Two-color fluorescence detection assembly and POCT (Point of Care Testing) molecular diagnosis equipment

Abstract

The utility model discloses a two-color fluorescence detection assembly and POCT (Point of Care Testing) molecular diagnosis equipment. A bi-color fluorescence detection assembly includes a single excitation light source having a wavelength range configured to excite two dyes within a reaction tube to generate fluorescence emission light having different wavelength ranges, a color sensor configured to sense the fluorescence emission light having different wavelength ranges, and a color sensor configured to sense the fluorescence emission light having different wavelength ranges. The fluorescence detector is configured to be capable of transmitting fluorescence of two dyes in the fluorescence emission light and converting the two fluorescence into electric signals respectively; and the control module is connected with the color sensor, and the control module is configured to process the electric signal to obtain a two-color fluorescence output value. According to the POCT molecular diagnosis equipment, complex light path design is not needed, double-color fluorescence detection can be completed only by arranging the single excitation light source and the color sensor, and the POCT molecular diagnosis equipment is simple in structure, low in cost, capable of being integrated in micro equipment and easy to carry.

Description

Technical Field

[0001] The present disclosure relates to the technical field of intelligent detection devices, and particularly to a dual-color fluorescence detection component and a POCT molecular diagnostic device. Background Art

[0002] The commonly used detection methods of existing POCT molecular diagnostic devices mainly include colorimetry, electrochemistry, and fluorescence. These methods have their own advantages and disadvantages. The advantage of colorimetry is simple operation and low cost, but the detection sensitivity is low and the results are greatly interfered. The advantage of electrochemistry is high sensitivity, but the electrode cost is high, it is easily contaminated, and the result accuracy is poor. Compared with the previous two methods, fluorescence has higher sensitivity and specificity, and can realize real-time quantitative detection, and can be made into fluorescence probes to further increase the detection specificity. However, the limitation of monochromatic fluorescence is that it can only detect a single molecule and cannot detect multiple molecules simultaneously.

[0003] Therefore, multi-color fluorescence is widely used in the field of molecular diagnosis in order to detect multiple molecules simultaneously and improve the detection sensitivity and specificity. Currently, the traditional multi-channel fluorescence detection uses the method of physically switching the excitation light and the filter. However, this method has a disadvantage that a moving component needs to be added to achieve the moving switching or rotational switching of the excitation light and the filter, which makes it difficult to integrate in small devices. Among them, dual-color fluorescence detection is mostly used in multi-color fluorescence detection, and the dual-color fluorescence detection method can use one sample for dual-index detection, saving samples, increasing the detection throughput, saving device space, and facilitating the setting of detection internal references. The traditional implementation method of the dual-fluorescence channel is a complex optical path design, which is achieved by multiple light sources plus multiple groups of filters. Its disadvantages are high cost, the complex optical path leads to high requirements for processing accuracy, large processing difficulty, high maintenance cost, large volume, and it cannot be integrated into small devices. Therefore, although the dual-color fluorescence detection method has great application potential, due to the complexity of its optical path, there is currently no extremely miniaturized (<35 g / detection hole), low-cost POCT molecular diagnostic device for dual-color fluorescence detection. Summary of the Utility Model

[0004] The present disclosure provides a dual-color fluorescence detection component and a POCT molecular diagnostic device to at least solve one of the technical problems existing in the prior art.

[0005] According to a first aspect of the present disclosure, there is provided a dual-color fluorescence detection component, including,

[0006] A single excitation light source having a wavelength range, which is configured to be able to excite two dyes in a reaction tube to generate fluorescence emission light having different wavelength ranges.

[0007] A color sensor configured to be able to transmit the fluorescence of two dyes in the fluorescence emission light and convert the two fluorescences into electrical signals respectively;

[0008] A control module connected to the color sensor, the control module being configured to process the electrical signals to obtain a dual-color fluorescence output value.

[0009] In an implementable embodiment, the color sensor has a filter array, the filter array at least includes a first filter and a second filter and the two have different wavelength channels, the first filter is configured to be able to transmit the fluorescence of the first dye in the fluorescence emission light and not transmit the fluorescence of the second dye, and the second filter is configured to be able to transmit the fluorescence of the second dye in the fluorescence emission light and not transmit the fluorescence of the first dye.

[0010] In an implementable embodiment, the transmission wavelength of the first filter is the maximum fluorescence emission wavelength of the first dye ±10 - 25 nm;

[0011] The transmission wavelength of the second filter is the maximum fluorescence emission wavelength of the second dye ±10 - 25 nm.

[0012] In an implementable embodiment, the color sensor is an RGB color sensor, the RGB color sensor has a filter array, the filter array at least includes a first filter and a second filter, the first filter is used to transmit the G fluorescence of the first dye in the fluorescence emission light, and the second filter is used to transmit the R fluorescence of the second dye in the fluorescence emission light; the RGB sensor converts the transmitted G fluorescence and R fluorescence into electrical signals respectively through the filter array it has; the control module collects the electrical signals in real time and processes them to obtain Δ G, ΔR dual-color fluorescence output values.

[0013] In an implementable embodiment, the color sensor is an RGB color sensor, the RGB color sensor has a filter array, the filter array at least includes a first filter and a second filter, the first filter is used to transmit the G fluorescence of the two dyes in the fluorescence emission light, and the second filter is used to transmit the R fluorescence of the two dyes in the fluorescence emission light; the RGB sensor converts the transmitted G fluorescence and R fluorescence into electrical signals respectively through the filter array it has; the control module collects the electrical signals in real time and processes them to obtain Δ G, ΔR dual-color fluorescence output values.

