Information recognition method and hardware system of a fluorescence detection device

Abstract

This application provides an information identification method and hardware system for a fluorescence detection device, relating to the field of medical testing technology. The method includes: acquiring a reaction tube position coding scheme for a preset biological detection item using the fluorescence detection device; performing fluorescence detection on the well plate of the fluorescence detection device during the fluorescence detection process of the preset biological detection item to obtain channel fluorescence values ​​for multiple well positions on the well plate; determining the preset channel combination information of the target reaction tube position corresponding to the well position from preset channel combinations of multiple reaction tube positions based on the channel fluorescence values ​​of each well position; and determining the sub-detection item carried by the well position under the preset biological detection item based on the preset channel combination information. This application improves the fault tolerance, automation level, and data traceability of the detection process.

Description

Technical Field

[0001] This application relates to the field of medical testing technology, and more specifically, to an information identification method and hardware system for a fluorescence detection device. Background Technology

[0002] In clinical laboratory and molecular diagnostics practice, simultaneous screening for multiple pathogens has become a rigid requirement in scenarios such as respiratory infections, multidrug resistance monitoring, and tumor multigenotyping. To improve throughput, researchers commonly use high-density microplates to run multiple independent multiplex fluorescent polymerase chain reaction (PCR) tubes in parallel. Each reaction tube carries different combinations of target detection items, and relies on the multiple optical channels of the PCR instrument to identify the corresponding fluorescently labeled probes.

[0003] In the prior art, before the multiplex fluorescent polymerase chain reaction detection experiment, the operator needs to set the corresponding reagent tube position number, detection target and fluorescence channel in the instrument software for each well. During the operation, the instrument only records the fluorescence signal according to the preset position and does not verify whether the actual content loaded at that position is consistent with the preset.

[0004] However, in existing technologies, tube misplacement or sequence confusion may occur during the sample addition process. The instrument cannot detect the deviation, resulting in a complete mismatch between the fluorescence signal and the preset items, leading to irreversible misjudgment. Furthermore, it is impossible to deduce the true tube location from the signal pattern after the detection is completed. When abnormal results occur, it is necessary to manually compare the physical well positions, sample addition records, preset configurations, and original fluorescence curves one by one. The traceability process is cumbersome and inefficient, which seriously restricts the reliability and automation level of high-throughput detection. Summary of the Invention

[0005] The purpose of this application is to address the shortcomings of the prior art by providing an information identification method and hardware system for a fluorescence detection device, thereby improving the fault tolerance, automation, and data traceability of the detection process.

[0006] To achieve the above objectives, the technical solutions adopted in the embodiments of this application are as follows: In a first aspect, one embodiment of this application provides an information identification method for a fluorescence detection device, the method comprising: The reaction tube position coding scheme of the fluorescence detection device for the preset biological detection project is obtained. The reaction tube position coding scheme includes: preset channel combination information of multiple reaction tube positions; the preset channel combination information of each reaction tube position includes: the activation detection channel corresponding to the reaction tube position and the target information of each activation detection channel; the preset channel combination information of different reaction tube positions is different. During the fluorescence detection process of the preset biological detection item, fluorescence detection is performed on the well plate of the fluorescence detection device to obtain the channel fluorescence values ​​of multiple well positions on the well plate; the channel fluorescence value of each well position includes: the fluorescence value of the detection channel corresponding to the well position; Based on the channel fluorescence value of each well, the preset channel combination information of the target reaction tube corresponding to the well is determined from the preset channel combinations of the multiple reaction tube positions; Based on the preset channel combination information, determine the sub-detection items carried by the pore position under the preset biological detection items.

[0007] Optionally, before acquiring the reaction tube location coding scheme for the preset biological detection item using the fluorescence detection device, the method further includes: Channel settings are performed on multiple preset reaction tube positions to obtain channel combination information for multiple preset reaction tube positions. The channel combination information for each preset reaction tube position includes activation indication information for at least one detection channel. Based on the information of multiple targets to be tested corresponding to the preset biological detection items, and the channel combination information of multiple preset reaction tube positions, the reaction tube position coding scheme is determined.

[0008] Optionally, the step of setting channels for multiple preset reaction tube positions to obtain channel combination information for the multiple preset reaction tube positions includes: Based on preset non-zero constraint conditions and uniqueness constraint conditions, channel settings are performed on multiple preset reaction tube positions to obtain channel combination information of multiple preset reaction tube positions; the preset non-zero constraint conditions are used to indicate that each preset reaction tube position activates at least one detection channel, and the uniqueness constraint conditions are used to indicate that the channel combination information of different preset reaction tube positions is different.

[0009] Optionally, the target information includes: the total number of targets and the type of each target; determining the reaction tube location coding scheme based on the target information corresponding to the preset biological detection item and the channel combination information of multiple preset reaction tube locations includes: Obtain the Hamming distance between every two channel combinations in the channel combination information of multiple preset reaction tube positions, and determine the minimum Hamming distance; Based on the total number of targets, the required number of preset reaction tube positions, and the type of each target, the set of channel combinations with the largest minimum Hamming distance is selected from the channel combination information of multiple preset reaction tube positions as the reaction tube position coding scheme, using the preset target total number constraint condition. The preset target total number constraint condition is used to indicate that the number of activation detection channels for all reaction positions in the reaction position coding scheme is equal to the total number of targets.

[0010] Optionally, determining the preset channel combination information of the target reaction tube position corresponding to the well position from the preset channel combination information of the plurality of reaction tube positions based on the channel fluorescence value of each well position includes: Based on the channel fluorescence value of each pore, calculate the fluorescence channel characteristics of each pore. Based on the fluorescence channel characteristics of each well site and the preset channel combination information of each reaction tube site, the channel feature similarity between the well site and each reaction tube site is calculated respectively. Based on the channel feature similarity, the reaction tube position corresponding to the highest similarity is determined as the target reaction tube position corresponding to the orifice position, and the preset channel combination information of the target reaction tube position is determined.

[0011] Optionally, calculating the fluorescence channel characteristics of each well site based on the channel fluorescence value of each well site includes: Obtain the preset fluorescence threshold; Based on the preset fluorescence threshold, the channel fluorescence value of each pore is processed to obtain the fluorescence channel characteristics of each pore.

[0012] Optionally, obtaining the preset fluorescence threshold includes: Obtain the instrument background value of each well and the standard deviation of the original fluorescence value of each well. Based on the instrument background value of each well in the same detection channel and the standard deviation of the original fluorescence value of the channel, a first channel preset fluorescence threshold is determined for each detection channel. The standard deviation of the original fluorescence value of the channel is the standard deviation of the original fluorescence value of the channel when no fluorescent probe is placed. The preset fluorescence threshold includes the first channel fluorescence threshold of each detection channel. Alternatively, based on the original fluorescence value of each well in the same detection channel, the preset coefficient, and the instrument background value of each well, a second channel preset fluorescence threshold is determined for each detection channel, wherein the preset fluorescence threshold includes the second channel fluorescence threshold for each detection channel.

[0013] Optionally, determining the reaction tube position corresponding to the highest similarity based on the channel feature similarity as the target reaction tube position corresponding to the aperture position includes: Based on the channel feature similarity of all the said orifice locations and each of the said reaction tube locations, a feature vector matching matrix is ​​constructed; The matching matrix is ​​assigned values ​​using a preset assignment method to obtain a new feature vector matching matrix. Based on the new feature vector matching matrix, the reaction tube position corresponding to the highest similarity is determined as the target reaction tube position corresponding to the well position.

[0014] Optionally, the step of performing fluorescence detection on the well plate of the fluorescence detection device to obtain the channel fluorescence values ​​of multiple well positions on the well plate includes: The fluorescence detection device is subjected to multiple cycles of fluorescence detection on the well plate to obtain the initial channel fluorescence values ​​of multiple well positions in multiple cycles; The channel fluorescence values ​​of multiple well sites on the plate are determined based on the initial channel fluorescence values ​​of multiple well sites in multiple cycles.

[0015] Secondly, another embodiment of this application provides an information identification device for a fluorescence detection device, the device comprising: The acquisition module is used to acquire the reaction tube position coding scheme of the fluorescence detection device for a preset biological detection item. The reaction tube position coding scheme includes: preset channel combination information of multiple reaction tube positions; the preset channel combination information of each reaction tube position includes: the activation detection channel corresponding to the reaction tube position and the target information of each activation detection channel. The detection module is used to perform fluorescence detection on the well plate of the fluorescence detection device during the fluorescence detection process of the preset biological detection item, and obtain the channel fluorescence values ​​of multiple well positions on the well plate; the channel fluorescence value of each well position includes: the fluorescence value of the detection channel corresponding to the well position; The determining module 1 is used to determine the preset channel combination information of the target reaction tube position corresponding to the well position from the preset channel combinations of the multiple reaction tube positions based on the channel fluorescence value of each well position; The determining module 2 is used to determine the sub-detection items carried by the well position under the preset biological detection items based on the preset channel combination information.

[0016] Thirdly, another embodiment of this application provides a hardware system including: a processor, a memory, and a bus. The memory stores machine-readable instructions executable by the processor. When the hardware system is running, the processor communicates with the memory via the bus, and the processor executes the machine-readable instructions to perform the steps of the information recognition method of the fluorescence detection device as described in any of the first aspects above.

[0017] Fourthly, another embodiment of this application provides a storage medium storing a computer program, which, when executed by a processor, performs the steps of the information identification method of any of the fluorescence detection devices described in the first aspect above.

