Rapid detection method for microorganism ATP (adenosine triphosphate) based on feeding milk product

Abstract

The invention relates to the technical field of dairy product quality detection, in particular to a method for rapidly detecting microorganism ATP (adenosine triphosphate) based on feeding milk products, which comprises the following steps: collecting volume readings based on feeding milk samples, screening and distributing numbers, reading lactose components, marking and classifying, and matching lactase addition amount with reaction set values. Introducing a luminous enzyme solution, setting a heating control point, and recording fluorescence change to generate a reaction intensity classification result. According to the method, through continuous operation links such as dairy product detection, distribution, recognition, action, setting and response, accurate control of sample volume, lactose components, reaction temperature and enzyme reaction time is realized, and dynamic monitoring and accurate classification of lactose hydrolysis and ATP reaction processes are ensured in combination with fluorescence signal intensity change section division and reaction information recording; the judgment efficiency and classification accuracy of lactose existence and metabolic reaction activity are effectively improved, and the precise detection requirements of special dairy products for infants on quality stability and nutritional ingredient controllability are met.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of dairy product quality detection, in particular to a rapid detection method for microorganisms in infant milk products based on ATP. BACKGROUND

[0002] The technical field of dairy product quality detection involves comprehensive monitoring and analysis of quality and safety indicators during the production, storage, transportation and sale of dairy products. Its core tasks include microorganism content detection, chemical composition analysis, adulteration identification and physicochemical indicator detection. This technical field focuses on real-time or laboratory detection of dairy product quality through physical, chemical and biological means to ensure its safety and nutritional value. Among the numerous detection indicators, microorganism detection is an important parameter for determining whether dairy products meet food safety standards, especially in infant milk products such as infant formula milk. This field widely uses bacterial culture method, molecular biology analysis method, immunology analysis method and bioluminescence method to evaluate microorganism contamination. Bioluminescence method is gradually gaining attention due to its simple operation and fast detection speed. Traditional microorganism detection in infant milk products involves counting microbial colonies on culture medium after sample culture to determine the microorganism content in dairy products. It analyzes whether there are excessive bacteria and other microorganism indicators in dairy products. It uses microbial culture techniques such as plate spreading, pouring and filter membrane method for testing. It observes microbial growth under specific culture time and temperature conditions and uses colony forming units as the basis for judgment.

[0003] Existing technologies rely on microbial colony culture to detect dairy products in actual operation. They are highly dependent on culture time and temperature due to the growth conditions of microbial colonies, making it difficult to achieve efficient response and accurate classification within a short time. They lag in identifying lactose components or fast-acting substances, especially in infant milk products, where they do not respond promptly to trace changes in indicators. This poses a risk of insufficient lactose identification or enzyme hydrolysis. When components fluctuate during transportation, traditional colony observation method cannot dynamically track lactose activity and its change process, leading to delayed detection results, ambiguous judgment criteria and other problems. SUMMARY

[0004] In order to solve the technical problems that the existing technology depends on the culture of microbial colonies for detecting dairy products in actual operation, is highly dependent on the culture time and temperature of the colony growth conditions, is difficult to achieve efficient response and accurate classification in a short time, lags behind in the identification of lactose components or fast reactants, especially in the reaction of trace changes in infant milk, and causes the risk of insufficient identification of lactose or insufficient enzymolysis, and the traditional colony observation method is difficult to dynamically track the lactose activity and its change process when the components fluctuate in transportation, resulting in lagging detection results and fuzzy determination standard, the embodiment of the present application provides a rapid detection method for microorganisms in infant milk product based on ATP. The technical solution is as follows: In one aspect, a rapid detection method for microorganisms in infant milk product based on ATP is provided, which comprises: S1: Based on the liquid sample of the infant milk product, introducing a separate sample unit of a multi-channel liquid distribution device, sequentially collecting the sample volume reading in the sample unit, classifying and judging with the distribution control threshold area, and generating a sample distribution channel number set; S2: Through the sample distribution channel number set, the corresponding channel sample is injected into the temperature control adjustment unit, the initial temperature reading and the sample lactose component reading are read, the lactose reading is compared with the lactose judgment reference range, and a lactose judgment channel label table is generated; S3: According to the lactose judgment channel label table, the channel number including lactose is extracted in sequence, the lactose reading is called and the corresponding lactase adding volume is matched, the enzyme adding sequence is configured and then the sample is introduced, according to the sample reaction temperature, through a preset temperature-time function relationship, the lactase action time setting value is determined, and a lactose hydrolysis execution list is generated; S4: Calling the lactose hydrolysis execution list, introducing the luminescent enzyme solution into the corresponding channel sample according to the time node after hydrolysis is completed, setting the temperature control point and stable time section of the temperature control adjustment device, marking the temperature adjustment completion state under each channel, and generating a temperature control matching completion number set.

[0005] As a further scheme of the present application, the sample distribution channel number set comprises a channel number set, a distribution volume control number, and a sample unit number, the lactose judgment channel label table comprises a lactose signal classification label, a channel lactose detection state, and a channel number annotation result, the lactose hydrolysis execution list comprises lactase adding volume information, lactase action time setting value, and sample reaction temperature setting parameter, and the temperature control matching completion number set comprises a channel temperature control completion state identifier, a temperature control point number, and a stable time section identifier.

