 simultaneously perform CY5 channel detection, sample reactors A11 and A12 simultaneously perform AF405 channel detection, sample reactors A13 and A14 simultaneously perform QUASAR705 channel detection, and sample reactors A15 and A16 simultaneously perform CY7 channel detection. Sample reactors A1-A16 can complete one detection simultaneously.

[0064] The difference between Example 1C and Example 1 is that the two excitation light source tubes 21 arranged along the rotation direction in the excitation assembly 2 (e.g., Figure 4Each of the light source tubes c and d in the excitation assembly 2 includes a white LED; thus, the length of the excitation filter 61 does not need to be changed. For example, as the ATTO425 excitation filter rotates through the optical path of the two excitation light source tubes, the control module simultaneously coordinates and controls the two white LEDs to emit light sequentially, doubling the integration time of the ATTO425 channel signal, thereby significantly improving the detection efficiency of the fluorescence channel. The ATTO425 excitation filter can also be a FAM excitation filter, a HEX excitation filter, a ROX excitation filter, or a CY5 excitation filter. In other embodiments, the light sources 211 in the two excitation light source tubes 21 arranged along the rotation direction in the excitation assembly 2 can also be two identical: a violet LED with a corresponding AF405 excitation filter; or a deep red LED with a corresponding QUASAR705 excitation filter; or a far-red LED with a corresponding CY7 excitation filter.

[0065] The difference between Example 1D and Example 1 is that the excitation component 2 is configured to include only one excitation light source tube 21, which includes a full-spectrum LED, a multi-color LED or a white LED. The filters are in multiple sets. Under the coordinated control of the control module, the excitation light source tube 21 forms the excitation light required for each fluorescence channel by combining with different excitation filters 61. The excitation component 2 can still provide excitation light of different wavelengths.

[0066] In other embodiments, the excitation component 2 may include only one excitation light source tube 21, which may include only one ordinary light source, such as a white LED. In this case, if the filter is only one set, the excitation component 2 and the excitation filter 61 can only provide one type of excitation light. Although each sample reactor 1 only performs single-channel fluorescence detection, it still retains the technical effect described in this invention and should be covered within the scope of this invention.

[0067] Example 2 like Figures 9-12 As shown, the difference from Embodiment 1 is that in this embodiment, two sets of drive motors and their driven turntables are provided. The excitation filter module 6 is fixedly arranged on the excitation turntable 81, and the detection filter module 7 is fixedly arranged on the detection turntable 82. The drive device 5 includes an excitation drive motor 51 and a detection drive motor 52. The excitation drive motor 51 drives the excitation turntable 81 to rotate, causing the excitation filter 61 to rotate; the detection drive motor 52 drives the detection turntable 82 to rotate, causing the detection filter 71 to rotate. Under the coordinated control of the control module, when the excitation filter 61 of a specific fluorescence channel passes through the optical path of the excitation component of a detection unit, the detection filter 71 corresponding to that fluorescence channel also synchronously passes through the optical path of the fluorescence receiving tube of the same detection unit.

[0068] Example 3 like Figures 13-16 As shown, the difference from Embodiment 1 is that in this embodiment, the excitation component 2 and the fluorescence receiving tube 3 are arranged in a straight line, and the excitation filter module 6 and the detection filter module 7 are both mounted on the shift plate 10, which can move linearly under the drive of the driving device 5. Under the coordinated control of the control module, when the excitation filter 61 of a specific fluorescence channel passes through the optical path of the excitation component of a certain detection unit, the detection filter 71 corresponding to that fluorescence channel also simultaneously passes through the optical path of the fluorescence receiving tube of the same detection unit.

[0069] In this embodiment, the maximum unidirectional linear stroke of the shift plate 10 should ensure that all excitation filters 61 can pass through the optical paths of all excitation light source tubes of all excitation components 2; and that all detection filters 71 can pass through the optical paths of all fluorescence receiving tubes. When the shift plate 10 completes one maximum unidirectional linear stroke, all sample reactors 1 can complete the detection of all fluorescence channels.