[0014] In an implementable embodiment, when the color sensor is an RGB color sensor, the fluorescent dyes used are configured to be able to generate fluorescence with a light wavelength of 390 - 780 nm.

[0015] In one implementable embodiment, the single excitation light source uses blue light or ultraviolet light, and the wavelength range of the single excitation light source is 260-500 nm.

[0016] In one implementable embodiment, the wavelength range of the single excitation light source is 380-480 nm.

[0017] In one implementable embodiment, a high-pass filter is further included. The high-pass filter is disposed in front of the color sensor and is used to first filter the excitation light in the fluorescence emission light and transmit the filtered fluorescence emission light to the color sensor.

[0018] The color sensor, the high-pass filter, and the reaction tube are arranged in one-to-one correspondence.

[0019] In one implementable embodiment, the color sensor is configured to be able to filter the excitation light in the fluorescence emission light while transmitting the fluorescence of two dyes in the fluorescence emission light and convert the two fluorescences into electrical signals respectively.

[0020] According to a second aspect of the present disclosure, a POCT molecular diagnostic device is provided, which includes the dual-color fluorescence detection component in any one of the implementable embodiments of the first aspect.

[0021] Compared with the prior art, the advantages of the present application are as follows: The POCT molecular diagnostic device of the present application does not require a complex optical path design. Only by setting a single excitation light source and a color sensor can dual-color fluorescence detection be completed. It has a simple structure, low cost, can be integrated into a micro device, and is easy to carry. In the present application, by using an RGB array sensor to collect data of multiple points in a small range and cooperating with the processing method of the control module, the fluorescence values of different wavelengths corresponding to two target analytes can be clearly confirmed, so that the dual-color fluorescence output value and the concentration contents of the two target analytes can be accurately obtained.

[0022] It should be understood that the content described in this part is not intended to identify the key or important features of the embodiments of the present disclosure, nor is it used to limit the scope of the present disclosure. Other features of the present disclosure will become easily understood through the following description. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] By reading the following detailed description with reference to the accompanying drawings, the above and other objects, features, and advantages of the exemplary embodiments of the present disclosure will become easily understood. In the drawings, several embodiments of the present disclosure are shown in an exemplary rather than restrictive manner, wherein:

[0024] In the drawings, the same or corresponding reference numerals represent the same or corresponding parts.

[0025] Figure 1Shows the structural schematic of the POCT molecular diagnostic device according to an embodiment of the present disclosure Figure 1 ;

[0026] Figure 2 Shows the structural schematic of the POCT molecular diagnostic device according to an embodiment of the present disclosure Figure 2 ;

[0027] Figure 3 Shows the structural schematic of the POCT molecular diagnostic device according to an embodiment of the present disclosure Figure 3 ;

[0028] Figure 4 Shows the structural schematic of the POCT molecular diagnostic device according to an embodiment of the present disclosure Figure 4 ;

[0029] Figure 5 Shows the structural schematic diagram of the RGB sensor according to an embodiment of the present disclosure;

[0030] Figure 6 Shows the installation position schematic of the single excitation light source and the reaction tube according to an embodiment of the present disclosure Figure 1 ;

[0031] Figure 7 Shows the installation position schematic of the single excitation light source and the reaction tube according to an embodiment of the present disclosure Figure 2 ;

[0032] Figure 8 Shows the installation position schematic of the single excitation light source and the reaction tube according to an embodiment of the present disclosure Figure 3 ;

[0033] Figure 9 Shows the installation position schematic of the single excitation light source and the reaction tube according to an embodiment of the present disclosure Figure 4 ;

[0034] Figure 10 Shows the structural schematic of another POCT molecular diagnostic device according to an embodiment of the present disclosure Figure 1 ;

[0035] Figure 11 Shows the structural schematic of another POCT molecular diagnostic device according to an embodiment of the present disclosure Figure 2 ;

[0036] Figure 12 Shows the structural schematic of another POCT molecular diagnostic device according to an embodiment of the present disclosure Figure 3 .

[0037] Description of the reference numerals in the figure: Carrier module 1, RGB array sensor 2, RGB array sensor mounting substrate 3, filter mounting substrate 4, high-pass filter 5, single excitation light source 6, control module 7, first observation hole 11. Detailed implementation manners

[0038] To make the objectives, features, and advantages of the present disclosure more obvious and understandable, the technical solutions in the embodiments of the present disclosure will be clearly and completely described below in conjunction with the accompanying drawings in the embodiments of the present disclosure. Obviously, the described embodiments are only a part of the embodiments of the present disclosure, rather than all the embodiments. Based on the embodiments in the present disclosure, all other embodiments obtained by those skilled in the art without creative efforts belong to the scope of protection of the present disclosure.

[0039] Since the current traditional dual-color fluorescence detection requires complex optical path design, that is, multiple light sources are configured with multiple sets of filter films, resulting in a series of disadvantages such as high processing difficulty, high maintenance cost, large volume of the equipment with such a complex optical path, inability to be integrated into small-volume detection equipment, and inconvenience in carrying. We have developed a new dual-color fluorescence detection method and can be integrated into a micro molecular diagnostic device.

[0040] An RGB array sensor (also known as an RGB color sensor, RGB sensor) is a sensor that integrates a filter film array and an optoelectronic device array with red (R), green (G), and blue (B) filter channels, and can convert the captured optical signal into an electrical signal. These optoelectronic devices are usually photoresistors, photodiodes (photoelectric diodes), phototransistors, etc. They are arranged in an array, and the detection signal wavelength of each point is selected by covering a filter film with different wavelength channels (a color filter matrix) on the corresponding point of each array. Therefore, optical signals of different colors can be sensed simultaneously. The RGB array sensor has the advantages of small volume and low price, and can realize data acquisition of multiple points in a small range.