[0006] As a further scheme of the present application, the acquisition step of the sample distribution channel number set is specifically: S101: Based on the feeding milk product liquid sample, introduce the independent sample unit of the multi-channel liquid distribution device, sequentially collect the sample volume reading in the sample unit, call the volume sensing component connected to the sample unit in the multi-channel liquid distribution device, obtain the real-time volume sampling data of the channel, and generate a channel volume data set; S102: According to the channel volume data set, call the volume distribution control threshold set in the sample distribution control, compare the volume value in the sample unit with the upper and lower limit values of the volume distribution control threshold set by item, screen the sample unit number whose volume value is in the interval range, and recombine the number and the corresponding volume value in the screening result to obtain a screening sample channel reference value list; S103: Using the screening sample channel reference value list, assign an independent volume control number to the sample unit number, pair the number with the original number in the sample unit one by one, and bind the volume control number and the original sample number to the sample distribution channel together to generate a sample distribution channel number set.

[0007] As a further scheme of the present application, the acquisition step of the lactose determination channel label table is specifically: S201: Based on the sample distribution channel number set, sequentially inject the channel sample corresponding to the sample distribution channel number set into the temperature control adjustment unit, call the sample injection interface and the internal temperature control groove of the temperature control unit, complete the sample injection by number control sequence, read the initial temperature value when the channel sample is injected, uniformly record the temperature parameter data, and generate a sample channel temperature reading set; S202: According to the channel number information in the sample channel temperature reading set, collect the lactose component reading in the channel sample, uniformly arrange the lactose reading to the corresponding channel number, establish a one-to-one correspondence structure between the channel number and the lactose reading, and obtain a channel lactose reading value set; S203: According to the lactose reading in the channel lactose reading value set, call the upper and lower limit values of the lactose judgment reference range, compare the lactose reading corresponding to the channel with the reference range in the interval, judge whether there is a detectable lactose signal in the reference interval, and mark the judgment result according to the channel number to generate a channel lactose judgment mark set; S204: Based on the mark state in the channel lactose judgment mark set, call the number information in the sample distribution channel number set, bind and combine the channel number and the corresponding lactose determination state, classify according to the determination result, mark as two types of labels: detectable or undetectable, summarize the label information of the channel, and generate a lactose determination channel label table.

[0008] As a further scheme of the present application, the acquisition step of the lactose hydrolysis execution list is specifically: S301: Based on the lactose judgment channel label table, the channel number marked as a detectable lactose signal is extracted in sequence, the lactose reading under the corresponding channel is called, and the lactose reading is matched with the set lactase addition volume to generate a channel lactase volume value table; S302: According to the channel lactase volume value table, the addition order of lactase is arranged in sequence according to the channel number, the enzyme addition instruction is sequentially introduced into the corresponding channel sample, the lactase action time setting value under the enzyme addition state is collected, and the setting value under the differential channel is numbered and marked to obtain a channel lactase action time table; S303: Using the action time value in the channel lactase action time table, combined with the sample reaction temperature value read in the real-time temperature control unit, the channel temperature adjustment weight value is calculated, the corresponding combination is performed according to the channel number, the lactase action time and the corresponding temperature parameter are combined, and a lactose hydrolysis execution list is generated.

[0009] As a further scheme of the present application, the channel temperature adjustment weight value is a numerical index for measuring the influence degree of temperature fluctuation in the reaction channel on the enzyme reaction efficiency.

[0010] As a further scheme of the present application, the acquisition step of the temperature control matching completion number set is specifically: S401: Call the channel lactase action time and reaction temperature parameter in the lactose hydrolysis execution list, identify the time node after hydrolysis is completed, introduce the luminescent enzyme solution into the corresponding channel sample in sequence according to the channel number, record the introduction time and channel number, and integrate the channel introduction state to generate a channel luminescent enzyme introduction record set; S402: Based on the time node information in the channel luminescent enzyme introduction record set, set the temperature control adjustment equipment under the corresponding channel to the temperature control point and the stable time section, judge whether the set temperature is reached and maintained for the set stable time length channel by channel, mark the channels meeting the conditions as temperature adjustment completion state, and generate a temperature control matching completion number set.

[0011] As a further scheme of the present application, the channel lactase action time is the duration required for lactase to react with lactose in the detection channel; The reaction temperature parameter is a temperature control value corresponding to the lactase action time.

[0012] As a further scheme of the present application, the method further comprises a S5 step: S5: Based on the temperature control matching completion number set, activate the ATP reaction of the luminescent enzyme system under the multi-channel, record the change of fluorescence signal intensity in each channel, divide the fluorescence response intensity curve according to the change of time, record the reaction information of the channel response section, and generate a sample ATP reaction intensity classification result; The sample ATP reaction intensity classification result includes fluorescence signal intensity time change data, response section reaction information set, and channel response intensity level label.

[0013] As a further scheme of the present application, the acquisition step of the sample ATP reaction intensity classification result is specifically: S501: Adopting the channel numbers listed in the temperature control matching complete number set, activating the luminescent enzyme system ATP reaction under the multi-channel, recording the fluorescence signal intensity data under the channel according to the reaction condition of the triggered luminescent enzyme and the substrate in the sample, and generating a fluorescence intensity change data set; S502: According to the fluorescence intensity change data set, dividing the channel fluorescence signal according to the time axis, integrating the signal intensity change sequence with time into a continuous response curve, calling the channel number information to classify and arrange the curve, and extracting the start and end time and corresponding intensity value of the change section in the response curve to obtain a channel fluorescence response section value set; S503: Based on the channel fluorescence response section value set, using the maximum-minimum normalization method to normalize the response intensity, comparing the normalized response amplitude with the preset intensity threshold to classify the channel into high, medium and low intensity categories, and generating a sample ATP reaction intensity classification result.