[0070] In this embodiment, the lengths of the excitation filter 61 and the detection filter 71 extend in their linear motion direction. Preferably, the length of the excitation filter 61 can extend to its adjacent excitation filter, and the length of the detection filter 71 can extend to its adjacent detection filter.

[0071] Example 4 like Figure 18 As shown, the difference from Embodiment 1 is that in this embodiment, the number of sample reactors 1 is sixteen, and the number of excitation components 2 and fluorescence receiving tubes 3 is set to thirty-two; in each detection unit, the sample reactor 1 is optically connected to two excitation components 2 and two fluorescence receiving tubes 3 respectively.

[0072] In one operation of the detection unit, two excitation components 2 are arranged adjacently on their circumference. Under the coordinated control of the control module, they cooperate with the excitation filter 61 to emit the same fluorescence channel excitation light to the same sample reactor 1. Two fluorescence receiving tubes 3 successively receive and detect the same fluorescence channel signal emitted by the same sample reactor 1. Based on the teachings of this application, those skilled in the art will understand that the excitation filter 61 and the detection filter 71 are adjusted and set accordingly on the turntable 8. Those skilled in the art will understand that because the number of excitation components and fluorescence receiving tubes is doubled, the excitation filter module 6 and the detection filter module 7 can perform two detections of all fluorescence channels on all sample reactors for each rotation, doubling the integration time and the signal-to-noise ratio.

[0073] In another operation of the detection unit, two excitation components 2 are arranged 180 degrees apart on their circumferences, and two fluorescence receiving tubes 3 are also arranged 180 degrees apart on their circumferences. Based on the teachings of this application, those skilled in the art will understand that the excitation filter 61 and the detection filter 71 are adjusted and set accordingly on the turntable 8; for example, the excitation filter 61 and the detection filter 71 are arranged continuously on a half-circle turntable 8. Under the coordinated control of the control module, the two excitation components 2 and the excitation filter 61 cooperate to emit different fluorescence channel excitation light to the sample reactor 1 sequentially, and the two fluorescence receiving tubes 3 sequentially receive and detect the different fluorescence channel signals emitted by the sample reactor 1. With this setup, the excitation filter module 6 and the detection filter module 7 only need to rotate half a circle to perform one detection of all fluorescence channels in all sample reactors 1, doubling the detection resolution; continuing to rotate half a circle allows for two detections of all fluorescence channels in all sample reactors 1, doubling the integration time. Furthermore, when the fluorescence detection device periodically activates sample reactor 1 via the control program, sample reactor 1 only needs to wait for the turntable 8 to rotate half a revolution before it can begin detection, eliminating the need to wait a full week. This improves the detection speed and efficiency for achieving rapid PCR.

[0074] Similarly, the number of excitation components and fluorescence receiving tubes can be N times the number of sample reactors, where N can be an integer of 2 or greater.

[0075] Example 5 like Figure 19 As shown, the difference from Embodiment 1 is that in this embodiment, the number of sample reactors 1 is sixteen, the number of excitation components 2 is set to eight, and the number of fluorescence receiving tubes 3 is set to sixteen; all sample reactors 1 are optically connected to one excitation component 2 and one fluorescence receiving tube 3 respectively; each excitation component 2 is connected to two sample reactors 1 through an optical fiber, that is, two detection units share one excitation component 2.

[0076] In the operation of the detection unit with two shared excitation components 2 described in this embodiment, under the coordinated control of the control module, the same excitation component 2 and the excitation filter 61 simultaneously emit the same fluorescence channel excitation light to the two sample reactors 1. The fluorescence receiving tubes 3 connected to each of the two sample reactors 1 simultaneously receive the same fluorescence channel signal emitted by the two sample reactors 1. Based on the teachings of this application, those skilled in the art will understand that the excitation filter 61 and the detection filter 71 are adjusted and set accordingly on the turntable 8.

[0077] In this embodiment, because the number of excitation components 2 is halved, the use of light source 211 is greatly reduced, which can significantly reduce costs. Similarly, in other embodiments, the number of fluorescence receiving tubes 3 and sample reactors 1 can be N times the number of excitation components.