[0041] Therefore, we found that when the RGB array sensor is applied to a molecular POCT detection device, dual-color fluorescence output values can be obtained.

[0042] Based on this, according to an embodiment of the present disclosure, the present application provides a dual-color fluorescence detection component.

[0043] A dual-color fluorescence detection component includes a single excitation light source having a wavelength range, which is configured to be able to excite two dyes in a reaction tube to generate fluorescence emission light having different wavelength ranges.

[0044] A color sensor, which is configured to be able to transmit the fluorescence of the two dyes in the fluorescence emission light and convert the two fluorescences into electrical signals respectively.

[0045] A control module, connected to the color sensor, and the control module is configured to be able to process the received electrical signals to obtain dual-color fluorescence output values.

[0046] For example, in the POCT molecular diagnostic device of the present application, generally multiple reaction tubes are provided (it is also possible to provide one, and correspondingly one color sensor is provided. The number of reaction tubes in the present application is not specifically limited). Correspondingly, multiple color sensors are provided. The number of color sensors is set corresponding to the number of reaction tubes, and their installation positions also correspond to each other. By providing a single excitation light source in the present application, the single excitation light source can simultaneously excite two dyes (fluorescent dyes) in the reaction tube, causing the two fluorescent dyes to generate fluorescent emission lights with different wavelength ranges. And by providing a color sensor, the color sensor can transmit the fluorescence of the two dyes in the fluorescent emission light and convert the two fluorescences into electrical signals respectively (that is, the color sensor is configured to be able to transmit the fluorescence within the wavelength range of the fluorescent emission lights of the two dyes in the fluorescent emission light and convert the fluorescence of these two wavelengths into electrical signals respectively), and then transmit the electrical signals to the control module, and the control module processes the electrical signals to obtain a dual-color fluorescence output value.

[0047] Therefore, the POCT molecular diagnostic device of the present application does not require a complex optical path design. Only by providing a single excitation light source and a color sensor can dual-color fluorescence detection be completed. It has a simple structure, can be integrated into a micro device, and is easy to carry.

[0048] In one embodiment, the color sensor generally mainly includes a filter array and a photoelectric device. The filter array is used to transmit light of a specific wavelength in the emission light, and then the photoelectric device performs photoelectric conversion. Therefore, the color sensor of the present application has a filter array. The filter array includes at least a first filter and a second filter, and the two have different wavelength channels. The first filter is configured to be able to transmit the fluorescence of the first dye in the fluorescent emission light and not be able to transmit the fluorescence of the second dye. The second filter is configured to be able to transmit the fluorescence of the second dye in the fluorescent emission light and not be able to transmit the fluorescence of the first dye. The fluorescence of the two dyes in the fluorescent emission light output from the reaction tube is transmitted through the filter array to achieve the purpose of separating the fluorescence of the two dyes. Then, the two fluorescences are respectively converted into electrical signals by the photoelectric device carried by the color sensor, and the control module processes the received electrical signals to obtain a dual-color fluorescence output value. In addition, the control module can also be connected to a terminal, and the terminal can draw a dual-color fluorescence detection curve in real time according to the obtained dual-color fluorescence output value, which is convenient for the user to observe the detection result in real time.

[0049] Furthermore, in order to enable the color sensor to more accurately transmit the fluorescence of the two dyes to achieve the purpose of separating the fluorescence of the two dyes and eliminate the cross-interference between the fluorescences of the two dyes on the filters, we can further improve the filters in the color sensor: make filters with a relatively narrow filter color bandwidth according to the selected fluorescent dyes.

[0050] For example, the first filter is configured to transmit the fluorescence of the first dye and not transmit the fluorescence of the second dye, where the transmission wavelength of the first filter is the maximum fluorescence emission wavelength of the first dye ±10 to 25 nm;

[0051] The second filter is configured to transmit the fluorescence of the second dye and not transmit the fluorescence of the first dye, where the transmission wavelength of the second filter is the maximum fluorescence emission wavelength of the second dye ±10 to 25 nm.

[0052] Illustratively, FAM fluorescent dye, HEX fluorescent dye, and ROX fluorescent dye are commonly used fluorescent dyes. The excitation light wavelength required for the FAM fluorescent dye is 280 to 500 nm (preferably the excitation light wavelength is 480 nm), and the maximum fluorescence emission wavelength of the FAM fluorescent dye is at 520 nm.

[0053] The excitation light wavelength required for the HEX fluorescent dye is 480 to 530 nm (preferably the excitation light wavelength is 480 nm); the maximum fluorescence emission wavelength of the HEX fluorescent dye (i.e., the emission peak, that is, the wavelength corresponding to the maximum emission fluorescence intensity is called the maximum emission wavelength λem) is at 556 nm.

[0054] The excitation light wavelength required for the ROX fluorescent dye is 480 to 580 nm (preferably the excitation light wavelength is 480 nm); the maximum fluorescence emission wavelength of the ROX fluorescent dye is at 605 nm.

[0055] Thus, first, we can select blue light or ultraviolet light as the single excitation light source. The wavelength range of the single excitation light source is 260 to 500 nm. Preferably, the wavelength range of the single excitation light source is 380 to 480 nm. More preferably, the wavelength of the single excitation light source is 480 nm. Preferably, blue light is selected as the single excitation light source because we have tested and found that blue LED excitation light in this range can excite various fluorescent dyes, but the excitation efficiency is different.