[0018] The beneficial effects of this application are: This application provides an information identification method and hardware system for a fluorescence detection device. It acquires a coding scheme for the reaction tube positions of a fluorescence detection device for a preset biological detection item. During the fluorescence detection process of the preset biological detection item, fluorescence detection is performed on the well plate of the fluorescence detection device to obtain the channel fluorescence values ​​of multiple well positions on the well plate. Based on the channel fluorescence values ​​of each well position, the preset channel combination information of the target reaction tube position corresponding to the well position is determined from the preset channel combinations of multiple reaction tube positions. Based on the preset channel combination information, the sub-detection item carried by the well position under the preset biological detection item is determined. This application avoids reliance on manually preset tube position information and physical markings. By encoding the tube position identification information into the fluorescence channel combination of the reagent itself, the fluorescence value of the well position collected by the fluorescence detection device is automatically matched and identified to identify the reaction tube position it carries. This achieves intelligent association between the detection content and the physical location, improving the fault tolerance, automation level, and data traceability of the detection process. Furthermore, it requires no modification to existing instrument hardware and has good compatibility and practicality. Attached Figure Description

[0019] To more clearly illustrate the technical solutions of the embodiments of this application, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of this application and should not be regarded as a limitation of the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0020] Figure 1 A schematic flowchart illustrating an information identification method for a fluorescence detection device provided in an embodiment of this application; Figure 2 A flowchart illustrating the process of determining the reaction tube location coding scheme in an information identification method for a fluorescence detection device provided in this application embodiment; Figure 3 A flowchart illustrating the determination of channel combination information in another information identification method for a fluorescence detection device provided in this application embodiment; Figure 4 A schematic flowchart illustrating the process of determining the target reaction tube position in an information identification method for a fluorescence detection device provided in this application embodiment; Figure 5 A flowchart illustrating the process of determining fluorescence channel features in an information identification method for a fluorescence detection device provided in this application embodiment; Figure 6 A schematic diagram illustrating the process of determining a preset fluorescence threshold in an information identification method for a fluorescence detection device provided in an embodiment of this application; Figure 7 A schematic flowchart illustrating the process of determining the target reaction tube position in another information identification method for a fluorescence detection device provided in this application embodiment; Figure 8 This is a flowchart illustrating the process of determining channel fluorescence values ​​in an information identification method for a fluorescence detection device provided in an embodiment of this application. Figure 9 A cycle-based original fluorescence curve provided for an embodiment of this application; Figure 10 A schematic diagram of an amplification curve provided in an embodiment of this application; Figure 11 Another cycle-based original fluorescence curve provided for embodiments of this application; Figure 12 This is another schematic diagram of an amplification curve provided in an embodiment of this application; Figure 13 This is a schematic diagram of the structure of an information recognition device for a fluorescence detection device provided in an embodiment of this application; Figure 14 This is a schematic diagram of a hardware system provided in an embodiment of this application. Detailed Implementation

[0021] To make the objectives, technical solutions, and advantages of the embodiments of this application clearer, the technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. It should be understood that the accompanying drawings in this application are for illustrative and descriptive purposes only and are not intended to limit the scope of protection of this application. Furthermore, it should be understood that the schematic drawings are not drawn to scale. The flowcharts used in this application illustrate operations implemented according to some embodiments of this application. It should be understood that the operations in the flowcharts may not be implemented in sequence, and steps without logical contextual relationships may be reversed or implemented simultaneously. In addition, those skilled in the art, guided by the content of this application, may add one or more other operations to the flowcharts, or remove one or more operations from the flowcharts.

[0022] Furthermore, the described embodiments are merely some, not all, of the embodiments of this application. The components of the embodiments of this application described and illustrated herein can typically be arranged and designed in various different configurations. Therefore, the following detailed description of the embodiments of this application provided in the accompanying drawings is not intended to limit the scope of the claimed application, but merely to illustrate selected embodiments of the application. All other embodiments obtained by those skilled in the art based on the embodiments of this application without inventive effort are within the scope of protection of this application.

[0023] It should be noted that the term "comprising" will be used in the embodiments of this application to indicate the presence of the features declared thereafter, but does not exclude the addition of other features.

[0024] The development of real-time fluorescence polymerase chain reaction (PCR) technology is rooted in breakthroughs in polymerase chain reaction PCR. With the increasing demand for multiplex detection, researchers developed multiplex fluorescence PCR technology. This technology uses multiple TaqMan probes labeled with different fluorophores in the same reaction tube. Each probe targets a different target gene, and each probe's reporter group has characteristic excitation and emission wavelengths, thus enabling the simultaneous detection of multiple targets in a single tube. With the emergence of the concept of multiplex fluorescence PCR, the requirement for multi-channel fluorescence detection devices has also arisen.

[0025] Current fluorescence detection devices typically feature 4 to 6 independent fluorescence channels, each containing excitation and emission filters of specific wavelengths to match the optical properties of the corresponding fluorophore. This multi-channel optical system allows for the simultaneous detection of up to 4 to 6 different target genes in the same reaction tube, significantly enhancing the information capacity of a single tube and meeting some of the needs for multiplex pathogen screening in clinical diagnosis. However, with the further expansion of detection throughput, the multiplex detection capability of a single tube is no longer sufficient to meet the growing detection demands, making parallel detection across multiple tube positions essential for multiplex amplification. In the multiplex fluorescence PCR multi-tube detection mode, researchers need to pre-set the detection items for each well of the multiplex fluorescence detection device using software before detection, including sample name, target type, fluorescence channel, etc. This limits the existing multiplex fluorescence PCR multi-tube detection technology to pre-set and execution modes, making it overly reliant on physical markers. If the sample is misplaced, the instrument cannot detect it, leading to incorrect result matching. Furthermore, the system cannot identify the tube content from the data itself, resulting in low data analysis efficiency.

[0026] Therefore, this application provides an information identification method for a fluorescence detection device. By using a preset tube position coding scheme, and based on the channel fluorescence values ​​of multiple well positions during fluorescence detection, the preset channel combination information corresponding to each well position is determined, thereby obtaining the sub-detection item carried by the well position. This application reduces the absolute reliance on physical location markings and avoids the time-consuming and laborious process of manual verification and traceability.

[0027] To clearly describe the information identification method of the fluorescence detection device provided in the embodiments of this application, the method will be described below in conjunction with several accompanying drawings. Figure 1 This is a flowchart illustrating an information recognition method for a fluorescence detection device provided in an embodiment of this application, as shown below. Figure 1 As shown, the method includes: Step 101: Obtain the reaction tube location coding scheme of the fluorescence detection device for the preset biological detection items.

[0028] The reaction tube coding scheme includes: preset channel combination information for multiple reaction tube positions. The preset channel combination information for each reaction tube position includes: the activation detection channel corresponding to the reaction tube position and the target information for each activation detection channel; the preset channel combination information differs for different reaction tube positions. The preset biological detection items constitute the entire testing protocol, indicating all the sub-items to be measured in this test. For example, it could be a ten-item test for human respiratory pathogens, simultaneously detecting ten sub-items.

[0029] Optionally, the fluorescence detection device has a well plate, with each well used to place a reaction tube. Different reaction tubes contain different combinations of detection items, and each detection item occupies one fluorescence channel. The number of fluorescence channels in each reaction tube is determined by the fluorescence detection device, which is a multiplex fluorescence PCR instrument. For example, if the fluorescence detection device has four fluorescence channels, then each reaction tube has four fluorescence channels. Generally, a fixed fluorescence channel is used for quality control (internal control, or IC) in the reaction tube. Therefore, one fluorescence channel in each reaction tube is used for quality control, and the other three fluorescence channels can detect up to three detection items simultaneously.

[0030] Each reaction tube position is a reagent identifier with a unique fluorescence channel code and is not used to indicate its physical location on the well plate. It is identified by its logical number. For example, tube A is for COVID-19 single-sample testing, and tube B is for both influenza A and influenza B detection. The preset channel combination information for multiple reaction tube positions corresponds to the preset channel combination information for the respective reaction tubes. For example, it could be the preset channel combination information for tubes A and B. Activating the detection channel indicates whether the fluorescence channel in the reaction tube position is activated. When fluorescence channel CH1 is activated, it can be recorded as 1; when fluorescence channel CH2 is not activated, it can be recorded as 0. For example, if there are four fluorescence channels in the fluorescence detection device, and fluorescence channel CH1 is activated, fluorescence channel CH2 is activated, fluorescence channel CH3 is not activated, and fluorescence channel CH4 is activated in tube A, the corresponding activation detection channel code for tube A would be [1, 1, 0, 1]. The target information for each activated detection channel differs across different reaction tube locations. This target information indicates the identity of the target substance detected by that fluorescence channel. In fluorescence detection devices, the target information can be a specific nucleic acid series, such as a specific gene of a virus. For example, the activated detection channel of tube A is coded as [1, 1, 0, 1]. The target information for fluorescence channel CH1 in tube A is influenza A virus (Flu A), the target information for fluorescence channel CH2 is influenza B virus (Flu B), and the target information for fluorescence channel CH4 is IC.

[0031] Optionally, based on the targets that need to be detected at each reaction tube position in the preset biological detection project, and combined with the available fluorescence channels of the fluorescence detection device, determine which channels are activated at each reaction tube position and the target information corresponding to each channel, and save this configuration table as a coding scheme.

[0032] Optionally, based on the target combination contained in each reaction tube position in the preset biological detection project, available channel allocation schemes are retrieved from the preset coding rule base to generate preset channel combination information for multiple reaction tube positions.