[0014] The technical scheme provided by the embodiment of the present application has at least the following beneficial effects: By detecting dairy products, distributing, identifying, acting, setting and responding to continuous operation links, the sample volume, lactose composition, reaction temperature and enzyme reaction time are accurately controlled, and the fluorescence signal intensity change section is divided and the reaction information is recorded, so that the dynamic monitoring and accurate classification of lactose hydrolysis and ATP reaction process are ensured, the determination efficiency and classification accuracy of lactose existence and metabolic reaction activity are effectively improved, the synergistic closed loop of sample screening, enzyme addition and temperature control adjustment is realized, the false sampling and detection distortion are avoided, the identification sensitivity of suspicious lactose composition in dairy products and the quantitative tracking ability of reaction process are significantly enhanced, and the precise detection needs of quality stability and controllability of nutritional ingredients of infant special milk are met. BRIEF DESCRIPTION OF DRAWINGS

[0015] Figure 1 It is a workflow diagram of the present application; Figure 2 It is an S1 detailed flowchart of the present application; Figure 3 It is an S2 detailed flowchart of the present application; Figure 4 It is an S3 detailed flowchart of the present application; Figure 5 It is an S4 detailed flowchart of the present application; Figure 6 This is a detailed flowchart of S5 of the present invention. Detailed Implementation

[0016] The technical solution of the present invention will now be described with reference to the accompanying drawings.

[0017] In embodiments of the present invention, words such as "exemplarily," "for example," etc., are used to indicate that something is an example, illustration, or description. Any embodiment or design described as "exemplary" in the present invention should not be construed as being more preferred or advantageous than other embodiments or designs. Specifically, the use of the word "exemplary" is intended to present the concept in a concrete manner. Furthermore, in embodiments of the present invention, the meaning expressed by "and / or" can be both, or either one.

[0018] To make the technical problems, technical solutions and advantages of the present invention clearer, a detailed description will be given below in conjunction with the accompanying drawings and specific embodiments.

[0019] Please see Figure 1 This invention provides a rapid detection method for microbial ATP in breast milk products. The processing flow of this method may include the following steps: S1: Based on the liquid samples of breast milk products, an independent sample unit of a multi-channel liquid dispensing device is introduced. The sample volume readings in the sample unit are collected sequentially, classified and judged with the dispensing control threshold zone, the sample unit number in the dispensing interval is selected, the corresponding volume value is recorded and the volume control number is assigned, and a sample dispensing channel number set is generated. S2: By using the sample allocation channel number set, the corresponding channel samples are injected into the temperature control unit respectively. The initial temperature reading and sample lactose content reading are read. The lactose reading is compared with the lactose judgment reference range to determine whether there is a detectable lactose signal in the channel. The lactose judgment channel label table is generated by combining the channel number label classification. S3: Based on the lactose determination channel label table, extract the channel numbers including lactose in sequence, call the lactose reading and match the corresponding lactase addition volume, configure the enzyme addition order and import the sample, determine the lactase action time setting value according to the sample reaction temperature through the preset temperature-time function relationship, and generate the lactose hydrolysis execution list; S4: Call the lactose hydrolysis execution list, import the luminescent enzyme solution into the corresponding channel sample according to the time node after hydrolysis is completed, set the temperature control point and stabilization time segment of the temperature control device, mark the temperature regulation completion status of each channel, and generate a temperature control matching completion number set. S5: based on the temperature control matching complete number set, activate the multi-channel ATP reaction of the light enzyme system, record the fluorescence signal intensity change in each channel, divide the fluorescence response intensity curve according to the time change, record the reaction information of the channel response section, and generate the sample ATP reaction intensity classification result; The sample distribution channel number set includes a channel number set, an allocation volume control number, and a sample unit number. The lactose determination channel label table includes a lactose signal classification label, a channel lactose detection state, and a channel number annotation result. The lactose hydrolysis execution list includes lactase addition volume information, lactase action time setting value, and sample reaction temperature setting parameter. The temperature control matching complete number set includes channel temperature control completion state identification, temperature control point number, and stable time section identification. The sample ATP reaction intensity classification result includes fluorescence signal intensity time change data, response section reaction information set, and channel response intensity level label.

[0020] Please refer to Figure 2 The acquisition steps of the sample distribution channel number set are as follows: S101: based on the infant milk product liquid sample, introduce the independent sample unit of the multi-channel liquid distribution device, sequentially collect the sample volume reading in the sample unit, call the volume sensing component connected to the sample unit in the multi-channel liquid distribution device, acquire the real-time volume sampling data of the channel, and generate the channel volume data set; The independent sample unit of the multi-channel liquid distribution device needs to be introduced by manual or automatic infusion. Each milk sample is injected into the independent sample unit set in the multi-channel device. Each sample unit is provided with a unique number for tracking and tracing. The device has 12 channels, which can be set as C001 to C012. After completion, start the volume acquisition device equipped in the device, call the volume sensing component connected to each sample unit through sequential polling to obtain the current sample liquid volume information. The sensor uses differential pressure, float or electronic measurement technology for real-time volume detection. Each sampling needs to be combined with sampling timestamp record to ensure the timeliness and traceability of the data. For example, each sampling is set at 5 second intervals, sequentially from C001 to C012. After obtaining the current sample volume value of each channel, the device will arrange and summarize the volume values of each channel to form a data set corresponding to the channel and volume value. Set C001 to 1.4 mL, C002 to 1.6 mL, etc. Record and store in the database to generate the channel volume data set.