[0056] Second, according to the selected fluorescent dyes, for example, if we select FAM fluorescent dye and ROX fluorescent dye as the dual fluorescent dyes, we can further narrow the wavelength range that the filter can transmit (i.e., narrow the filter color bandwidth). Since the maximum fluorescence emission wavelength of the FAM fluorescent dye is at 520 nm, we can set it as follows: the wavelength of the first filter is at 525 ± 25 nm. The maximum fluorescence emission wavelength of the ROX fluorescent dye is at 605 nm. Therefore, the wavelength of the corresponding second filter is at 600 ± 10 nm. Correspondingly, the wavelength of the filter corresponding to the HEX fluorescent dye can be 560 ± 10 nm.

[0057] By setting the filter in the color sensor to a filter with a relatively narrow color filtering bandwidth, the fluorescence of two fluorescent dyes can be transmitted and separated to the greatest extent and converted into electrical signals. After the control module processes the electrical signals, the dual-color fluorescence output value can be obtained more accurately. Further, in order to accurately transmit the fluorescence of the two fluorescent dyes, when selecting the fluorescent dyes, a group with a relatively large distance between the fluorescence emission wavelengths should be selected as much as possible. The greater the distance, the more accurate it is.

[0058] In this embodiment, the installation position of the single excitation light source is not limited, as long as it can excite the two fluorescent dyes in the reaction tube and the generated fluorescence emission light can be transmitted to the color sensor. Preferably, for example, the included angle between the excitation light rays generated by the single excitation light source and the fluorescence emission light rays reflected by the reaction tube is between 0 and 135°. Specifically, the single excitation light source 6 is arranged at the bottom of the reaction tube 8, and the included angle between the excitation light rays 61 generated by the single excitation light source and the fluorescence emission light rays 9 generated by the reaction tube is 90°, as shown in Figure 6. The single excitation light source 6 is arranged at the bottom of the reaction tube 8, and the included angle between the excitation light rays 61 generated by the single excitation light source and the fluorescence emission light rays 9 generated by the reaction tube 8 is 30°, as Figure 7 shown. The single excitation light source 6 is arranged at the top of the reaction tube 8, and the included angle between the excitation light rays 61 generated by the single excitation light source 6 and the fluorescence emission light rays 9 reflected by the reaction tube 8 is 0°, as Figure 8 shown. The single excitation light source 6 is arranged at the top of the reaction tube 8, and the included angle between the excitation light rays 61 generated by the single excitation light source 6 and the fluorescence emission light rays 9 generated by the reaction tube 8 is 90°, as Figure 9 shown.

[0059] In one embodiment, the color sensor can specifically be an RGB color sensor (RGB color sensor, RGB array sensor). The RGB color sensor has a filter array, and the filter array at least includes a first filter and a second filter. The first filter is used to transmit the G fluorescence of the first dye in the fluorescence emission light, and the second filter is used to transmit the R fluorescence of the second dye in the fluorescence emission light; the RGB sensor is configured to be able to transmit at least the G fluorescence and R fluorescence in the fluorescence emission light and convert the two fluorescences into electrical signals respectively, that is, the RGB sensor can convert the transmitted G fluorescence and R fluorescence into electrical signals respectively through the filter array it has; the control module collects the electrical signals in real time and processes them to obtain Δ the G, ΔR dual-color fluorescence output value.

[0060] Further, in order to obtain accurate Δ G, ΔR dual-color fluorescence output values, any one or a combination of the following two methods can be used to obtain accurate dual-color fluorescence output values.

[0061] The first method: We can also set the filter in the RGB sensor to be a filter with a relatively narrow color filter bandwidth made according to the selected fluorescent dye.

[0062] For example, the first filter is configured to transmit the fluorescence of the first dye, where the transmission wavelength of the first filter is the maximum fluorescence emission wavelength of the first dye ±10 - 25 nm;

[0063] The second filter is configured to transmit the fluorescence of the second dye, where the transmission wavelength of the second filter is the maximum fluorescence emission wavelength of the second dye ±10 - 25 nm.

[0064] For example, FAM fluorescent dye, HEX fluorescent dye, and ROX fluorescent dye are commonly used fluorescent dyes. The excitation light wavelength required for the FAM fluorescent dye is 280 - 500 nm (the preferred excitation light wavelength is 480 nm), and the maximum fluorescence emission wavelength of the FAM fluorescent dye is at 520 nm.

[0065] The excitation light wavelength required for the HEX fluorescent dye is 480 - 530 nm (the preferred excitation light wavelength is 480 nm); the maximum fluorescence emission wavelength of the HEX fluorescent dye is at 556 nm.

[0066] The excitation light wavelength required for the ROX fluorescent dye is 480 - 580 nm (the preferred excitation light wavelength is 480 nm); the maximum fluorescence emission wavelength of the ROX fluorescent dye is at 605 nm.

[0067] Therefore, we select blue light or ultraviolet light as the single excitation light source. The wavelength range of this single excitation light source is 260 - 500 nm. Preferably, the wavelength range of the single excitation light source is 380 - 480 nm. More preferably, the wavelength of the single excitation light source is 480 nm. It is preferred to select blue light as the single excitation light source because we have tested and found that using blue LED excitation light in this range can excite various fluorescent dyes, but the excitation efficiency is different.