[0033] Step 102: During the fluorescence detection process of the preset biological detection items, fluorescence detection is performed on the well plate of the fluorescence detection device to obtain the channel fluorescence values ​​of multiple wells on the well plate.

[0034] The fluorescence value of each well includes the fluorescence value of the detection channel corresponding to that well. The fluorescence channel corresponding to each well is determined by the fluorescence detection device. When the fluorescence detection device has four fluorescence channels, the fluorescence value of the well is the fluorescence value of each well in all four fluorescence channels. The fluorescence value is the fluorescence intensity value of that fluorescence channel. The wells in the well plate of the fluorescence detection device are placed randomly and not according to the order in the reaction tube coding scheme.

[0035] Optionally, fluorescence detection is used to detect the fluorescence intensity of the fluorescence signal. The principle of fluorescence signal generation is that the 5' end of the TaqMan probe is labeled with a reporter fluorescent group, and the 3' end is labeled with a quencher group. When the probe is intact, the quencher group inhibits the fluorescence emission of the reporter group through fluorescence resonance energy transfer. However, the quenching efficiency is not 100%, and the probe molecule always has a weak residual fluorescence in the solution. Therefore, when there is no amplification, the fluorescence value of the channel containing the fluorescent probe is higher than that of the channel without the probe. The principle of fluorescence signal detection is that during the PCR annealing stage, the TaqMan probe specifically binds to the target DNA sequence. During the extension stage, TaqDNA polymerase synthesizes a new strand along the template strand. When it encounters the probe bound to the template, it uses the exonuclease activity from the 5' end to the 3' end of the TaqMan probe to cleave the probe one by one. After the probe is cleaved, the reporter group and the quencher group separate, the distance between them increases, and the quenching effect disappears. Under excitation light, the reporter group emits fluorescence at a characteristic wavelength, which is captured by the instrument detection system to obtain the fluorescence signal.

[0036] Optionally, during the fluorescence detection process of the preset biological detection items, the fluorescence detection device performs endpoint fluorescence reading or steady-state fluorescence scanning on the well plate. Each well is sequentially illuminated, and the emission intensity of each well under each fluorescence channel is collected through a filtering system of different wavelengths, thereby directly obtaining the channel fluorescence value corresponding to each channel for each well.

[0037] Step 103: Based on the channel fluorescence value of each well, determine the preset channel combination information of the target reaction tube corresponding to the well from the preset channel combinations of multiple reaction tubes.

[0038] The target reaction tube position is the reaction tube position type in the coding scheme corresponding to the orifice position.

[0039] Optionally, after obtaining the channel fluorescence value of each well, the activation detection channel of each well is determined based on the fluorescence value of each well under all detection channels. The activation detection channel of each well is then matched with the activation detection channels of multiple reaction tube positions in the reaction tube position coding scheme to obtain preset channel combination information corresponding to the target reaction tube position for each well.

[0040] Step 104: Based on the preset channel combination information, determine the sub-detection items carried by the well sites under the preset biological detection items.

[0041] The sub-test items are multiple test items within the preset biological test items. When the preset biological test items are a respiratory tract ten-in-one test, the sub-test items can be influenza A virus and influenza B virus.

[0042] Optionally, using the target reaction tube location as an index, the reaction tube location coding scheme table is consulted to determine the target information for each activated detection channel of the target reaction tube location. Based on the amplification curve of this well location under each fluorescence channel, the positive or negative result of each sub-detection item is determined. The names and positive / negative results of all sub-detection items under this well location are summarized and output to obtain the sub-detection items and their corresponding detection results.

[0043] This application provides an information identification method for a fluorescence detection device. The method involves obtaining a coding scheme for the reaction tube positions of the fluorescence detection device for a preset biological detection item. During the fluorescence detection process of the preset biological detection item, fluorescence detection is performed on the well plate of the fluorescence detection device to obtain the channel fluorescence values ​​of multiple well positions on the well plate. Based on the channel fluorescence values ​​of each well position, the preset channel combination information of the target reaction tube position corresponding to the well position is determined from the preset channel combinations of multiple reaction tube positions. Based on the preset channel combination information, the sub-detection item carried by the well position under the preset biological detection item is determined. This application avoids reliance on manually preset tube position information and physical markings. By encoding the tube position identification information into the fluorescence channel combination of the reagent itself, the fluorescence value of the well position collected by the fluorescence detection device is automatically matched and identified to identify the reaction tube position it carries. This achieves the association between the detection content and the physical location, improving the fault tolerance, automation, and data traceability of the detection process. Furthermore, it requires no modification to existing instrument hardware and has good compatibility and practicality.

[0044] Based on the above embodiments, this application provides a process for determining the reaction tube position coding scheme in the information identification method of a fluorescence detection device. Figure 2 This is a flowchart illustrating the process of determining the reaction tube location coding scheme in an information identification method for a fluorescence detection device provided in this application embodiment, as shown below. Figure 2 As shown, before obtaining the reaction tube location coding scheme for the preset biological detection item in step 101 above, the method further includes: Step 201: Set the channels for multiple preset reaction tube positions to obtain the channel combination information of multiple preset reaction tube positions.

[0045] The channel combination information for each preset reaction tube position includes activation indication information for at least one detection channel. Channel settings determine the location and number of activated channels in the fluorescence channels of each preset reaction tube position.

[0046] Optionally, based on the total number of detection targets required for the preset biological detection project and the number of available fluorescence channels, the number of reaction tube positions and the number of channels to be activated for each position are determined. Channel settings are configured for multiple preset reaction tube positions to obtain channel combination information for multiple preset reaction tube positions.

[0047] For example, for tube position j, its N-dimensional fluorescence channel preset row vector is defined as: in {0, 1} This indicates that the i-th fluorescence channel at tube position j is activated (including the fluorescent probe targeting a specific target). This indicates that the device is not activated (it does not contain a fluorescent probe).

[0048] Step 202: Determine the reaction tube coding scheme based on the information of multiple targets to be tested corresponding to the preset biological detection items and the channel combination information of multiple preset reaction tube positions.

[0049] Optionally, based on the total number of all targets to be tested included in the preset biological detection project and the number of available fluorescence channels in the channel combination information of the preset reaction tube position, each activated channel is bound to the target to be tested, thereby forming complete coding information for each reaction tube position, which serves as the reaction tube position coding scheme.

[0050] In this embodiment, the identification and detection functions of the reaction tube positions are organically integrated. By planning the channel activation state of each tube position during the reagent design stage, the combination of fluorescence channels in each tube position is made unique, thereby providing a reliable basis for subsequent automatic identification based on fluorescence signals. This avoids the error risk caused by manual settings and achieves a synergistic improvement in the consistency and identifiability of the detection system.

[0051] Based on the above embodiments, this application also provides a process for determining channel combination information in an information identification method for a fluorescence detection device. In step 201 above, channels are set for multiple preset reaction tube positions to obtain channel combination information for multiple preset reaction tube positions, including: Based on the preset non-zero constraint and uniqueness constraint, channel settings are performed on multiple preset reaction tube positions to obtain channel combination information for multiple preset reaction tube positions.

[0052] Among them, the preset non-zero constraint condition is used to indicate that each preset reaction tube position activates at least one detection channel, and the uniqueness constraint condition is used to indicate that the channel combination information of different preset reaction tube positions is different.

[0053] Optionally, based on the total number of channels in the fluorescence detection device, all non-zero channel activation vectors are determined according to a preset non-zero constraint. Then, based on a unique constraint, each preset reaction tube position is assigned a distinct activation vector to ensure that at least one channel in each tube position is activated and that the activation modes of any two tube positions are not completely identical, thereby obtaining channel combination information for multiple preset reaction tube positions.

[0054] In this embodiment, by introducing dual logical constraints of non-zeroness and uniqueness, it is ensured that each reaction tube position has an identifiable fluorescent characteristic basis, while ensuring that the channel combinations between each tube position are distinguishable from each other, avoiding the risk of identification failure caused by empty combinations, and eliminating the identification ambiguity problem caused by the same combination, thereby improving the robustness and determinism of the entire detection system.

[0055] Based on the above embodiments, the target information includes: the total number of targets and the type of each target. This application also provides a process for determining channel combination information in another information identification method for a fluorescence detection device. Figure 3 A flowchart illustrating the determination of channel combination information in another fluorescence detection device information identification method provided in this application embodiment is shown below. Figure 3 As shown, in step 202 above, based on the target information corresponding to the preset biological detection item and the channel combination information of multiple preset reaction tube positions, the reaction tube position coding scheme is determined, including: Step 301: Calculate the Hamming distance between every two channel combinations in the channel combination information of multiple preset reaction tube positions, and determine the minimum Hamming distance.

[0056] The Hamming distance measures the number of different characters at corresponding positions between two strings (or vectors) of equal length. For the channel activation feature vector of the activation detection channel corresponding to the reaction tube position, the Hamming distance is the number of dimensions with different values ​​in the two vectors.

[0057] Optionally, based on the channel activation feature vector of the activated detection channel in the channel combination information of multiple preset reaction tube positions, a preset calculation formula is used to determine the Hamming distance between the channel activation feature vectors of every two channel combinations in the channel combination information of multiple preset reaction tube positions, and the minimum Hamming distance is determined from the Hamming distance between the channel activation feature vectors of every two channel combinations.