[0021] S102: According to the channel volume data set, the volume allocation control threshold set in the sample allocation control is called, the volume value in the sample unit is compared with the upper and lower limit values of the volume allocation control threshold, the sample unit number in the interval range is screened, and the number and the corresponding volume value in the screening result are recombined to obtain a screened sample channel control value list; The volume allocation control threshold set in the device is called for data screening, the upper and lower limit values of the threshold are set according to the requirements of the sample analysis and the accuracy of the device, the minimum volume allocation control value is set to 1.4 mL, and the maximum control value is set to 1.8 mL. Each item in the sample volume data set needs to be compared with the upper and lower limits, and the execution logic is checked item by item in the order of channel number. The device reads the volume value corresponding to each channel number and compares it with the above threshold. If the volume of channel C003 is 1.5 mL, it is retained because it is in the set interval. If the volume of channel C004 is 1.2 mL, it is not retained because it does not meet the condition. After screening, the device recombines the retained channel numbers and corresponding volume values. Not only the sample unit numbers that pass the screening are recorded, but also the corresponding volume values are retained for subsequent number allocation and control device calling. The whole screening process strictly executes the control logic to ensure that only sample units that meet the control standard are retained, and a screened sample channel control value list is obtained.

[0022] S103: Using the screened sample channel control value list, an independent volume control number is allocated to the sample unit number, which is paired with the original number one by one, and the volume control number and the original sample number are bound together in the sample allocation channel to generate a sample allocation channel number set; The device allocates an independent volume control number to each sample unit number, which is generated in the order of the screening result. The first qualified sample allocation is set to V001, the second is set to V002, and so on. This number is mainly used for subsequent liquid control logic execution and tracking. After the device completes the number generation, it pairs and binds the volume control number with the original sample number. Channel C005 corresponds to sample number Sample_05, and volume control number V003 is generated. The combination relationship Sample05-C005-V003 is formed by binding, which is written into the sample allocation device database for subsequent sample control execution in the actual allocation process, including allocation path, liquid quantity control, sample identification, data recording. Each number can be used as a unique identification mark. The device calls the number set information to perform sample allocation task allocation operation, ensures that each operation step in the subsequent processing process can be clearly positioned, and forms a complete data loop of the device to generate a sample allocation channel number set.

[0023] Referring to Figure 3 The acquisition step of the lactose determination channel label table is specifically as follows: S201: Based on the sample distribution channel number set, the channel samples corresponding to the sample distribution channel number set are sequentially injected into the temperature control adjustment unit, the sample injection interface and the internal temperature control groove of the temperature control unit are called, the sample injection is sequentially completed through number control, the initial temperature value when the channel sample is injected is read, the temperature parameter data is uniformly recorded, and a sample channel temperature reading set is generated; The sample unit corresponding to each channel is sequentially introduced into the temperature control groove in the temperature control adjustment unit, the injection operation is sequentially executed by reading the number sequence, the channel numbers such as T01, T02, etc. control the injection sequence according to the arrangement sequence, the temperature control groove has multiple independent interfaces, each interface is paired with a sample channel, when a channel sample is injected, the initial temperature value of the current sample entering the groove is recorded in real time by the built-in sensing module of the temperature control adjustment device, which is used as a reference parameter for subsequent temperature adjustment and is uniformly recorded in the data record table. The reading method is to read the temperature once for every injected sample, and the reading frequency is set to every 2 seconds to prevent temperature fluctuations during injection from affecting data consistency. The temperature reading after injecting the first channel sample T01 is 24.6°C, and the temperature reading after injecting the second channel sample T02 is 25.1°C. Each temperature record corresponds to a sample number and is written into the sample temperature reading table. The temperature control groove maintains a set environmental temperature range of 25±0.5°C. If the temperature exceeds the upper and lower limits during injection, the current data is marked as an abnormal record. The temperature recording data accuracy is controlled to be 0.1°C, and a sample channel temperature reading set is generated.

[0024] S202: According to the channel number information in the sample channel temperature reading set, the lactose component reading corresponding to the channel sample is collected, the lactose reading is uniformly arranged under the corresponding channel number, a one-to-one correspondence structure of channel number and lactose reading is established, and a channel lactose reading value set is obtained; The lactose component collection operation is performed on the sample unit injected into each channel. The lactose reading is obtained by molecular spectroscopy or biochemical reagent reaction. The detection function is triggered one by one according to the channel number through a specific detection module. After each detection is completed, the obtained lactose reading is paired and stored with the channel number. After the channel completes the detection, the lactose data is arranged and stored in classification according to the number structure, forming a complete channel lactose reading value set. The lactose content of T01 sample is set to 3.8g / 100mL, the lactose content of T02 is set to 4.1g / 100mL, and the lactose content of T03 is set to 3.5g / 100mL. The lactose reading accuracy is set to 0.01g / 100mL. In the arrangement process, the channel number is uniformly used as the index field, and the lactose data field is hung under each field to obtain the channel lactose reading value set.

[0025] S203: According to the lactose reading in the channel lactose reading value set, the upper and lower limit values of the lactose judgment reference range are called, the lactose reading corresponding to the channel is compared with the reference range, it is judged whether the channel exists in the reference interval The detectable lactose signal is located, and the judgment result is marked according to the channel number to generate a channel lactose judgment mark set; The reference range comparison is performed on the lactose reading under each number, the artificial setting of lactose reference interval is used as the comparison standard, the lactose reference range is set to 3.6 to 4.2g / 100mL, the channel-by-channel comparison operation is performed, the lactose reading of each channel is judged with the interval, if the lactose value falls within the interval, the channel is marked as "effective", otherwise it is marked as "invalid", the one-way comparison logic is used to execute the judgment operation in sequence according to the channel, each comparison result immediately generates a mark item and is attached below the channel number, T01 is set to 3.8g / 100mL, marked as "effective", T03 is 3.5g / 100mL, marked as "invalid", after all numbers are judged, the mark content is written into the lactose judgment mark set.

