CN116355988A - Single-molecule nucleic acid detection process based on digitization - Google Patents

Single-molecule nucleic acid detection process based on digitization Download PDF

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CN116355988A
CN116355988A CN202310326913.1A CN202310326913A CN116355988A CN 116355988 A CN116355988 A CN 116355988A CN 202310326913 A CN202310326913 A CN 202310326913A CN 116355988 A CN116355988 A CN 116355988A
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袁亮
孙正道
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Jiangsu Maiyue Biotechnology Co ltd
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Abstract

The invention discloses a single-molecule nucleic acid detection process based on digitalization, which relates to the technical field of nucleic acid detection and comprises a classification planning unit, a nucleic acid extraction unit, a data acquisition unit, a nucleic acid comparison unit, a detection analysis unit and a data feedback unit. According to the invention, the nucleic acid sampling module and the nucleic acid integrating module are used for integrating the currently collected nucleic acid sample, the nucleic acid sampling module and the nucleic acid integrating module are sequentially arranged according to the time of nucleic acid collection, the single-molecule nucleic acid is divided into a plurality of nucleic acid detection units by the nucleic acid dividing module, and the single-molecule nucleic acid sample is subjected to a circulating nucleic acid amplification reaction and a hybridization reaction by using a nucleic acid amplification method such as PCR (polymerase chain reaction) according to the preset data circulating module, so that the phenomena that the detection result and the detection person are not corresponding to each other and the timeliness of detection feedback is low when the excessive single-molecule nucleic acid is detected are avoided, the calculation workload of detecting a large amount of single-molecule nucleic acid is integrated and planned, and the detection efficiency of the single-molecule nucleic acid is improved.