[0058] Step 302: Based on the total number of targets, the required number of preset reaction tube positions, and the type of each target, and using the preset target total number constraint, select the channel combination set with the largest minimum Hamming distance from the channel combination information of multiple preset reaction tube positions as the reaction tube position coding scheme.

[0059] Among them, the preset target total number constraint condition is used to indicate that the number of activation detection channels of all reaction tube positions in the reaction tube position coding scheme is equal to the total number of targets.

[0060] Optionally, based on the preset number of reaction tube positions required, all candidate sets that satisfy the non-zero and uniqueness constraints are enumerated from the non-zero channel combinations. Then, for each candidate set, the Hamming distance between any two channel combination information is calculated, and the minimum Hamming distance of the set is found. Finally, the channel combination set with the largest minimum Hamming distance and simultaneously satisfying the total number of targets constraint is selected from all candidate sets as the final reaction tube position encoding scheme.

[0061] For example, a preset calculation formula is used. Calculate the minimum Hamming distance among the Hamming distances of the activation feature vectors of every two channels, where, A matrix composed of channel combination information for preset reaction tube positions. For matrix Minimum Hamming distance, Given any two rows from row 1 to row R, where j and k are different, To calculate how many positions in two binary vectors, row j and row k, have different values, Let j be the vector in row j. Let be the vector in the k-th row.

[0062] For example, assuming the fluorescence detection device has 4 channels, 3 effective fluorescence channels excluding the IC, a preset requirement of 5 reaction tube positions, and a total of 10 targets, select distinct non-zero binary vectors for the 5 tube positions in the effective 3D space (CH1–CH3), and satisfy the following conditions: .

[0063] Among them, the 3-dimensional binary space has a total of There are three non-zero vectors, with the following number of weights: weight 1 (100, 010, 001) has 3 weights, weight 2 (110, 101, 011) has 3 weights, and weight 3 (111) has 1 weight. This satisfies the equation... Furthermore, the unique weighting, consisting of five distinct vectors, is divided into {3, 2, 2, 2, 1}. This means that only one weighted vector of 3 (111), all three weighted vectors of 2 (110, 101, 011), and any one weighted vector of 1 (represented by 100, equivalent to 010 and 001) can be selected. The weight is the number of 1s in the vector, which represents the number of targets detected simultaneously at one reaction tube position.

[0064] For example, the fluorescence detection device has 3 effective channels (excluding IC), a preset requirement of 5 reaction tube positions, and a total of 10 targets. The reaction tube position coding scheme is shown in the following table: Reaction tube location coding scheme table

[0065] In a 3D fluorescence channel, what is the minimum Hamming distance among 5 selected from 7 non-zero vectors? The theoretical upper bound is 1 (there must exist a pair of neighbors with a Hamming distance of 1 among any 5 distinct 3D non-zero binary vectors, because 3D space cannot simultaneously maintain the distance between 5 vectors). ≥2). This design reaches this upper limit, that is... =1, constituting the optimal design under this constraint. Pipe 4 (011) and pipe 5 (100) are complementary vectors, and their 3D Hamming distance dH = 3 (maximum value) corresponds to the Pearson correlation coefficient r( )= 1. Achieve perfect negative correlation. Pipe 1 corresponds to a 3-dimensional vector of all 1s. =(1,1,1), with a mean of 1 and a variance of 0, and the Pearson correlation coefficient is undefined. When the measured eigenvector... At that time, the preset all-1 pipe position can be directly searched for precise matching without calculating the correlation coefficient.

[0066] Separability of tubes 2-5: For tubes 2-5 (CH1-3 channel vectors), the pairwise Pearson correlation coefficient matrices are as follows: Pipe 2-5 Preset Vector Pearson Correlation Coefficient

[0067] The diagonal (autocorrelation coefficient) is always 1, and for any pipe position j, =1 if and only if = , Because each well location has a unique maximum correlation coefficient corresponding to its correct tube location, the identification is unambiguous.

[0068] Optionally, taking this N=3 channel design as an example, the Hamming distance between tube 2 (110) and tube 5 (100), and between tube 3 (101) and tube 5 (100) is 1, with the difference only in the CH2 or CH3 channel. If a well is actually filled with reagent from tube 2, but the binarization result of the CH2 channel flips from 1 to 0 due to low probe concentration, fluorescence crosstalk, or instrument noise, then the measured vector (100) will be completely consistent with the preset vector of tube 5, leading to misidentification. Quantification risk condition: Let the channel binarization flip probability be... (Single-channel false positive rate), then for a tube position pair with dH=1, the probability of false identification in a single detection is approximately ≈ For a tube pair with dH=2, the probability of false identification in a single detection is approximately 1 / 2. .therefore, =1 The reliability of the recognition design heavily depends on the accuracy of binarization. In specific detection, the following sufficient conditions must be met to guarantee a recognition accuracy of ≥99%. ,in This represents the average value including probe channels. The mean value for the probe-free channel. The standard deviation of the probe-free channel is denoted as . This condition is equivalent to requiring the fluorescence difference between the probe-enabled and probe-free channels to exceed 6 times the standard deviation of the probe-free channel, ensuring that the binarization threshold can completely separate the two types of signals.

[0069] Optionally, when channel resources permit, priority may be given to selecting Design with ≥2: For example, when N=4 channels and R≤4 tube positions, it can be from =Select 4 from 15 non-zero vectors to make ≥2, eliminating the risk of misidentification caused by single-channel flipping.

[0070] In this embodiment, maximizing the minimum Hamming distance is the optimization objective. Under the premise of meeting the detection requirements, the distinguishability between different tube position channel combinations is improved, and the distribution of each tube position code in the signal space is more discrete. This enhances the system's tolerance to actual interference factors such as fluorescence signal fluctuations, channel crosstalk, or measurement noise, and improves the stability and accuracy of automatic identification.

[0071] Based on the above embodiments, this application also provides a process for determining the target reaction tube position in the information identification method of a fluorescence detection device. Figure 4 This is a flowchart illustrating the process of determining the target reaction tube position in an information identification method for a fluorescence detection device provided in this application embodiment. Figure 4As shown, in step 103 above, based on the channel fluorescence value of each well, the preset channel combination information corresponding to the target reaction tube position is determined from the preset channel combination information of multiple reaction tube positions, including: Step 401: Calculate the fluorescence channel characteristics of each well based on the channel fluorescence value of each well.

[0072] The channel fluorescence value indicates the fluorescence statistics of each fluorescence channel in each well. The fluorescence channel feature is a feature vector composed of the features of each fluorescence channel in each well.

[0073] Optionally, the instrument background value of each fluorescence channel in each well position and the channel fluorescence value of each fluorescence channel in each well position are used to determine the net fluorescence value of each fluorescence channel in each well position.

[0074] For example, a preset net fluorescence value calculation formula is used. Based on the instrument background value of each fluorescence channel in each well position and the channel fluorescence value of each fluorescence channel in each well. The net fluorescence value of each fluorescence channel in each well was determined. For hole position index, For fluorescence channel indexing, Hole position Fluorescence channel The net fluorescence value. Hole position Fluorescence channel The channel fluorescence value, Hole position Fluorescence channel The instrument background value.

[0075] Optionally, for each well, the net fluorescence values ​​measured in each detection channel are combined into a multidimensional real vector, where each component of the vector retains its corresponding net fluorescence value.

[0076] Step 402: Calculate the similarity of channel features of each well and each reaction tube according to the fluorescence channel characteristics of each well and the preset channel combination information of each reaction tube.

[0077] Among them, channel feature similarity is the degree of matching between measured fluorescence features and preset coding features, such as Pearson correlation coefficient, cosine similarity, etc.

[0078] Optionally, based on the fluorescence channel characteristics of each well and the preset channel combination information of each reaction tube, the similarity of the channel characteristics of the well and each reaction tube is calculated using a similarity calculation formula. The similarity calculation formula can be a Pearson correlation coefficient formula, a cosine similarity formula, or a Hamming similarity formula, etc., and this application embodiment does not impose any limitations on this.

[0079] For example, using the Pearson correlation coefficient calculation formula, the calculation is performed. and Pearson correlation coefficient between As a channel feature similarity, among which The fluorescence channel characteristics of each well after treatment are shown. The channel combination information for the preset reaction tube position has a mean component of... , Pearson correlation coefficient r∈[ [1, 1], the closer the value is to 1, the higher the matching degree between the two vectors. When a channel has the same value in the preset vectors of all tube positions, the channel is excluded from the correlation calculation, and only the channels that can be effectively distinguished are calculated. Hole position Fluorescence channel feature vector Reaction tube position Preset channel combination information The Pearson correlation coefficient. Hole position Fluorescence channel characteristics of fluorescence channel i For the reaction tube position The preset characteristics of fluorescence channel i. N is the number of fluorescence channels. When the maximum correlation coefficient is reached... The second largest correlation coefficient The difference Δr= - The system outputs a "low confidence" warning and prompts for manual review instead of forcibly outputting the recognition result. θ can be 0.3.

[0080] Optionally, the cosine similarity calculation formula is used to calculate the similarity. and Cosine similarity between The cosine similarity value ranges from 0 to 1. The closer the value is to 1, the more consistent the activation patterns of the two vectors are. Hole position Fluorescence channel feature vector Reaction tube position Preset channel combination information The cosine similarity. Hole position Fluorescence channel characteristics of fluorescence channel i For the reaction tube position The preset characteristics of fluorescence channel i. N is the number of fluorescence channels.