[0026] S204: Based on the mark state in the channel lactose judgment mark set, the number information in the sample distribution channel number set is called, the channel number and the corresponding lactose judgment state are bound and combined, and are classified according to the judgment result, and are marked as two types of labels, namely detectable and undetectable, the label information of the channel is summarized, and a lactose judgment channel label table is generated; Each sample distribution channel number is bound with the corresponding judgment mark, the data pairing process is performed by reading the judgment mark state field and the channel number field, after pairing, the judgment result is classified, all samples marked as "effective" are classified into "detectable" category, and all samples marked as "invalid" are classified into "undetectable" category, forming two sample sets, the corresponding labels are recorded as "detectable" and "undetectable", respectively, the two sets are reordered and summarized, taking the channel number as the primary key field, and the corresponding record label state field, the data in the table is arranged in the order of sample detection completion, which can support classification reading according to number or label type, the detectable category includes T01, T02, T04, etc., the undetectable category includes T03, T05, T06, etc., the initial lactose reading, judgment result and label in each record need to be retained for subsequent quality analysis, data verification and sample distribution logic optimization, and a lactose judgment channel label table is generated.

[0027] Please refer to Figure 4 , the acquisition steps of the lactose hydrolysis execution list are as follows: S301: Based on the lactose judgment channel label table, the channel numbers marked as detectable lactose signal are extracted in sequence, the lactose reading under the corresponding channel is called, and the lactose reading is matched with the set lactase addition volume to generate a channel lactase volume value table; Extract the channel number labeled as "detectable" one by one, set T01, T02, T04, etc., call the corresponding lactose reading data with the channel number as the index item, set T01 lactose reading as 3.8 g / 100 mL, T02 as 4.0 g / 100 mL, according to the lactose content and lactase ratio setting rule, match the lactase volume value to be added, and convert the addition ratio according to the preset fixed reference value, set 0.5 mL of lactase liquid per 1 g of lactose, then T01 should add 1.9 mL of enzyme volume, T02 should add 2.0 mL, the addition amount of the channel needs to be matched under the unified standard, and the enzyme volume value is retained to two decimal places, after the processing is completed, the lactose reading and the enzyme addition volume form the corresponding relationship of the channel, and the record format is set as: channel number-lactose reading-lactase addition volume three items in a group, set T01-3.8-1.9, and the channels are sorted and arranged according to the channel number to form a channel lactase volume value table.

[0028] S302: According to the channel lactase volume value table, arrange the addition order of lactase according to the channel number sequence, import the enzyme addition instruction into the corresponding channel sample in sequence, collect the lactase action time setting value under the enzyme addition state, and number mark the setting value under the differential channel to obtain a channel lactase action time table; According to the channel number sequence, execute the lactase addition process, and import the required lactase volume of each channel into the corresponding sample channel as an instruction, read the number verification field before each addition operation, confirm that the channel sequence and volume value are correct, and then trigger the addition action, the addition sequence strictly follows the ascending order of the number to operate, after completing the lactase injection action of each channel, immediately record the lactase action time setting value of the corresponding channel, set T01 to 10 minutes, T02 to 12 minutes, and set different action times for part of the channels due to the difference in initial lactose concentration, which needs to be numbered and marked separately for subsequent identification and processing. Each action time value needs to be bound to the channel number and marked as a standard time value or a special setting value, each record includes channel number, lactase addition volume, action time, and whether it is a difference marker field, etc. Boolean value is used to indicate whether it is a unified reaction time, set T01 to no and T03 to yes, and arrange a channel lactase action time table.

[0029] S303: Use the action time value in the channel lactase action time table to calculate the channel temperature adjustment weight value combined with the sample reaction temperature value read in the real-time temperature control unit, combine the lactase action time and the corresponding temperature parameter according to the channel number to generate a lactose hydrolysis execution list; The channel temperature adjustment weight value uses the formula: ; Wherein, Channel temperature adjustment weight value, representing the sample reaction temperature read at the th time point, representing the average value of the sample reaction temperature at the th time point, representing the temperature perturbation compensation value corresponding to the th channel, n represents the number of channels; Formula calculation logic: symbol represents the absolute value of the deviation of each temperature sampling point from the average temperature, and the purpose is to quantify the instantaneous deviation; multiplication combines the current temperature and the perturbation compensation value to reflect the real acting temperature; the denominator term is a normalized adjustment factor under the weighting of the deviation; the overall operation logic of the formula is to obtain the overall temperature control response capability by weighted summation of each temperature deviation term, so as to avoid the interference of extreme values; The channel temperature adjustment weight value is a numerical index for measuring the influence degree of temperature fluctuation in the reaction channel on the enzyme reaction efficiency, which comprehensively considers the degree of temperature deviation from the average value at each time point and the actual temperature influence after perturbation compensation. The higher the value, the more active the temperature control response in the channel, and the stronger the regulation ability to the reaction process; Parameter meaning and calculation process: is the reaction temperature at the th time point, which is obtained by real-time temperature sampling of the actual sample in the reaction chamber through the temperature sensor; is the arithmetic mean value at the th time point, which measures the temperature stability baseline of the channel; is the sample perturbation response temperature, which reflects the temperature change compensation value caused by local perturbation (such as liquid inflow fluctuation or local heating), which can be obtained by fitting or regression of the temperature change rate trend collected by the sensor; is the total number of channel sampling points, which is a count dimensionless parameter; Parameter acquisition / quantization method and numerical example: The temperature value is obtained as follows: The temperature values of the channel sample at different time points are sampled by integrating the NTC temperature probe (0.1℃ precision), and the sampling period is set to 1 minute, a total of 5 times; as listed below: ; ; ; ; ; Average temperature : ; Temperature perturbation compensation value : By monitoring the temperature variation rate (e.g. 1 minute change rate) and liquid disturbance feedback coefficient, the following empirical fitting formula is used: ; Where, is the disturbance coefficient, the conventional experimental range is set to , which is obtained by high-frequency sampling when the sample is fed: ; ; ; ; ; Channel number : The number of sample channels in this experiment is 5, , the actual number of detection channels, Table 1: Temperature sampling and disturbance compensation data table