[0068] Therefore, according to the selected fluorescent dye, for example, if we select FAM fluorescent dye and ROX fluorescent dye as the dual fluorescent dyes, we can further narrow the wavelength range that the filter can transmit (i.e., narrow the color filter bandwidth of the filter). Since the maximum fluorescence emission wavelength of the FAM fluorescent dye is at 520 nm, we can set it as: the wavelength of the first filter is at 525 ± 25 nm. And, the maximum fluorescence emission wavelength of the ROX fluorescent dye is at 605 nm. Therefore, the wavelength of the corresponding second filter is at 600 ± 10 nm. Correspondingly, the wavelength of the filter corresponding to the HEX fluorescent dye can be 560 ± 10 nm.

[0069] Therefore, we can set the filter in the RGB sensor to a filter with a narrow color filter bandwidth, which can transmit and separate the fluorescence of the two fluorescent dyes to the greatest extent, so that the fluorescence of the two dyes does not interfere with each other, and convert the fluorescence of the two dyes into electrical signals. After the control module processes the electrical signals, the dual-color fluorescence output value can be obtained more accurately.

[0070] Among them, the filter array of the RGB sensor is at least a 1 (filter with an R red fluorescence channel) * 1 (filter with a G color filter fluorescence channel) filter. Correspondingly, when we select fluorescent dyes, the fluorescent dyes should be selected to be able to produce fluorescence with a light wavelength of 390 - 780 nm.

[0071] For the first method above, corresponding filters need to be made according to the selected fluorescent dyes. However, this method has a high cost.

[0072] The second method: It can also be that, in order to save costs, we can also directly select common and commonly used RGB sensors on the market. As Figure 5 (a) shows, this RGB sensor is a square array composed of 9 optoelectronic devices and 9 filters (including but not limited to a 3×3 array, that is, R*3 / G*3 / B*3. The minimum is a 1×1×1 array, that is, R*1 / G*1 / B*1). By covering filters with different wavelength channels on the corresponding points of the optoelectronic device array, these filters form a filter array with different wavelength channels.

[0073] As Figure 5 (b), Figure 5 (c) shows, the filter array of a common and commonly used RGB sensor is composed of 3 filters with R red light channels, 3 filters with G green light channels, and 3 filters with B blue light channels. Each light channel corresponds to an optoelectronic device. Taking the filter for transmitting R red light as an example, as Figure 5 (b) shows, a filter for transmitting R red light (with an R red light channel) composed of a purple filter and a yellow filter stacked is provided on this optoelectronic device; it can also be that, as Figure 5 (c), only one red filter is provided on an optoelectronic device for transmitting R red light. Thus, the filter at each array point can be a single filter or composed of multiple filter slices stacked, as long as the filter finally constructed at the array point can transmit R red light, G green light, or B blue light, and there is no limit here.

[0074] If the filter is an array composed of R, G, and B color filters with a wide bandwidth, combined with the array composed of optoelectronic devices to form an RGB sensor, which is also a common RGB sensor on the market currently. To collect fluorescence data, intelligent algorithms need to be used for processing to accurately obtain the dual-channel fluorescence value.

[0075] This is because during the R & D process, we found that due to the detection wavelength width of each R, G, B array point of the existing low-cost RGB sensors and the detection wavelength overlap between the array points, the sensing of green, red, and blue fluorescence interferes with each other. This interference makes it difficult for the existing low-cost RGB sensors to be applied to fluorescence multiplex detection. For this reason, we assisted in developing an intelligent algorithm to process the acquired data, so as to obtain the fluorescence values corresponding to different wavelengths that can clearly distinguish two target analytes.

[0076] Among them, the RGB color sensor has a filter array, and the filter array includes at least a first filter and a second filter. The first filter is used to transmit the G fluorescence of two dyes in the fluorescence emission light, and the second filter is used to transmit the R fluorescence of two dyes in the fluorescence emission light; the RGB sensor converts the transmitted G fluorescence and R fluorescence into electrical signals through the filter array it has; the control module collects the electrical signals in real time and processes them to obtain the ΔG, ΔR dual-color fluorescence output values.

[0077] Since there is an overlap in the fluorescence wavelengths generated after the two dyes are excited, even if we select two dyes with a large distance between their fluorescence emission wavelengths, there will still be an overlap in the fluorescence wavelengths generated after they are excited. Therefore, in the filter array of the RGB sensor, the first filter for the G fluorescence channel transmits not only the G fluorescence emitted by the first dye but also the G fluorescence emitted by the second dye. Therefore, the first filter transmits the superimposed G fluorescence of the two dyes in the fluorescence emission light. The second filter transmits not only the R fluorescence emitted by the second dye but also the R fluorescence emitted by the first dye. Therefore, the second filter transmits the superimposed R fluorescence of the two dyes in the fluorescence emission light.

[0078] Therefore, in order to accurately obtain the dual-color fluorescence output values, we improved the processing method of the control module to eliminate the cross-interference between the fluorescences of the two dyes, so as to accurately obtain the fluorescence values of each fluorescent dye, and then obtain the concentrations of the two target analytes. Specifically, the control module obtains the ΔG, ΔR dual-color fluorescence output values through the following processing method: including,

[0079] Step 1): Convert the electrical signals collected in real time into R values and G values to obtain ΔR and ΔG, and then determine whether both ΔR and ΔG are > 0. If so, go to Step 2); if not, go to Step 3);

[0080] Step 2): When ΔR / ΔG < a, then the output value of this round ΔR k = ΔR k-1 , ΔG k = ΔG k-1 + ΔG;

[0081] When ΔR / ΔG > b, the output value of this round is ΔR k = ΔR k-1 + ΔR, ΔG k = ΔG k-1 ;