[0081] Optionally, the Hamming distance calculation formula can be used to calculate the distance. and Hamming distance between Hamming similarity is determined based on Hamming distance. . Hole position Fluorescence channel characteristics of fluorescence channel i For the reaction tube position The preset characteristics of fluorescence channel i. N is the number of fluorescence channels.

[0082] Step 403: Based on the channel feature similarity, determine the reaction tube position corresponding to the highest similarity as the target reaction tube position corresponding to the orifice position, and determine the preset channel combination information of the target reaction tube position.

[0083] Optionally, the similarity of the channel features of the well sites and each reaction tube site is sorted from high to low, and the reaction tube site corresponding to the highest similarity is determined as the target reaction tube site corresponding to the well site, and the preset channel combination information of the target reaction tube site is determined.

[0084] In this embodiment, by constructing a similarity metric between the measured fluorescence feature vector of the well site and the preset tube channel combination, a semantic mapping from the original signal to the detection content is realized. This enables the system to perform autonomous identification based solely on the inherent optical properties of the probe, without relying on the amplification results or being affected by positive or negative results, thus improving the adaptability and reliability of the identification process.

[0085] Based on the above embodiments, this application also provides a process for determining fluorescence channel features in an information identification method for a fluorescence detection device. Figure 5 This is a flowchart illustrating the process of determining fluorescence channel features in an information identification method for a fluorescence detection device provided in an embodiment of this application, as shown below. Figure 5 As shown, in step 401 above, the fluorescence channel characteristics of each well are calculated based on the channel fluorescence value of each well, including: Step 501: Obtain the preset fluorescence threshold.

[0086] The preset fluorescence threshold refers to a fluorescence intensity boundary standard set before detection, used to distinguish whether a corresponding fluorescent probe exists in a certain detection channel. The preset fluorescence threshold can be set based on empirical values ​​of preset biological detection items, or it can be determined based on the parameters of the current fluorescence detection device; this application embodiment does not impose such limitations.

[0087] Optionally, a preset fluorescence threshold is determined based on the parameters of the fluorescence detection device or the parameters of a preset biological detection item. Alternatively, a preset fluorescence threshold is determined based on both the parameters of the fluorescence detection device and the parameters of the preset biological detection item.

[0088] Step 502: Process the channel fluorescence value of each well according to the preset fluorescence threshold to obtain the fluorescence channel characteristics of each well.

[0089] The processing can be binarization. This involves uniformly judging the channel fluorescence values ​​based on a preset fluorescence threshold. When the fluorescence value measured at a well location in a certain fluorescence channel is higher than the threshold, the fluorescence channel is considered activated, and its feature is recorded as 1. When the fluorescence value is not higher than the threshold, the fluorescence channel is considered inactive, and its feature is recorded as 0. The fluorescence channel feature refers to the result of converting the channel fluorescence value of each well location in each detection channel into a binary identifier, where the identifier indicates whether the probe is activated or not, serving as a structured expression reflecting the fluorescence channel usage status of that well location.

[0090] Optionally, the fluorescence value measured on each fluorescence channel of each well is compared with a preset fluorescence threshold. If the fluorescence value of a fluorescence channel exceeds the threshold, the fluorescence channel is determined to be activated in this well, and the feature of the fluorescence channel is recorded as 1. If the fluorescence value of a fluorescence channel does not exceed the threshold, the fluorescence channel is determined to be unactivated, and the feature of the fluorescence channel is recorded as 0. This comparison operation is performed on all wells and all fluorescence channels to obtain a vector composed of the logical values ​​of each fluorescence channel of each well, which serves as the fluorescence channel feature of that well.

[0091] For example, according to the hole position Fluorescence channel fluorescence value and fluorescence channels preset fluorescence threshold The fluorescence values ​​of each well site were binarized to obtain the fluorescence channel characteristics of each well site. Where N is the total number of fluorescence channels.

[0092] For example, according to the hole position Fluorescence channel Net fluorescence value and fluorescence channels preset fluorescence threshold The fluorescence values ​​of each well site were binarized to obtain the fluorescence channel characteristics of each well site. Where N is the total number of fluorescence channels.

[0093] For example, after full-channel binarization, the measured feature vector of the aperture position w is: .

[0094] In this embodiment, by introducing a preset fluorescence threshold and performing binarization, the fluorescence signal is converted into a stable channel activation status identifier, which suppresses interference caused by instrument noise and background fluctuations and improves the certainty and consistency of channel status discrimination.

[0095] Based on the above embodiments, this application also provides a process for determining a preset fluorescence threshold in the information recognition method of a fluorescence detection device. Figure 6 This is a schematic diagram illustrating the process of determining a preset fluorescence threshold in an information recognition method for a fluorescence detection device provided in an embodiment of this application, as shown below. Figure 6 As shown, obtaining the preset fluorescence threshold in step 501 above includes: Step 601: Obtain the instrument background value of each well and the standard deviation of the original fluorescence value of each well.

[0096] The instrument background value of a channel refers to the inherent signal baseline level of each fluorescence channel in a fluorescence detection device under conditions without samples or blank controls. It mainly originates from instrument background noise, filter leakage, and weak autofluorescence of consumables. Instrument background noise can be caused by dark current and thermal noise of photoelectric detection devices. The instrument background value of a channel is determined and stored through standardized experiments before the fluorescence detection device leaves the factory, exhibiting slight differences between well positions but relative stability between channels. The standard deviation of the original fluorescence value of a channel is the standard deviation of the original fluorescence value detected without a fluorescent probe. The standard deviation of the original fluorescence value of a channel refers to the dispersion of the original fluorescence value obtained by repeatedly measuring the same detection channel without any fluorescent probe reagents. The standard deviation of the original fluorescence value of a channel characterizes the noise fluctuation amplitude of that channel when there is no specific signal.

[0097] Optionally, the instrument background value of each well location can be obtained based on the parameters of the fluorescence detection device.

[0098] Optionally, the original fluorescence value of the channel at the detected well site without the fluorescent probe is obtained from the pre-experiment calculation, and the standard deviation is calculated using a preset formula. Calculation in multiple loops The standard deviation of the channel fluorescence of the original fluorescence value of the channel at the bottom of the well. .in, For hole position index, For fluorescence channel indexing, For the hole position Fluorescence channel In the The original fluorescence value of the channel measured during each cycle. This represents the total number of iterations. For circular index, Hole position Fluorescence channel The average original fluorescence value of the channel.

[0099] Step 602: Determine the preset fluorescence threshold of the first channel for each detection channel based on the instrument background value and the standard deviation of the original fluorescence value of each well in the same detection channel.

[0100] The preset fluorescence thresholds include: the fluorescence threshold of the first channel of each detection channel.

[0101] Optionally, based on the instrument background value of each well in the same detection channel, a preset maximum background value is used to determine the formula. Determine the maximum background value .in, For fluorescence channel indexing, For hole position index, To obtain the maximum value, For the same fluorescence channel i, the maximum background value is selected.

[0102] Optionally, based on the instrument background value of each well in the same detection channel, a preset maximum standard deviation is used to determine the formula. Determine the standard deviation of the original fluorescence value of the largest channel. .in, For fluorescence channel indexing, For hole position index, To obtain the maximum value, For the same fluorescence channel i, the standard deviation of the original fluorescence value of the largest channel is taken.

[0103] Optionally, based on the maximum background value and the standard deviation of the original fluorescence value of the largest channel The fluorescence threshold calculation formula for the first channel is used. The fluorescence threshold of the first channel is determined. The total number of channels. The fluorescence threshold of the first channel of fluorescence channel i. This represents the maximum background value of fluorescence channel i. is the standard deviation of the maximum original fluorescence value of fluorescence channel i.

[0104] Based on the above embodiments, this application also provides another process for determining a preset fluorescence threshold in the information identification method of a fluorescence detection device. In step 501 above, obtaining the preset fluorescence threshold includes: Based on the original fluorescence values ​​of each well in the same detection channel and the instrument background value of each well, the preset fluorescence threshold of the second channel for each detection channel is determined.

[0105] The preset fluorescence thresholds include: the second channel fluorescence threshold for each detection channel.

[0106] Optionally, the original net fluorescence value of each well in the same detection channel is determined based on the original fluorescence value of each well, a preset coefficient, and the instrument background value of each well. The average value of the original net fluorescence values ​​of each well in the same detection channel is used as the second channel fluorescence threshold. The average value of the original net fluorescence values ​​of each well in the same detection channel is calculated using a preset average value formula. Sure, For fluorescence channel indexing, For hole position index, The original net fluorescence value of the channel. This represents the total number of fluorescence channels. For preset coefficients, use preset coefficients To effectively distinguish between the highest net fluorescence value of the probe-free channel and the lowest net fluorescence value of the probe-enabled channel, a preset coefficient is used. The determination can be based on statistical data from multiple instruments, and this application does not impose any limitations on this.

[0107] In this embodiment, the fluorescence threshold is dynamically set by fusing the instrument’s background characteristics and the channel noise statistics. This takes into account the inherent systematic background differences between different well positions and reflects the objective distribution of the noise level of each detection channel. This makes the threshold setting physically interpretable and device adaptive, and improves the stability, consistency and cross-platform reproducibility of the binarization results.

[0108] Based on the above embodiments, this application also provides a process for determining the target reaction tube position in another information identification method for a fluorescence detection device. Figure 7 A flowchart illustrating the determination of the target reaction tube position in another information identification method for a fluorescence detection device provided in this application embodiment is shown below. Figure 7 As shown, in step 403 above, determining the reaction tube position corresponding to the highest similarity based on channel feature similarity as the target reaction tube position corresponding to the orifice position includes: Step 701: Construct a feature vector matching matrix based on the channel feature similarity of all orifices and each reaction tube.