[0030] As shown in Table 1, the actual temperature readings of 5 sampling channels and the perturbation compensation values estimated by disturbance are listed, and the key values used for formula operation are summarized; Formula value substitution and complete calculation process: Substitute the above parameters into the formula: ; Molecule calculation: ; ; ; ; ; Molecule sum: ; Denominator calculation: ; Substitute the formula for calculation: ; The results show that the channel temperature adjustment weight value is 6.197, and the preset temperature control reaction adjustment range of this experiment is 5.0-7.5. The result shows that the temperature adjustment response ability of the current reaction channel is in the effective range, which belongs to the middle and upper response level, which can be directly used as the input parameter of the subsequent lactase action time adjustment logic; The benefit of the formula is that by introducing the disturbance compensation term with deviation weight The weighted product can truly reflect the comprehensive impact of temperature deviations at different sampling points on the overall responsiveness of the channel; compared with the traditional single-point average temperature assessment method, this method has stronger disturbance adaptability and can adapt to scenarios with slight temperature control fluctuations.

[0031] Please see Figure 5 The specific steps for obtaining the temperature control matching completion number set are as follows: S401: Call the lactase action time and reaction temperature parameters of the channel in the lactose hydrolysis execution list, identify the time node after hydrolysis is completed, import the luminescent enzyme solution into the corresponding channel sample in sequence according to the channel number, record the import time and channel number, and integrate the channel import status to generate a channel luminescent enzyme import record set. For each channel sample, the lactase action phase is identified to determine if it has been completed. Based on the recorded start injection time and action duration, the current time is calculated to determine if the reaction endpoint has been reached. Once the determination is complete, the corresponding channel's luminescent enzyme solution injection program is immediately triggered. The injection sequence strictly follows the channel number, starting from T01 and proceeding downwards. Before each injection, the action time completion node of the current channel is read. After confirming that the conditions are met, the luminescent enzyme injection action is performed. When the injection is completed, the specific time of the operation is recorded and paired with the channel number. For example, T01 is injected at 10:35, and T02 is injected at 10:38. After the injection operation is completed, each record is integrated, including the channel number, luminescent enzyme injection time, and whether the injection is completed. This information is used to determine the start conditions for subsequent temperature-controlled heating. The record table structure must clearly indicate the execution status of each channel and indicate whether the injection operation is completed. If a channel has not been injected due to unmet conditions, the status must be clearly marked as "incomplete" or "pending" in the record for subsequent supplementary operation identification and data backtracking processing, generating a channel luminescent enzyme import record set.

[0032] S402: Based on the time node information in the channel luminescent enzyme import record set, set the temperature control point and stabilization time interval of the temperature control device in the corresponding channel, determine whether the set temperature has been reached and the set stabilization time length has been maintained for each channel, mark the channels that meet the conditions as temperature regulation completed, and generate a set of temperature control matching completion numbers. The temperature control adjustment device under the corresponding channel is set with a temperature rise target and a holding time parameter, and the temperature rise starting time and the target temperature are set for each channel. The temperature rise target is set to 45°C, and the stable holding time is 8 minutes. The temperature control operation starts immediately after the channel completes the luciferase injection. By monitoring the current channel sample temperature trend, it is identified whether the target temperature is reached within a reasonable time, and whether the set holding time period is completed. T01 is injected at 10:35, the target temperature needs to be reached before 10:40 and maintained between 10:40 and 10:48. The judgment standard is whether the continuously monitored temperature is within ±0.2°C range and the duration is not less than the set value. The channels that meet the conditions are marked as "completed" in the record, and those that do not meet the conditions are marked as "not completed" and the reasons are recorded, such as "temperature rise timeout" or "temperature instability". The judgment results are summarized according to the channel number to generate a temperature control matching completion number set.

[0033] Please refer to Figure 6 The sample ATP reaction intensity classification result acquisition step is specifically: S501: Use the channel numbers in the temperature control matching completion number set to activate the luciferase system ATP reaction in the multi-channel. According to the reaction of the triggered luciferase and the substrate in the sample, the fluorescence signal intensity data in the channel is continuously recorded to generate a fluorescence intensity change data set; The corresponding channel under the corresponding channel is activated in order according to the number. In this stage, the enzyme components in the luciferase solution react with the substrate produced by the hydrolysis of lactose in the sample, and release fluorescence signals that can be detected by the photoelectric detection device. The fluorescence signal is continuously collected by the photoelectric detection component under the corresponding channel at a set frequency. Set to collect once per second, and record the current time point and the corresponding fluorescence intensity value each time. This process continues in each channel until the end of the detection time window. The monitoring period is set to 5 minutes, and a total of 300 groups of fluorescence intensity data are obtained. The collection frequency and data saving strategy need to be set uniformly to ensure stable signal collection and no missing data. The fluorescence signal record of each channel in the collection process should be bound with the number field to form a data format of three columns of channel number, time stamp and fluorescence intensity. After the channel collection is completed, the fluorescence intensity change data set is arranged.