[0082] When a ≤ ΔR / ΔG ≤ b, the output value of this round is ΔR k = ΔR k-1 + ΔR, ΔG k = ΔG k-1 + ΔG;

[0083] Step 3): When ΔR or ΔG < 0, the output value of this round is ΔR k = ΔR k-1 , ΔG k = ΔG k-1 ;

[0084] where ΔR is the difference between the fluorescence values collected in this round and the previous round, and ΔG is the difference between the fluorescence values collected in this round and the previous round;

[0085] ΔR k and ΔG k are the output values of this round, and ΔR k-1 and ΔG k-1 are the output values of the previous round;

[0086] The value of a is the threshold of ΔR / ΔG that can completely distinguish the emission of the first dye and the non - emission of the second dye when the first dye emits light;

[0087] The value of b is the threshold of ΔR / ΔG that can completely distinguish the emission of the second dye and the non - emission of the first dye when the second dye emits light; and the maximum fluorescence emission wavelength of the first dye is less than the maximum fluorescence emission wavelength of the second dye.

[0088] Among them, when the two fluorescent dyes are determined, a and b are also determined. This is because the fluorescence wavelength range generated by one dye is relatively wide. When transmitted through the filter array in the RGB sensor, the fluorescence generated by one dye contains not only the transmissible R fluorescence but also the transmissible G fluorescence. Therefore, in order to determine the values of a and b respectively, we pre - adopt the single - tube single - color method, that is, only one dye is added to a single tube, and then through multiple statistical experiments. For example, when the first dye is excited to generate fluorescence, it is simultaneously transmitted through the first filter with a G fluorescence channel and the second filter with an R fluorescence channel in the RGB sensor, obtaining the transmitted G fluorescence value and R fluorescence value, thereby determining the ratio value of ΔR / ΔG corresponding to the first dye.

[0089] The second dye also uses the same method to obtain the R fluorescence value and G fluorescence value, and then determines the ratio value of ΔR / ΔG corresponding to the second dye.

[0090] Then, determine the threshold value of ΔR / ΔG when the first dye emits light, which can completely distinguish the emission of the first dye and the non-emission of the second dye, that is, the value of a. Determine the threshold value of ΔR / ΔG when the second dye emits light, which can completely distinguish the emission of the second dye and the non-emission of the first dye, that is, the value of b.

[0091] Therefore, in this application, by using an RGB array sensor to collect data of multiple points in a small range, and by obtaining these detected signal values and cooperating with the processing of the control module, the fluorescence values of different wavelengths corresponding to two target detection substances can be clearly confirmed, so that the dual-color fluorescence output value and the concentration content of the two target detection substances can be accurately obtained.

[0092] For example, the control module can also be connected to a terminal, and the terminal can draw a dual-color fluorescence detection curve in real time according to the dual-color fluorescence output value. The terminal can be a user terminal, such as a computer, a mobile phone, a pad, etc.

[0093] Of course, we can also combine the processing method of the control module with the color sensor having a filter with a narrow filter bandwidth, which can further improve the accuracy of dual-color fluorescence detection.

[0094] In one embodiment, the dual-color fluorescence detection component of this application further includes a high-pass filter. The high-pass filter is arranged in front of the color sensor and is used to first filter the excitation light in the fluorescence emission light, and the filtered fluorescence emission light is transmitted to the color sensor. The number of the color sensor, the high-pass filter, and the reaction tube is the same, and the three are arranged in one-to-one correspondence. Further, the structure of the high-pass filter is a convex lens structure, which is used to first filter the excitation light in the fluorescence emission light, and the filtered fluorescence emission light is focused and then transmitted to the color sensor.

[0095] Of course, between the reaction tube and the color sensor, a high-pass filter may not be included, but the color sensor filter array is used to directly filter the excitation light in the fluorescence emission light reflected by the reaction tube and transmit the filtered fluorescence emission light to the photosensitive layer of the color sensor.

[0096] That is, a high-pass filter may not be provided between the reaction tube and the color sensor. Instead, the filter array inherent in the color sensor is directly used to filter the excitation light in the fluorescence emission light reflected by the reaction tube while transmitting the fluorescence of the two dyes in the fluorescence emission light, and convert the two fluorescences into electrical signals respectively. For example, when the color sensor is an RGB color sensor, the RGB color sensor has a filter array, and the filter array includes at least a first filter and a second filter. The first filter is used to filter the excitation light in the fluorescence emission light while transmitting the G fluorescence of the first dye in the fluorescence emission light, and the second filter is used to filter the excitation light in the fluorescence emission light while transmitting the R fluorescence of the second dye in the fluorescence emission light; the RGB sensor converts the transmitted G fluorescence and R fluorescence into electrical signals respectively through the filter array it has; the control module collects the electrical signals in real time and processes them to obtain Δ G, ΔR dual-color fluorescence output values.

[0097] Further, for example, when the color sensor is an RGB color sensor, the RGB color sensor has a filter array, and the filter array includes at least a first filter and a second filter. The first filter is used to filter the excitation light in the fluorescence emission light while transmitting the G fluorescence of the two dyes in the fluorescence emission light, and the second filter is used to filter the excitation light in the fluorescence emission light while transmitting the R fluorescence of the two dyes in the fluorescence emission light; the RGB sensor converts the transmitted G fluorescence and R fluorescence into electrical signals respectively through the filter array it has; the control module collects the electrical signals in real time and processes them to obtain Δ G, ΔR dual-color fluorescence output values.