[0109] The feature vector matching matrix is ​​a two-dimensional table data structure. Its rows represent all detected pores, and its columns represent all preset reaction tube positions. The value of each cell is the channel feature similarity between the corresponding pore and the corresponding tube position.

[0110] Optionally, the channel feature similarity between the orifice and the corresponding tube position is filled into the blank matrix according to the row and column relationship to obtain the feature vector matching matrix.

[0111] For example, fluorescence channel characteristics of R pore sites. Preset channel combination information with R reaction tube positions Calculated channel feature similarity , forming the feature vector matching matrix The similarity of channel features in the w-th row and j-th column of the feature vector matching matrix. .

[0112] Step 702: Assign values ​​to the feature vector matching matrix using a preset assignment method to obtain a new feature vector matching matrix.

[0113] The default assignment method is to find the maximum value for each row, assigning the maximum similarity value of the channel features in each row to 1, and assigning 0 to the rest. The new feature vector matching matrix includes only two features: 1 and 0.

[0114] Optionally, the feature vector matching matrix is ​​traversed row by row, with the maximum value in each row assigned a value of 1 and the rest assigned a value of 0, thereby performing the assignment process on the feature vector matching matrix and obtaining a new feature vector matching matrix.

[0115] For example, find the maximum value for each row of W, assign the maximum value a value of 1, and assign the rest a value of 0, thus forming a new feature vector matching matrix. , It is a row-based single-hot matrix. . The reaction tube position with the highest channel feature similarity. For the reaction tube location index, For the index set, in all reaction tube positions In the middle, find the correlation coefficient The biggest one , will this Recorded as . For the hole position In the matching matrix of the th In the row, only the best matching tube position is available. The corresponding column is set to 1, and all other columns in the row are set to 0.

[0116] Step 703: Based on the new feature vector matching matrix, determine the reaction tube position corresponding to the highest similarity as the target reaction tube position corresponding to the well position.

[0117] Optionally, for each well position, check which row it corresponds to in the new eigenvector matching matrix. Since each row of the eigenvector matching matrix contains only one marker with a value of one, and the rest are zero, the reaction tube position represented by the column containing the value one is the target reaction tube position.

[0118] For example, based on the new feature vector matching matrix, the reaction tube position corresponding to the highest similarity is determined as the target reaction tube position corresponding to the well position. in, For the new feature vector matching matrix, , It is an R×N matrix, which is a matrix composed of the fluorescence channel characteristics of the pore sites. as well as It is a matrix composed of preset channel combination information for the reaction tube position.

[0119] Optionally, if multiple well positions are matched with the same reaction tube position, a matching error will be indicated, and manual verification is required.

[0120] In the embodiments of this application, automatic tube position identification based on the characteristics of the fluorescence signal itself eliminates the reliance on manually preset information, physical labels or sample addition order, accurately restores the actual reagent identity loaded in each well position, and improves the fault tolerance and result traceability of multiple fluorescence detection.

[0121] Based on the above embodiments, this application also provides a process for determining channel fluorescence values ​​in an information identification method for a fluorescence detection device. Figure 8 This is a flowchart illustrating the process of determining channel fluorescence values ​​in an information identification method for a fluorescence detection device provided in this application embodiment. Figure 8 As shown, in step 102 above, fluorescence detection is performed on the well plate of the fluorescence detection device to obtain the channel fluorescence values ​​of multiple well positions on the well plate, including: Step 801: Perform multiple cycles of fluorescence detection on the well plate of the fluorescence detection device to obtain the initial channel fluorescence values ​​of multiple well positions in multiple cycles.

[0122] In each PCR cycle, for every target DNA molecule amplified, one probe is cleaved, releasing one fluorescent molecule. As the cycle number increases, the number of cleaved probes accumulates, and the fluorescence signal intensity increases exponentially, forming a typical amplification curve, which includes a baseline phase, an amplification phase, and a plateau phase. The fluorescence signal during the baseline phase is based on the background fluorescence, supplemented by the weak residual fluorescence of the probe molecules that remain in the solution throughout.

[0123] Optionally, multiple cycles of fluorescence detection are performed on the well plate of the fluorescence detection device to obtain the initial channel fluorescence values ​​of multiple well positions in multiple cycles.

[0124] Step 802: Determine the channel fluorescence values ​​of multiple wells on the plate based on the initial channel fluorescence values ​​of multiple wells in multiple cycles.

[0125] Optionally, based on the initial channel fluorescence values ​​of multiple wells over multiple cycles, a preset average value calculation formula is used to determine the average value of the initial channel fluorescence values ​​of each well over multiple cycles, which is then used as the channel fluorescence value of multiple wells on the plate. The preset average value calculation formula is as follows: ,in, Hole position Fluorescence channel The channel fluorescence value, The initial channel fluorescence value measured in channel i at well w during the c-th cycle. This represents the total number of iterations. This is a circular index.

[0126] Optionally, the initial channel fluorescence value of the last cycle among the initial channel fluorescence values ​​of multiple well sites in multiple cycles can be used as the channel fluorescence value of multiple well sites on the plate.

[0127] Optionally, the median initial channel fluorescence value among the initial channel fluorescence values ​​of multiple well sites in multiple cycles can be used as the channel fluorescence value of multiple well sites on the plate.

[0128] Optionally, the maximum initial channel fluorescence value among the initial channel fluorescence values ​​of multiple well sites in multiple cycles can be used as the channel fluorescence value of multiple well sites on the plate.

[0129] In this embodiment, the fluorescence detection process utilizes the initial fluorescence response during the initial stage before significant amplification occurs, i.e., within multiple cycles, to clearly distinguish which channels contain fluorescent probes and which do not. This is unaffected by whether the target is amplified, the amplification efficiency, or the positive or negative result, resulting in highly stable and robust channel fluorescence values. This makes the tube location identification process completely independent of the PCR amplification effect. Even if all detection targets are negative or no typical amplification curve appears, the activation status of each well location can still be accurately determined, thus ensuring that the automatic identification function is reliable and effective under various experimental conditions.

[0130] For example, this method will be explained in conjunction with specific preset biological detection items. Taking a 4-channel fluorescence detection device as an example, this invention can complete the detection of 10 pathogens and internal controls in one experiment by designing different channel combinations of 5 tube positions.

[0131] Taking a fluorescence detection device with N=4 channels for respiratory tract 10-channel detection as an example, the IC occupies CH4, and CH1-CH3 are the effective channels for detecting the target. All combinations of multi-tube 3-channel designs are shown in the table below: All 3-channel combination designs

[0132] In this specific embodiment, IC occupies CH4. Therefore, a respiratory tract 10-linked detection and 5 internal reference genes are designed. For the 4-channel fluorescence detection device, the channel combination of tube 1 is 1111, which activates channels 1, 2, 3, and 4; the channel combination of tube 2 is 1101, which activates channels 1, 2, and 4; the channel combination of tube 3 is 1011, which activates channels 1, 3, and 4; the channel combination of tube 4 is 0111, which activates channels 2, 3, and 4; and the channel combination of tube 5 is 1001, which activates channels 1 and 4. CH4 contains internal reference ICs, as shown in the table. 10-tube airway combination design

[0133] Each row in the matrix is ​​unique and has its own characteristics. Therefore, the selection of tube locations and fluorescence channels for the 10-tube multiplex fluorescent PCR reagent plus internal control IC should follow the table above.

[0134] Under the standard fluorescent PCR procedure, fluorescence detection was performed on all R wells occupied by the reaction reagents for 40 cycles, recording the entire process of fluorescence signal changes in different channels with the number of amplification cycles. Real-time fluorescent PCR was performed on well plates loaded with different items. The instrument scanned all wells according to a preset time sequence (e.g., denaturation 3s, annealing extension 15s). After annealing extension, the initial fluorescence intensity value of each well in each fluorescence channel was recorded, generating a dataset of initial fluorescence intensity values ​​for each well. The fluorescence statistics for each well in each channel were calculated; in this example, the mean fluorescence value over 40 cycles was used.

[0135] First, this application uses the correct tube sequence as an example for illustration. In this embodiment, the original fluorescence curves of 5 wells, 4 channels, and 40 cycles are shown below. Figure 9 As shown, Figure 9 This is a cycled original fluorescence curve provided for an embodiment of this application.