Description

Single-molecule nucleic acid detection process based on digitization
Technical Field
The invention relates to the technical field of nucleic acid detection, in particular to a single-molecule nucleic acid detection process based on digitalization.
Background
A single-molecule nucleic acid detection technology based on digitalization is a high-speed and long-sequence detection method with great potential, which can be used for detecting DNA, RNA, mRNA and the like, and when the method is executed, an optical system is needed to continuously monitor each polymerase which replicates template DNA and RNA molecules (molecules to be sequenced) and identify fluorescent signals related to introduction events, and thus nucleic acid base sequences of the template molecules can be read out.
However, in the conventional nucleic acid detection method, a human body fluid sample is usually collected by a nasal swab, a pharyngeal swab or the like, and a professional medical staff performs a series of processes such as extraction, detection and analysis on the nucleic acid sample to obtain a single-molecule nucleic acid detection result, but in the process of detecting a large number of nucleic acid detection samples, the detection result is not corresponding to a detector because of complicated detection procedures and easy disturbance of the collection sequence of the nucleic acid samples, so that the detection result is inevitably missed or not positive, and the timeliness of detection is reduced.
Therefore, it is necessary to provide a single-molecule nucleic acid detection process based on digitization to solve the above problems.
Disclosure of Invention
The invention aims to provide a single-molecule nucleic acid detection process based on digitization, so as to solve the problems in the background art.
In order to achieve the above purpose, the present invention provides the following technical solutions: the utility model provides a single molecule nucleic acid detection technology based on it is digital, includes classification planning unit, nucleic acid extraction unit, data acquisition unit, nucleic acid comparison unit, detects analysis unit, data feedback unit, classification planning unit's output is connected with the input of nucleic acid extraction unit, the output of nucleic acid extraction unit is connected with the input of data acquisition unit, the output of data acquisition unit is connected with the input of nucleic acid comparison unit, the output of nucleic acid comparison unit is connected with the input of detection analysis unit, the output of detection analysis unit is connected with the input of data feedback unit.
Preferably, the classification planning unit comprises a nucleic acid sampling module, a nucleic acid integration module and a nucleic acid dividing module, wherein the output end of the nucleic acid sampling module is connected with the input end of the nucleic acid integration module, and the output end of the nucleic acid integration module is connected with the input end of the nucleic acid dividing module.
Preferably, the output end of the nucleic acid dividing module is connected with the input end of the nucleic acid extracting unit.
Preferably, the data acquisition unit comprises a database module and a data circulation module, and the output end of the database module is connected with the input end of the data circulation module.
Preferably, the input end of the database module is connected with the output end of the nucleic acid extraction unit, and the output end of the data circulation module is connected with the input end of the nucleic acid comparison unit.
Preferably, the nucleic acid comparison unit comprises a data drawing module, a data calculation module, a data change module and a data generation module, wherein the output end of the data drawing module is connected with the input end of the data calculation module, the output end of the data calculation module is connected with the input end of the data change module, and the output end of the data change module is connected with the input end of the data generation module.
Preferably, the input end of the data drawing module is connected with the output end of the data acquisition unit, and the output end of the data generation module is connected with the input end of the detection analysis unit.
Preferably, the data feedback unit comprises a data sorting module, a data storage module and a data output module, wherein the output end of the data sorting module is connected with the input end of the data storage module, and the output end of the data storage module is connected with the input end of the data output module.
Preferably, the input end of the data output module is connected with the output end of the detection and analysis unit.
The invention also discloses a single-molecule nucleic acid detection process based on digitalization, which specifically comprises the following steps:
s1, during operation, a nucleic acid sample collected at present is processed through a nucleic acid sampling module contained in a classification planning unit, then the nucleic acid sample collected in a concentrated mode is integrated through a nucleic acid integration module, the nucleic acid samples are sequentially arranged according to the time of nucleic acid collection, the nucleic acid sample to be detected after being sequentially arranged is divided into a plurality of nucleic acid detection units carrying single-molecule nucleic acids through a nucleic acid dividing module, meanwhile, a plurality of single-molecule nucleic acids are extracted through a nucleic acid extracting unit, fluorescent information data corresponding to the single-molecule nucleic acids in a database are transmitted through an internal terminal processor, and the acquired nucleic acid sample is subjected to a cyclic nucleic acid amplification reaction and hybridization reaction by using a nucleic acid amplification method such as PCR (polymerase chain reaction) according to a preset data cyclic module.
S2, when single-molecule nucleic acid passes through the cyclic nucleic acid amplification reaction and the hybridization reaction, the data mapping module is used for comparing positive nucleic acid data in the database, the comparison data of the single-molecule nucleic acid is mapped into a graph, the data calculation module is used for calculating according to the graph of the single-molecule nucleic acid to obtain a single-molecule nucleic acid intensity difference value under the same amplification cycle reaction times, the data change module is used for constructing a linear change comparison relation between the single-molecule cyclic nucleic acid amplification reaction and the hybridization reaction times and the single-molecule nucleic acid intensity change difference value according to the variation difference value of the single-molecule nucleic acid intensity, and the data generation module is used for generating detection data of the single-molecule nucleic acid detection unit according to the obtained linear change comparison relation.