[0034] S502: According to the fluorescence intensity change data set, the channel fluorescence signal is divided according to the time axis, the signal intensity change sequence with time is integrated into a continuous response curve, the channel number information is called to classify and arrange the curve, and the start and end time of the change section and the corresponding intensity value in the response curve are extracted to obtain the channel fluorescence response section value set; The signal data of the channel is sorted and segmented in time sequence order, and after matching a set of fluorescence intensities recorded per second with the corresponding time, the complete time sequence change track is integrated, the sequence data of the fluorescence intensity change with time is formed into a continuous response curve for each channel, and the response curve needs to be smoothed before being drawn to avoid signal acquisition errors leading to curve jumps or abnormal breakpoints. After the processing is completed, the response curve is classified and managed by calling the channel number information, and is arranged and named according to the channel number, and T01curve, T02curve, etc. are set. The obvious change section in each response curve is identified, including the stage of rapid rise or fall of fluorescence intensity, and the time period corresponding to this stage is the response section. The start time point and the end time point of the section are extracted, and the minimum value and the maximum value in the time period are recorded as the intensity range of the section. The start time of the section in the T01 curve is set to 90 seconds, the end time is set to 180 seconds, and the intensity range is set to 52 to 88. The change section extraction result of the channel is arranged into a channel fluorescence response section value set.

[0035] S503: Based on the channel fluorescence response section value set, the maximum-minimum normalization method is used to normalize the response intensity, and the normalized response amplitude is compared with the preset intensity threshold to classify the channel into high, medium and low intensity categories, and generate the sample ATP reaction intensity classification result; The intensity value and change duration in each record are extracted, and normalization processing is performed to map the response results of each channel under different intensity ranges and time scales to a unified ratio range. The normalization is processed by the maximum-minimum standardization method, that is, the maximum and minimum values in the response of all channels are taken as the unified upper and lower limits, and each channel is processed separately to normalize the intensity and time to between 0 and 1. After normalization, the response curve characteristics of each channel are classified, and the classification standard is constructed according to the normalized response amplitude and duration. Channels with intensity higher than 0.8 and duration longer than 0.7 are classified as "high intensity type", channels with intensity between 0.5 and 0.8 and duration between 0.3 and 0.7 are classified as "medium intensity type", and channels with intensity lower than 0.5 or duration shorter than 0.3 are classified as "weak response type". After classification, the normalized response result corresponding to each channel is classified into the corresponding category to generate the sample ATP reaction intensity classification result.

[0036] The above is only a specific embodiment of the present application, but the protection scope of the present application is not limited thereto. Any skilled person in the art can easily think of changes or replacements within the technical scope disclosed by the present application, which should be covered within the protection scope of the present application. Therefore, the protection scope of the present application should be subject to the protection scope of the claims.

Claims

1. A rapid method for detecting microorganisms in a fermented milk product based on ATP, characterized in that, The method comprises the following steps: S1: based on the liquid sample of the infant formula product, introducing the independent sample unit of the multi-channel liquid distribution device, sequentially collecting the sample volume reading in the sample unit, and classifying and judging with the distribution control threshold area to generate a sample distribution channel number set; S2: through the sample distribution channel number set, the corresponding channel sample is injected into the temperature control adjustment unit, the initial temperature reading and the sample lactose component reading are read, the lactose reading is compared with the lactose judgment reference range, and a lactose judgment channel label table is generated; S3: according to the lactose judgment channel label table, the channel number including lactose is extracted in sequence, the lactose reading is called and the corresponding lactase addition volume is matched, the enzyme addition sequence is configured and then the sample is introduced, according to the sample reaction temperature, through the preset temperature-time function relationship, the lactase action time setting value is determined, and a lactose hydrolysis execution list is generated; S4: calling the lactose hydrolysis execution list, introducing the luminescent enzyme solution into the corresponding channel sample according to the time node after hydrolysis is completed, setting the temperature control point and the stable time section of the temperature control adjustment device, marking the temperature adjustment completion state of each channel, and generating a temperature control matching completion number set.

2. The method for rapid detection of microorganisms based on the ATP of the fermented milk product according to claim 1, characterized in that, The sample distribution channel number set includes a channel number set, a distribution volume control number, and a sample unit number. The lactose judgment channel label table includes a lactose signal classification label, a channel lactose detection state, and a channel number annotation result. The lactose hydrolysis execution list includes lactase addition volume information, lactase action time setting value, and sample reaction temperature setting parameter. The temperature control matching completion number set includes channel temperature control completion state identification, temperature control point number, and stable time section identification.

3. The method for rapid detection of microorganisms based on the ATP of the rearing milk product according to claim 1, characterized in that, The acquisition step of the sample distribution channel number set is specifically: S101: based on the liquid sample of the infant formula product, introducing the independent sample unit of the multi-channel liquid distribution device, sequentially collecting the sample volume reading in the sample unit, calling the volume sensing component connected to the sample unit in the multi-channel liquid distribution device, acquiring the real-time volume sampling data of the channel, and generating a channel volume data set; S102: according to the channel volume data set, comparing the volume value in the sample unit with the upper and lower limit values of the volume distribution control threshold area in the sample distribution control, screening the sample unit number with the volume value in the interval range, and recombining the number and the corresponding volume value in the screening result to obtain a screening sample channel reference value list; S103: using the screening sample channel reference value list, assigning an independent volume control number to the sample unit number, pairing the number with the original number in the sample unit one by one, and binding the volume control number and the original sample number to the sample distribution channel together to generate a sample distribution channel number set.