[0136] The average fluorescence values ​​of the 10-item respiratory tract test (5 wells, 4 channels) over 40 cycles were rounded to obtain the channel fluorescence values. The data are shown in the table below: Average fluorescence value of 40 cycles in 10-channel respiratory tract examination

[0137] The background values ​​of the borehole channels are shown in the table below: Background value of aperture channel

[0138] The fluorescence values ​​of 5 wells and 4 channels were binarized. The maximum background value of the same channel across multiple wells plus 5 times the standard deviation was used as the threshold. The binarization thresholds for each fluorescence channel are shown in the table below: Maximum background, standard deviation and binarization threshold of the channel

[0139] The fluorescence values ​​of the 4 channels at 5 wells were binarized according to the binarization threshold. The results of the fluorescence value binarization are shown in the table below: Fluorescence value binarization

[0140] For well A1, where all channels are 1, we directly search for the preset channel combination of all 1 channels in tube 1 and match it with the preset channel combination of tube 1. This means the fluorescence measurement of well A1 corresponds to the preset tube 1 FluA, FluB, and RSV+IC items. We calculate the Pearson correlation coefficient r between the eigenvectors of the fluorescence measurement channels in wells B1-E1 and the eigenvectors of the preset tubes 2-5. CH4 is a constant column and is excluded from the correlation calculation; only channels CH1-CH3 that are effectively distinguishable are calculated. The eigenvector matching matrix W and the new eigenvector matching matrix are then used. As shown in the table below: Eigenvector matching matrix

[0141] Binarizing the eigenvector matching matrix yields a new eigenvector matching matrix, as shown in the table below: New eigenvector matching matrix

[0142]

[0143] The project channel is pipe 2. (1,1,0,1), Hole B1 is the sub-detection item corresponding to preset tube 2, which is HADV, HMPV and IC item. The project channel is pipe 3 (1,0,1,1), hole C1 is the sub-detection item corresponding to preset tube 3, which is MP, HPIV1 and IC item. The project channel is pipe 4 (0,1,1,1), Hole D1 is the sub-detection item corresponding to preset tube 4, which is HPIV2, HPIV3 and IC item. The project channel is pipe 5 (1,0,0,1), hole E1 is the sub-detection item corresponding to preset tube 5, which is HPIV4 and IC item. The amplification curves of holes A1~E1 after algorithm processing are as follows. Figure 10 As shown, Figure 10This is a schematic diagram of an amplification curve provided in an embodiment of this application. The threshold line is 0.1, and the CT value is the intersection of the amplification curve and the threshold line. The CT values ​​of the 10 detection items do not intersect with the threshold line, all of which are NoCt and are all negative. The IC values ​​of A1 to E1 are all positive, with CT values ​​of 24.11, 24.62, 27.04, 28.01 and 27.78, respectively.

[0144] Based on the CT values ​​and the corresponding sub-inspection items for each hole location, the following inspection results table is provided: Project test results

[0145] Furthermore, this application uses a disordered tube arrangement as an example for illustration, specifically a 10-tube respiratory tract assay. The reagent design is as shown in the above-described 10-tube respiratory tract assay channel combination design. After adding samples to 5 tubes, they are mixed and placed together, using a 4-channel fluorescence PCR instrument as an example.

[0146] 5 wells, 4 channels, 40 cycles of original fluorescence curves, such as Figure 11 As shown, Figure 11 Another original cyclic fluorescence curve provided for an embodiment of this application. The average fluorescence values ​​for 40 cycles across 5 wells and 4 channels are shown below: Average fluorescence of 40 cycles in the orifice channel of a mixed-place tube example

[0147] The solution in this application uses the same instrument as in the above embodiments. The background value is as shown in the background value of the above-mentioned aperture channel, and the binarization threshold is as shown in the maximum background, standard deviation and binarization threshold of the above-mentioned channel.

[0148] The binarization results are shown in the table below, illustrating the binarization of fluorescence values ​​in a mixed-displacement example: Binarization of fluorescence values ​​in examples of mixed placement of tubes

[0149] In the above binarization results, the fluorescence detection channel activation feature of well E1, which is all 1s, is directly searched without calculating the correlation coefficient. It matches the preset tube position channel combination of tube 1, which is also all 1s. Therefore, the measured value of well E1 is the preset Flu A+Flu B+RSV+IC item for tube 1. The correlation coefficients of the fluorescence test channel features of wells A1~D1 with the preset tube position channel features of tubes 2~5 are calculated. CH4 is a constant column and is excluded from the correlation calculation. Only the effectively distinguishable channels CH1~CH3 are calculated. The correlation coefficient calculation results are shown in the Pearson correlation coefficient matrix W table of the fluorescence detection and preset tube position channel activation feature of the mixed tube placement example. Fluorescence measurements of mixed-place tubes and Pearson correlation coefficient matrix of preset tube channel activation characteristics W

[0150] Matching matrix as shown in the table, for example, matching matrix of mixed placement of pipes. As shown: Pipe Mixed Placement Instance Matching Matrix

[0151]

[0152] in, , The project channel is pipe 4 (0,1,1,1), hole A1 is the HPIV2+HPIV3+IC item corresponding to preset tube 4. The project channel is pipe 3 (1,0,1,1), hole B1 is the MP+HPIV1+IC item corresponding to preset tube 3. The project channel is pipe 2. (1,1,0,1), hole C1 is the HADV+HMPV+IC project corresponding to preset tube 2. The project channel is pipe 5 (1,0,0,1), hole D1 is the HPIV4+IC item corresponding to preset tube 5.

[0153] The relative fluorescence amplification curves calculated after well site matching are as follows: Figure 12 As shown, Figure 12 This is another schematic diagram of an amplification curve provided for an embodiment of this application.

[0154] Based on the CT values ​​and the corresponding sub-inspection items for each hole location, the inspection results are shown in the following table: Project Test Result Table

[0155] As can be seen, in this application, regardless of whether it is a negative amplification curve or a positive amplification curve, the method of this invention can effectively identify the preset tube position channel item based on the channel characteristics of the measured fluorescence. Although the negative amplification curve shows no amplification, the baseline fluorescence of the channel with the fluorescent probe is still higher than that of the probe-free channel, thus correctly identifying its channel activation characteristics and outputting a negative result.

[0156] Based on the same inventive concept, this application also provides an information identification device for a fluorescence detection device corresponding to the information identification method of the fluorescence detection device. Since the principle of the device in this application is similar to the information identification method of the fluorescence detection device described above in this application, the implementation of the device can refer to the implementation of the method, and the repeated parts will not be described again.

[0157] Figure 13 This is a schematic diagram of the structure of an information recognition device for a fluorescence detection device provided in an embodiment of this application, as shown below. Figure 13 As shown, the device includes: an acquisition module 1301, a detection module 1302, and a determination module 1303; wherein: The acquisition module 1301 is used to acquire the reaction tube position coding scheme of the fluorescence detection device for the preset biological detection project. The reaction tube position coding scheme includes: preset channel combination information of multiple reaction tube positions; the preset channel combination information of each reaction tube position includes: the activation detection channel corresponding to the reaction tube position and the target information of each activation detection channel; the preset channel combination information of different reaction tube positions is different. The detection module 1302 is used to perform fluorescence detection on the well plate of the fluorescence detection device during the fluorescence detection process of the preset biological detection items, and obtain the channel fluorescence values ​​of multiple well positions on the well plate; the channel fluorescence value of each well position includes: the fluorescence value of the detection channel corresponding to the well position; The determination module 1303 is used to determine the preset channel combination information of the target reaction tube position corresponding to the well position from the preset channel combination of multiple reaction tube positions based on the channel fluorescence value of each well position; and to determine the sub-detection item carried by the well position under the preset biological detection item based on the preset channel combination information.

[0158] In one possible implementation, the acquisition module 1301 is further configured to: set channels for multiple preset reaction tube positions to obtain channel combination information of multiple preset reaction tube positions, wherein the channel combination information of each preset reaction tube position includes activation indication information of at least one detection channel. Based on the information of multiple targets to be tested corresponding to the preset biological detection items, and the channel combination information of multiple preset reaction tube positions, the reaction tube position coding scheme is determined.

[0159] In one possible implementation, the acquisition module 1301 is specifically used to: set channels for multiple preset reaction tube positions according to preset non-zero constraint conditions and uniqueness constraint conditions, and obtain channel combination information of multiple preset reaction tube positions; the preset non-zero constraint conditions are used to indicate that each preset reaction tube position activates at least one detection channel, and the uniqueness constraint conditions are used to indicate that the channel combination information of different preset reaction tube positions is different.

[0160] In one possible implementation, the target information to be tested includes: the total number of targets and the type of each target; the acquisition module 1301 is specifically used to: calculate the Hamming distance between every two channel combinations in the channel combination information of multiple preset reaction tube positions, and determine the minimum Hamming distance; Based on the total number of targets, the required number of preset reaction tube positions, and the type of each target, the set of channel combinations with the largest minimum Hamming distance is selected from the channel combination information of multiple preset reaction tube positions as the reaction tube position coding scheme, using the constraint of the total number of preset targets. Among them, the preset target total number constraint condition is used to indicate that the number of activation detection channels of all reaction tube positions in the reaction tube position coding scheme is equal to the total number of targets.

[0161] In one possible implementation, the determining module 1303 is specifically used to: calculate the fluorescence channel characteristics of each well based on the channel fluorescence value of each well. Based on the fluorescence channel characteristics of each well and the preset channel combination information of each reaction tube, the similarity of channel characteristics of the well and each reaction tube is calculated respectively. Based on the channel feature similarity, the reaction tube position corresponding to the highest similarity is determined as the target reaction tube position corresponding to the orifice position, and the preset channel combination information of the target reaction tube position is determined.

[0162] In one possible implementation, the determining module 1303 is specifically used to: obtain a preset fluorescence threshold; Based on a preset fluorescence threshold, the channel fluorescence values ​​of each well are processed to obtain the fluorescence channel characteristics of each well.

[0163] In one possible implementation, the determining module 1303 is specifically used to: obtain the instrument background value of the channel at each well and the standard deviation of the original fluorescence value of the channel at each well; wherein, the standard deviation of the original fluorescence value of the channel is the standard deviation of the channel fluorescence corresponding to the original fluorescence value of the channel when no fluorescent probe is placed; Based on the instrument background value and the standard deviation of the original fluorescence value of each well in the same detection channel, a preset fluorescence threshold for the first channel is determined for each detection channel. The preset fluorescence threshold includes: the first channel fluorescence threshold for each detection channel; or... Based on the original fluorescence value of each well in the same detection channel, the preset coefficient, and the instrument background value of each well, a second channel preset fluorescence threshold is determined for each detection channel. The preset fluorescence threshold includes the second channel fluorescence threshold for each detection channel.