[0098] In a second aspect, according to an embodiment of the present disclosure, the present application further provides a POCT molecular diagnostic device, including a reaction tube and the dual-color fluorescence detection component in any one of the above embodiments. The POCT molecular diagnostic device with the dual-color fluorescence detection component can also achieve that the POCT molecular diagnostic device of the present application does not require a complex optical path design. Only a single excitation light source and a color sensor need to be set to complete the dual-color fluorescence detection. The structure is simple, the cost is low, it can be integrated into a micro device, and it is easy to carry. In the POCT molecular diagnostic device of the present application, by using an RGB array sensor to collect data of multiple points in a small range and cooperating with the processing method of the control module, the fluorescence values of different wavelengths corresponding to the two target analytes can be clearly confirmed, so that the dual-color fluorescence output values and the concentration contents of the two target analytes can be accurately obtained.

[0099] Both the reaction tube and the color sensor are provided with a plurality of them, and they are arranged at corresponding positions.

[0100] Further, the POCT molecular diagnostic device of the present application further includes a plurality of high-pass filters, which are arranged between the reaction tube and the color sensor, and are used to filter the excitation light in the fluorescence emission light reflected by the reaction tube and transmit the filtered fluorescence emission light to the color sensor.

[0101] The following further elaborates on the present application with specific application examples:

[0102] Example 1

[0103] As Figures 1 - 4 shown, a POCT molecular diagnostic device includes a carrier module 1, an RGB array sensor 2, an RGB array sensor mounting substrate 3, a filter mounting substrate 4, a high-pass filter 5, a single excitation light source 6, and a control module 7. The carrier module 1 is used to accommodate the reaction tube 8, and a first observation hole 11 is provided on the carrier module. The control module 7 is arranged inside the housing, and the single excitation light source 6 is connected to the control module 7 and is controlled by the control module 7 to be turned on or off. The single excitation light source 6 is arranged at the bottom of the reaction tube 8 to simultaneously excite two dyes in the reaction tube 8 to generate fluorescence excitation light. A plurality of high-pass filters 5 are provided and are embedded in the filter mounting substrate 4. The filter mounting substrate 4 is mounted on one side of the carrier module, and the high-pass filter 5 is arranged corresponding to the first observation hole 11, and is used to filter the excitation light in the fluorescence emission light reflected by the reaction tube and transmit the filtered fluorescence emission light to the RGB array sensor 2.

[0104] A plurality of RGB array sensors 2 are provided and a plurality of RGB array sensors 2 are all mounted on the RGB array sensor mounting substrate 3. The RGB array sensor mounting substrate 3 is mounted outside the filter mounting substrate 4, and the positions of the RGB array sensors 2 thereon are arranged corresponding to the high-pass filters 5 one by one. The RGB array sensor 2 is used to transmit the R fluorescence and G fluorescence of the two dyes in the fluorescence emission light and convert the R fluorescence and G fluorescence into electrical signals respectively. Specifically, the RGB color sensor has a filter array, and the filter array at least includes a first filter and a second filter. The first filter is used to transmit the G fluorescence of the two dyes in the fluorescence emission light, and the second filter is used to transmit the R fluorescence of the two dyes in the fluorescence emission light; the RGB sensor converts the transmitted G fluorescence and R fluorescence into electrical signals respectively through the filter array it has. The control module 7 is connected to the RGB array sensor 2 to receive the electrical signals in real time and convert them into R fluorescence values and G fluorescence values, and then obtain the ΔG and ΔR dual-color fluorescence output values through processing.

[0105] The control module processes the R fluorescence value and the G fluorescence value as follows, including:

[0106] Step 1): Convert the real-time collected electrical signal into R value and G value to obtain ΔR and ΔG. First, determine whether both ΔR and ΔG are > 0. If so, go to Step 2); if not, go to Step 3);

[0107] Step 2): When ΔR / ΔG < a, the output value of this round ΔR k = ΔR k-1 , ΔG k = ΔG k-1 + ΔG;

[0108] When ΔR / ΔG > b, the output value of this round ΔR k = ΔR k-1 + ΔR, ΔG k = ΔG k-1 ;

[0109] When a ≤ ΔR / ΔG ≤ b, the output value of this round ΔR k = ΔR k-1 + ΔR, ΔG k = ΔG k-1 + ΔG;

[0110] Step 3): When ΔR or ΔG < 0, the output value of this round ΔR k = ΔR k-1 , ΔG k = ΔG k-1 ;

[0111] Among them, ΔR is the difference between the fluorescence values collected in this round and the previous round, and ΔG is the difference between the fluorescence values collected in this round and the previous round;

[0112] ΔR k-1 and ΔG k-1 are the output values of the previous round;

[0113] The a value is the threshold of ΔR / ΔG that can completely distinguish the emission of the first dye and the non-emission of the second dye when the first dye emits light;

[0114] The b value is the threshold of ΔR / ΔG that can completely distinguish the emission of the second dye and the non-emission of the first dye when the second dye emits light; and the maximum fluorescence emission wavelength of the first dye is less than the maximum fluorescence emission wavelength of the second dye. The control module can also be connected to the terminal, and the terminal can draw a dual-color fluorescence detection curve in real time according to the output value, and then obtain the concentration content of the target analyte.

[0115] Among them, further, the structure of the high-pass filter 5 can also be a convex lens structure, which is used to first filter the excitation light in the fluorescence emission light and focus the filtered fluorescence emission light and transmit it to the RGB array sensor.