4. The method for rapid detection of microorganisms based on the product of the rearing milk according to claim 3, characterized in that, The acquisition step of the lactose judgment channel label table is specifically: S201: Based on the sample distribution channel number set, the channel sample corresponding to the sample distribution channel number set is sequentially injected into the temperature control adjustment unit, the sample injection interface and the internal temperature control tank of the temperature control unit are called, the sample injection is sequentially completed by number control, the initial temperature value when the channel sample is injected is read, the temperature parameter data is uniformly recorded, and a sample channel temperature reading set is generated; S202: According to the channel number information in the sample channel temperature reading set, the corresponding lactose component reading in the channel sample is collected, the lactose reading is uniformly arranged under the corresponding channel number, a one-to-one correspondence structure of channel number and lactose reading is established, and a channel lactose reading value set is obtained; S203: According to the lactose reading in the channel lactose reading value set, the upper and lower limit values of the lactose judgment reference range are called, the channel corresponding lactose reading is compared with the reference range, it is judged whether there is a detectable lactose signal in the reference interval, and the judgment result is marked according to the channel number, and a channel lactose judgment mark set is generated; S204: Based on the mark state in the channel lactose judgment mark set, the number information in the sample distribution channel number set is called, the channel number and the corresponding lactose determination state are bound and combined, and are classified according to the determination result, marked as two kinds of labels of detectable and undetectable, the label information of the channel is summarized, and a lactose determination channel label table is generated.

5. The method for rapid detection of microorganisms based on the product of the rearing milk according to claim 4, characterized in that, The acquisition step of the lactose hydrolysis execution list is specifically: S301: Based on the lactose determination channel label table, the channel number marked as detectable lactose signal is extracted in sequence, the lactose reading under the corresponding channel is called, and the lactose reading is matched with the set lactase adding volume to generate a channel lactase volume value table; S302: According to the channel lactase volume value table, the adding order of lactase is arranged according to the channel number sequence, the enzyme adding instruction is sequentially introduced into the corresponding channel sample, the lactase action time setting value under the enzyme adding state is collected, and the setting value under the differential channel is numbered, marked and arranged to obtain a channel lactase action time table; S303: The action time value in the channel lactase action time table is used to calculate the channel temperature regulation weight value in combination with the sample reaction temperature value read in the real-time temperature control unit, the channel number is combined correspondingly, the lactase action time and the corresponding temperature parameter are combined, and a lactose hydrolysis execution list is generated.

6. The method for rapid detection of microorganisms based on the product of the rearing milk according to claim 5, characterized in that, The channel temperature regulation weight value is a numerical index for measuring the influence degree of temperature fluctuation in the reaction channel on the enzyme reaction efficiency.

7. The method for rapid detection of microorganisms based on the product of the rearing milk according to claim 5, characterized in that, The acquisition step of the temperature control matching complete number set is specifically: S401: The channel lactase action time and reaction temperature parameter in the lactose hydrolysis execution list are called, the time node after hydrolysis is identified, the luminous enzyme solution is sequentially introduced into the corresponding channel sample according to the channel number, the introduction time and the channel number are recorded, and the channel introduction state is integrated to generate a channel luminous enzyme introduction record set; S402: Based on the time node information in the channel luminescent enzyme import record set, set the temperature control point and stable time section of the temperature control adjustment device under the corresponding channel, judge whether the set temperature is reached and maintained for the set stable time length channel by channel, mark the channels meeting the conditions as temperature adjustment completion state, and generate a temperature control matching completion number set.

8. The method for rapid detection of microorganisms based on the ATP of the fermented milk product according to claim 7, characterized in that, The channel lactase action time is the duration required for lactase to react with lactose in the detection channel; The reaction temperature parameter is the temperature control value corresponding to the lactase action time.

9. The method for rapid detection of microorganisms based on the ATP of the rearing milk product according to claim 1, characterized in that, The method further comprises a S5 step: S5: Based on the temperature control matching completion number set, activate the ATP reaction of the multi-channel luminescent enzyme system, record the fluorescence signal intensity change in each channel, divide the fluorescence response intensity curve according to the time change, record the reaction information of the channel response section, and generate a sample ATP reaction intensity classification result; The sample ATP reaction intensity classification result includes fluorescence signal intensity time change data, response section reaction information set, and channel response intensity level label.

10. The method for rapid detection of microorganisms based on the ATP of the fermented milk product according to claim 9, characterized in that, The acquisition step of the sample ATP reaction intensity classification result is specifically: S501: Use the channel numbers in the temperature control matching completion number set to activate the ATP reaction of the multi-channel luminescent enzyme system, continuously record the fluorescence signal intensity data under the channel according to the reaction of the triggered luminescent enzyme and the substrate in the sample, and generate a fluorescence intensity change data set; S502: According to the fluorescence intensity change data set, divide the channel fluorescence signal according to the time axis, integrate the signal intensity change sequence with time into a continuous response curve, call the channel number information to classify and arrange the curve, and extract the start and end time and corresponding intensity value of the change section in the response curve to obtain a channel fluorescence response section value set; S503: Based on the channel fluorescence response section value set, use the maximum-minimum normalization method to normalize the response intensity, compare the normalized response amplitude with the preset intensity threshold, and classify the channel into high, medium and low intensity categories to generate a sample ATP reaction intensity classification result.

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