[0164] In one possible implementation, the determining module 1303 is specifically used to: construct a feature vector matching matrix based on the channel feature similarity of all orifice locations and each reaction tube location; The matching matrix is ​​assigned values ​​using a preset assignment method to obtain a new feature vector matching matrix; Based on the new feature vector matching matrix, the reaction tube position corresponding to the highest similarity is determined as the target reaction tube position corresponding to the well position.

[0165] In one possible implementation, the detection module 1302 is specifically used to: perform multiple cycles of fluorescence detection on the well plate of the fluorescence detection device to obtain the initial channel fluorescence values ​​of multiple well positions in multiple cycles; The channel fluorescence values ​​of multiple wells on the plate are determined based on the initial channel fluorescence values ​​of multiple wells in multiple cycles.

[0166] The processing flow of each module in the device and the interaction flow between each module can be referred to the relevant descriptions in the above method embodiments, and will not be detailed here.

[0167] This application also provides a hardware system. Figure 14 This application provides a schematic diagram of the structure of a hardware system, as shown in the embodiment. Figure 14 As shown, the system includes a processor 1401 and a memory 1402, and optionally, a bus 1403. The memory 1402 stores machine-readable instructions executable by the processor 1401. When the hardware system is running, the processor 1401 and the memory 1402 communicate via the bus 1403. When the machine-readable instructions are executed by the processor 1401, the steps of the information recognition method for the fluorescence detection device described above are performed. The hardware system is a device connected to the fluorescence detection device for processing the detection data of the fluorescence detection device.

[0168] This application also provides a computer-readable storage medium storing a computer program, which, when executed by a processor, performs the steps of the information recognition method of the fluorescence detection device described above.

[0169] Those skilled in the art will clearly understand that, for the sake of convenience and brevity, the specific working processes of the systems and devices described above can be referred to the corresponding processes in the method embodiments, and will not be repeated here. In the several embodiments provided in this application, it should be understood that the disclosed systems, devices, and methods can be implemented in other ways. The device embodiments described above are merely illustrative. For example, the division of modules is only a logical functional division, and in actual implementation, there may be other division methods. Furthermore, multiple modules or components can be combined or integrated into another system, or some features can be ignored or not executed. Another point is that the displayed or discussed mutual coupling or direct coupling or communication connection can be through some communication interfaces; the indirect coupling or communication connection of devices or modules can be electrical, mechanical, or other forms.

[0170] Furthermore, the functional units in the various embodiments of this application can be integrated into one processing unit, or each unit can exist physically separately, or two or more units can be integrated into one unit. If the functions are implemented as software functional units and sold or used as independent products, they can be stored in a computer-readable storage medium. Based on this understanding, the technical solution of this invention, or the part that contributes to the prior art, or a part of the technical solution, can be embodied in the form of a software product. This computer software product is stored in a storage medium and includes several instructions to cause a hardware system (which may be a personal computer, server, or network device, etc.) to execute all or part of the steps of the methods described in the various embodiments of this invention. The aforementioned storage medium includes: USB flash drives, portable hard drives, read-only memory (ROM), random access memory (RAM), magnetic disks, optical disks, and other media capable of storing program code.

[0171] The above are merely specific embodiments of this application, but the scope of protection of this application is not limited thereto. Any changes or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in this application should be included within the scope of protection of this application.

Claims

1. A method for information identification of a fluorescence detection device, characterized in that, The method includes: The reaction tube position coding scheme of the fluorescence detection device for the preset biological detection project is obtained. The reaction tube position coding scheme includes: preset channel combination information of multiple reaction tube positions; the preset channel combination information of each reaction tube position includes: the activation detection channel corresponding to the reaction tube position and the target information of each activation detection channel; the preset channel combination information of different reaction tube positions is different. During the fluorescence detection process of the preset biological detection item, fluorescence detection is performed on the well plate of the fluorescence detection device to obtain the channel fluorescence values ​​of multiple well positions on the well plate; the channel fluorescence value of each well position includes: the fluorescence value of the detection channel corresponding to the well position; Based on the channel fluorescence value of each well, the preset channel combination information of the target reaction tube corresponding to the well is determined from the preset channel combinations of the multiple reaction tube positions; Based on the preset channel combination information, determine the sub-detection items carried by the pore position under the preset biological detection items.

2. The method according to claim 1, characterized in that, Before acquiring the reaction tube location coding scheme for the preset biological detection item using the fluorescence detection device, the method further includes: Channel settings are performed on multiple preset reaction tube positions to obtain channel combination information for multiple preset reaction tube positions. The channel combination information for each preset reaction tube position includes activation indication information for at least one detection channel. Based on the information of multiple targets to be tested corresponding to the preset biological detection items, and the channel combination information of multiple preset reaction tube positions, the reaction tube position coding scheme is determined.

3. The method according to claim 2, characterized in that, The process of setting channels for multiple preset reaction tube positions to obtain channel combination information for the multiple preset reaction tube positions includes: Based on preset non-zero constraint conditions and uniqueness constraint conditions, channel settings are performed on multiple preset reaction tube positions to obtain channel combination information of multiple preset reaction tube positions; the preset non-zero constraint conditions are used to indicate that each preset reaction tube position activates at least one detection channel, and the uniqueness constraint conditions are used to indicate that the channel combination information of different preset reaction tube positions is different.

4. The method according to claim 2, characterized in that, The target information includes: the total number of targets and the type of each target; determining the reaction tube location coding scheme based on the target information corresponding to the preset biological detection item and the channel combination information of multiple preset reaction tube locations includes: Calculate the Hamming distance between every two channel combinations in the channel combination information of the multiple preset reaction tube positions, and determine the minimum Hamming distance; Based on the total number of targets, the required number of preset reaction tube positions, and the type of each target, the set of channel combinations with the largest minimum Hamming distance is selected from the channel combination information of multiple preset reaction tube positions as the reaction tube position coding scheme, using the preset target total number constraint condition. The preset target total number constraint condition is used to indicate that the number of activation detection channels for all reaction positions in the reaction position coding scheme is equal to the total number of targets.

5. The method according to claim 1, characterized in that, The step of determining the preset channel combination information of the target reaction tube position corresponding to the well position from the preset channel combination information of multiple reaction tube positions based on the channel fluorescence value of each well position includes: Based on the channel fluorescence value of each pore, calculate the fluorescence channel characteristics of each pore. Based on the fluorescence channel characteristics of each well site and the preset channel combination information of each reaction tube site, the channel feature similarity between the well site and each reaction tube site is calculated respectively. Based on the channel feature similarity, the reaction tube position corresponding to the highest similarity is determined as the target reaction tube position corresponding to the orifice position, and the preset channel combination information of the target reaction tube position is determined.

6. The method according to claim 5, characterized in that, The step of calculating the fluorescence channel characteristics of each well site based on the channel fluorescence value of each well site includes: Obtain the preset fluorescence threshold; Based on the preset fluorescence threshold, the channel fluorescence value of each pore is processed to obtain the fluorescence channel characteristics of each pore.

7. The method according to claim 5, characterized in that, The process of obtaining the preset fluorescence threshold includes: Obtain the instrument background value of the channel at each well location and the standard deviation of the original fluorescence value of the channel at each well location; wherein, the standard deviation of the original fluorescence value of the channel is the standard deviation of the channel fluorescence corresponding to the original fluorescence value of the channel when no fluorescent probe is placed; Based on the instrument background value of each aperture in the same detection channel and the standard deviation of the original fluorescence value of the channel, a first channel preset fluorescence threshold is determined for each detection channel. The preset fluorescence threshold includes: the first channel fluorescence threshold for each detection channel; or... Based on the original fluorescence value of each well in the same detection channel, the preset coefficient, and the instrument background value of each well, a second channel preset fluorescence threshold is determined for each detection channel. The preset fluorescence threshold includes the second channel fluorescence threshold for each detection channel.

8. The method according to claim 5, characterized in that, The step of determining the reaction tube position corresponding to the highest similarity based on the channel feature similarity as the target reaction tube position corresponding to the aperture position includes: Based on the channel feature similarity of all the said orifice locations and each of the said reaction tube locations, a feature vector matching matrix is ​​constructed; The feature vector matching matrix is ​​assigned values ​​using a preset assignment method to obtain a new feature vector matching matrix. Based on the new feature vector matching matrix, the reaction tube position corresponding to the highest similarity is determined as the target reaction tube position corresponding to the well position.

9. The method according to claim 1, characterized in that, The step of performing fluorescence detection on the well plate of the fluorescence detection device to obtain the channel fluorescence values ​​of multiple well positions on the well plate includes: The fluorescence detection device is subjected to multiple cycles of fluorescence detection on the well plate to obtain the initial channel fluorescence values ​​of multiple well positions in multiple cycles; The channel fluorescence values ​​of multiple well sites on the plate are determined based on the initial channel fluorescence values ​​of multiple well sites in multiple cycles.

10. A hardware system, characterized in that, include: The processor and memory, the memory storing machine-readable instructions executable by the processor, which, when the hardware system is running, are executed by the processor to perform the steps of the information identification method of the fluorescence detection device as described in any one of claims 1 to 9.

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