[0078] Example 6 like Figure 20 As shown, the difference from Embodiment 1 is that in this embodiment, the number of sample reactors 1 is sixteen, the number of excitation components 2 is set to sixteen, and the number of fluorescence receiving tubes 3 is set to eight; all sample reactors 1 are optically connected to one excitation component 2 and one fluorescence receiving tube 3 respectively; each fluorescence receiving tube 3 is connected to two sample reactors 1 through optical fiber, that is, two detection units share one fluorescence receiving tube 3.

[0079] In the operation of the detection unit described in this embodiment, the control module coordinates and controls the two excitation components 2 and the excitation filter 61 to successively emit fluorescence channel excitation light to their respective connected sample reactors 1. The fluorescence receiving tube 3 shared by the two sample reactors 1 successively receives the fluorescence channel signals emitted by the two sample reactors 1. Based on the teachings of this application, those skilled in the art will understand that the excitation filter 61 and the detection filter 71 are adjusted and set accordingly on the turntable 8. In this embodiment, because the fluorescence receiving tube 3 is halved, the use of the photoelectric sensor 33 is greatly reduced, which can significantly reduce costs.

[0080] In the detection unit sharing a fluorescence receiving tube 3, each fluorescence receiving tube 3 receives at most one fluorescence signal emitted by the sample reactor 1 at any given time. Therefore, regardless of whether the fluorescence receiving tube 3 is shared, this application ensures that each fluorescence receiving tube 3 receives at most one fluorescence signal emitted by the sample reactor 1 at any given time. Compared to the prior art scheme of sharing a fluorescence receiving tube and simultaneously receiving array signals through a CCD camera, this overcomes the problems of pixel aliasing and signal crosstalk, significantly improving the accuracy and signal-to-noise ratio of the detection results.

[0081] Example 7 like Figure 21 As shown, the difference from Embodiment 1 is that in this embodiment, the number of sample reactors 1 is sixteen, the number of excitation components 2 is set to sixteen, and the number of fluorescence receiving tubes 3 is set to thirty-two; all sample reactors 1 are respectively connected to one excitation component 2 and two fluorescence receiving tubes 3 by optical fiber.

[0082] During the operation of the detection unit described in this embodiment, under the coordinated control of the control module, the two fluorescence receiving tubes 3 can simultaneously receive fluorescence signals from the same sample reactor 1, thereby increasing the integration time. Based on the teachings of this application, those skilled in the art will understand that the excitation filter 61 and the detection filter 71 are adjusted and set accordingly on the turntable 8.

[0083] Similarly, the number of excitation components 2 and fluorescence receiving tubes 3 can be N times the number of sample reactors 1, where N can be an integer of 2 or greater.

[0084] The above embodiments are only used to illustrate the technical solutions of this application and are not intended to limit it. The preferred embodiments have been described in detail. Those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of this application without departing from the spirit and scope of the technical solutions of this application, and all such modifications and substitutions should be covered within the scope of the claims of this application.

Claims

1. A fluorescence detection device for a polymerase chain reaction detector, characterized by, include: The sample reactor, excitation assembly, and fluorescence receiver are all fixedly installed; the drive unit, and the excitation filter module and detection filter module driven by the drive unit; the control module; Each of the sample reactors is connected to at least one excitation component and at least one fluorescence receiving tube via optical fiber; the sample reactor and the excitation component and fluorescence receiving tube connected to the sample reactor via optical fiber constitute a detection unit; The excitation assembly includes at least one excitation light source tube, and each excitation light source tube includes at least one light source; the fluorescence receiving tube includes a photoelectric sensor; in the detection unit, the sample reactor is connected to each of the excitation light source tubes in the excitation assembly via an optical fiber; The excitation filter module includes at least one excitation filter; the detection filter module includes at least one detection filter; the driving device drives the excitation filter module to move, so that the excitation filter passes through the optical paths of each excitation component in sequence; the driving device drives the detection filter module to move, so that the detection filter passes through the optical paths of each fluorescence receiving tube in sequence. When the excitation filter passes through the excitation component optical path of a detection unit, the corresponding detection filter simultaneously passes through the fluorescence receiving tube optical path of the same detection unit; the fluorescence receiving tube receives at most one fluorescence signal emitted by the sample reactor at the same time.