[0116] Example 2

[0117] As Figures 10 - 12 shown, Embodiment 2 provides a POCT molecular diagnostic device with another structure. This device also includes a carrier module 1, an RGB array sensor 2, an RGB array sensor mounting substrate 3, a filter mounting substrate 4, a high-pass filter 5, a single excitation light source 6, and a control module 7. The structural connection relationship, functional relationship among them are generally the same as those in Embodiment 1, and will not be elaborated here. The difference between Embodiment 2 and Embodiment 1 is that the number of reaction tubes 8 is eight, and correspondingly, 8 RGB array sensors are also provided. The structure of the high-pass filter 5 is a convex lens structure, which is used to first filter the excitation light in the fluorescence emission light and transmit the filtered and focused fluorescence emission light to the RGB array sensor 2.

[0118] It should be understood that various forms of the processes shown above can be used, reordering, adding, or deleting steps. For example, the steps described in this disclosure can be executed in parallel, sequentially, or in a different order, as long as the desired results of the technical solutions disclosed in this disclosure can be achieved, and no limitations are imposed herein.

[0119] In addition, the terms "first" and "second" are only used for descriptive purposes and cannot be construed as indicating or implying relative importance or implicitly specifying the quantity of the indicated technical features. Thus, the features defined with "first" and "second" may explicitly or implicitly include at least one of such features. In the description of this disclosure, "a plurality" means two or more unless otherwise specifically defined.

[0120] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples", etc. means that the specific features, structures, materials, or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of this disclosure. Moreover, the specific features, structures, materials, or characteristics described can be combined in a suitable manner in any one or more embodiments or examples.

[0121] In addition, without contradiction, those skilled in the art can combine and combine the different embodiments or examples described in this specification and the features of different embodiments or examples.

[0122] The above is only the specific implementation manner of this disclosure, but the protection scope of this disclosure is not limited thereto. Any person skilled in the art can easily think of changes or substitutions within the technical scope disclosed by this disclosure, and all should be covered by the protection scope of this disclosure. Therefore, the protection scope of this disclosure should be subject to the protection scope of the claimed rights.

Claims

1. A two-color fluorescence detection component, characterized in that: Comprising, A single excitation light source having a wavelength range, which is configured to be able to excite two dyes in a reaction tube to generate fluorescence emission light having different wavelength ranges; A color sensor, which is configured to be able to transmit the fluorescence of the two dyes in the fluorescence emission light and convert the two fluorescences into electrical signals respectively; A control module, connected to the color sensor, the control module is configured to process the electrical signals to obtain a dual-color fluorescence output value.

2. The dual-color fluorescence detection component according to claim 1, wherein: The color sensor has a filter array, the filter array at least includes a first filter and a second filter and the two have different wavelength channels, the first filter is configured to be able to transmit the fluorescence of the first dye in the fluorescence emission light and not transmit the fluorescence of the second dye, and the second filter is configured to be able to transmit the fluorescence of the second dye in the fluorescence emission light and not transmit the fluorescence of the first dye.

3. The dual-color fluorescence detection component according to claim 2, wherein: The transmission wavelength of the first filter is the maximum fluorescence emission wavelength of the first dye ±10 to 25 nm; The transmission wavelength of the second filter is the maximum fluorescence emission wavelength of the second dye ±10 to 25 nm.

4. The dual-color fluorescence detection component according to claim 1, wherein: The color sensor is an RGB color sensor. The RGB color sensor has a filter array. The filter array at least includes a first filter and a second filter. The first filter is used to transmit the G fluorescence of the first dye in the fluorescence emission light, and the second filter is used to transmit the R fluorescence of the second dye in the fluorescence emission light. The RGB color sensor converts the transmitted G fluorescence and R fluorescence into electrical signals through the filter array it has. The control module collects the electrical signals in real time and processes them to obtain Δ the G and ΔR dual-color fluorescence output values.

5. The dual-color fluorescence detection component according to claim 1, characterized in that: The color sensor is an RGB color sensor. The RGB color sensor has a filter array. The filter array at least includes a first filter and a second filter. The first filter is used to transmit the G fluorescence of two dyes in the fluorescence emission light, and the second filter is used to transmit the R fluorescence of two dyes in the fluorescence emission light. The RGB color sensor converts the transmitted G fluorescence and R fluorescence into electrical signals respectively through the filter array it has. The control module collects the electrical signals in real time and processes them to obtain Δ G, Δ R bicolor fluorescence output values.

6. The dual-color fluorescence detection component according to claim 5, characterized in that: When the color sensor is an RGB color sensor, the fluorescent dyes used are configured to be able to generate fluorescence with a light wavelength range of 390 to 780 nm.

7. The dual-color fluorescence detection component according to any one of claims 1-6, characterized in that: The single excitation light source uses blue light or ultraviolet light, and the wavelength range of the single excitation light source is 260 to 500 nm.

8. The dual-color fluorescence detection component according to claim 7, wherein: The wavelength range of the single excitation light source is 380 to 480 nm.

9. The dual-color fluorescence detection component according to any one of claims 1-6, characterized in that: Further comprising a high-pass filter, the high-pass filter is arranged in front of the color sensor, and is used to first filter the excitation light in the fluorescence emission light and transmit the filtered fluorescence emission light to the color sensor; The color sensor, the high-pass filter, and the reaction tube are arranged in one-to-one correspondence.

10. The dual-color fluorescence detection component according to any one of claims 1-6, characterized in that: The color sensor is configured to be able to filter the excitation light in the fluorescence emission light while transmitting the fluorescence of the two dyes in the fluorescence emission light and converting the two fluorescences into electrical signals respectively.

11. A POCT molecular diagnostic device, characterized in that: Comprising the dual-color fluorescence detection component according to any one of claims 1-10.

Images