2. The fluorescence detection device of the polymerase chain reaction (PCR) detector according to claim 1, characterized in that: In the detection unit, the sample reactor is connected to an excitation component optical fiber and to a fluorescence receiver optical fiber.

3. The fluorescence detection device of the polymerase chain reaction detector according to claim 1, characterized in that: In the detection unit, the sample reactor is connected to N optical fibers of the excitation components and to N optical fibers of the fluorescence receiving tubes, where N≥2.

4. The fluorescence detection device of the polymerase chain reaction detector according to claim 1, characterized in that: In the detection unit, the sample reactor is connected to one of the excitation components by optical fiber and to N of the fluorescence receiving tubes by optical fiber, where N≥2.

5. The fluorescence detection device of the polymerase chain reaction (PCR) detector according to claim 1, characterized in that: In the detection unit, the sample reactor is optically connected to N excitation components and optically connected to one fluorescence receiving tube, where N≥2.

6. The fluorescence detection device of the polymerase chain reaction (PCR) detector according to any one of claims 2-5, characterized in that: Multiple detection units may share the fluorescence receiving tube or the excitation assembly.

7. The fluorescence detection device of the polymerase chain reaction (PCR) detector according to claim 2, characterized in that: The number of sample reactors, excitation components, and fluorescence receivers connected by optical fibers is the same.

8. The fluorescence detection device of the polymerase chain reaction detector according to claim 6, characterized in that: The excitation assembly includes multiple excitation light source tubes, each of which includes a different light source.

9. The fluorescence detection device of the polymerase chain reaction detector according to claim 1, characterized in that: The light source is an LED.

10. The fluorescence detection device of the polymerase chain reaction detector according to claim 1, characterized in that: The excitation filter module and the detection filter module are integrated and driven synchronously by the same driving device.

11. The fluorescence detection device of the polymerase chain reaction detector according to claim 1, characterized in that: The excitation filter module and the detection filter module are separate and are driven by different driving devices.

12. The fluorescence detection device of the polymerase chain reaction detector according to claim 1, characterized in that: The excitation filter module includes multiple excitation filters, and the detection filter module includes multiple detection filters. The number of excitation filters and detection filters is equal to or a multiple of the number of fluorescence channels.

13. The fluorescence detection device of the polymerase chain reaction detector according to claim 12, characterized in that: The multiple excitation filters in the excitation filter module pass through multiple excitation component optical paths simultaneously, and the multiple detection filters in the detection filter module pass through multiple fluorescence receiving tube optical paths simultaneously.

14. The fluorescence detection device of the polymerase chain reaction detector according to claim 1, characterized in that: The lengths of the excitation filter and the detection filter extend in their direction of motion.

15. The fluorescence detection device of the polymerase chain reaction detector according to claim 6, characterized in that: The excitation light source tube also includes an excitation light focusing lens.

16. The fluorescence detection device of the polymerase chain reaction detector according to claim 15, characterized in that: The excitation light focusing lens is a combination of multiple lenses.

17. The fluorescence detection device of the polymerase chain reaction detector according to claim 1, characterized in that: The photoelectric sensor is a photodiode.

18. The fluorescence detection device of the polymerase chain reaction detector according to claim 1, characterized in that: The fluorescence receiving tube also includes a fluorescence collimating lens and / or a fluorescence focusing lens.

19. The fluorescence detection device of the polymerase chain reaction detector according to claim 1, characterized in that: The excitation components and / or the fluorescence receiving tubes are arranged in a circular pattern.

20. The fluorescence detection device of the polymerase chain reaction detector according to claim 1, characterized in that: The excitation components and / or the fluorescence receiving tubes are arranged in a straight line.

21. A polymerase chain reaction (PCR) detector, comprising a fluorescence detection device, characterized in that: The fluorescence detection device is the fluorescence detection device according to any one of claims 1-20.

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