S3, the detection analysis unit obtains the total value of the nucleic acid concentration of the single-molecule nucleic acid in the cyclic amplification reaction according to the generated detection data of the single-molecule nucleic acid detection unit, analyzes the total value of the nucleic acid concentration of the single-molecule nucleic acid and the numerical value of the positive nucleic acid concentration in the data storage library, when the nucleic acid concentration of the single-molecule nucleic acid reaches the numerical value of the positive nucleic acid concentration, the data arrangement module arranges the detected single-molecule nucleic acid data, the data storage module stores the single-molecule nucleic acid detection data into the data storage library, the data output module transmits the detection report of the single-molecule nucleic acid to the terminal processor through the data output end, and the data display screen displays the detection report.
The invention has the technical effects and advantages that:
1. according to the single-molecule nucleic acid detection process based on the digitization, the nucleic acid sampling module and the nucleic acid integration module are used for integrating the nucleic acid samples collected at the current day, the nucleic acid samples are sequentially arranged according to the time of nucleic acid collection, the single-molecule nucleic acid is divided into a plurality of nucleic acid detection units by the nucleic acid dividing module, and the single-molecule nucleic acid samples are subjected to the cyclic nucleic acid amplification reaction and hybridization reaction by using a nucleic acid amplification method such as PCR (polymerase chain reaction) according to the preset data cyclic module, so that the phenomena that the detection result and the detection person are not corresponding to each other and the timeliness of detection feedback is low when the excessive single-molecule nucleic acid is detected are avoided, the operation workload of a large amount of single-molecule nucleic acid detection data is integrated and planned, and the detection efficiency of the single-molecule nucleic acid is improved;
2. according to the single-molecule nucleic acid detection process based on digitalization, positive nucleic acid data in a data base are compared through a data drawing module, the comparison data of single-molecule nucleic acid are drawn into a graph, and then the data calculation module calculates according to the graph of single-molecule nucleic acid to obtain a single-molecule nucleic acid intensity difference value under the same amplification cycle reaction times, so that a linear change comparison relation between single-molecule cycle nucleic acid amplification reaction and hybridization reaction times and the single-molecule nucleic acid intensity difference value is built through a data change module according to the variation difference value of single-molecule nucleic acid intensity, and the data generation module generates detection data of a single-molecule nucleic acid detection unit according to the obtained linear change comparison relation, so that the detection accuracy of a single-molecule nucleic acid detection sample is improved;
3. according to the single-molecule nucleic acid detection process based on digitization, the total nucleic acid concentration value of single-molecule nucleic acid in the cyclic amplification reaction is obtained from the detection data of the single-molecule nucleic acid detection unit, so that the total nucleic acid concentration value of single-molecule nucleic acid and the numerical value of positive nucleic acid concentration in the data storage library are analyzed, when the nucleic acid concentration of single-molecule nucleic acid reaches the numerical value of positive nucleic acid concentration, the detected single-molecule nucleic acid data can be sorted through the data sorting module, the single-molecule nucleic acid detection data are stored in the data storage library through the data storage module, meanwhile, the detection report of single-molecule nucleic acid is transmitted to the terminal processor through the data output end by the data output module, the detection report is displayed through the data display screen, and the real-time performance of the data storage update of single-molecule nucleic acid detection is greatly improved.
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FIG. 1 is a schematic diagram of a single-molecule nucleic acid detection process based on digitization.
Fig. 2 is a schematic diagram of a sort planning unit structure according to the present invention.
Fig. 3 is a schematic diagram of a data acquisition unit according to the present invention.
FIG. 4 is a schematic diagram of the structure of a nucleic acid alignment unit according to the present invention.
Fig. 5 is a schematic diagram of a data feedback unit according to the present invention.
Fig. 6 is a schematic flow diagram of a terminal processor according to the present invention.
In the figure: 1. a classification planning unit; 2. a nucleic acid extraction unit; 3. a data acquisition unit; 4. a nucleic acid comparison unit; 5. a detection analysis unit; 6. a data feedback unit; 101. a nucleic acid sampling module; 102. a nucleic acid integration module; 103. a nucleic acid partitioning module; 301. a database module; 302. a data circulation module; 401. a data drafting module; 402. a data calculation module; 403. a data change module; 404. a data generation module; 601. a data arrangement module; 602. a data storage module; 603. and a data output module.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Referring to fig. 1-4, the present invention provides a technical solution:
embodiment 1,
The utility model provides a single molecule nucleic acid detection technology based on it is digital, including classifying and planning unit 1, nucleic acid extraction unit 2, data acquisition unit 3, nucleic acid compares unit 4, detect analysis unit 5, data feedback unit 6, classifying and planning unit 1's output is connected with the input of nucleic acid extraction unit 2, the output of nucleic acid extraction unit 2 is connected with the input of data acquisition unit 3, the output of data acquisition unit 3 is connected with the input of nucleic acid compares unit 4, the output of nucleic acid compares unit 4 and detects analysis unit 5's input, detect analysis unit 5's output and data feedback unit 6's input are connected.
In order to improve the simplicity and timeliness of single-molecule nucleic acid detection, in the invention, the classification planning unit 1 comprises a nucleic acid sampling module 101, a nucleic acid integration module 102 and a nucleic acid dividing module 103, wherein the output end of the nucleic acid sampling module 101 is connected with the input end of the nucleic acid integration module 102, the output end of the nucleic acid integration module 102 is connected with the input end of the nucleic acid dividing module 103, and the detection steps of single-molecule nucleic acid can be integrated and optimized through the action of the classification planning unit 1, so that the detection timeliness of single-molecule nucleic acid is improved, and the detection efficiency of single-molecule nucleic acid is improved.
Preferably, the output of the nucleic acid dividing module 103 is connected to the input of the nucleic acid isolation unit 2 in order to accurately extract single-molecule nucleic acids.
Preferably, in order to improve the detection accuracy of the single-molecule nucleic acid, the data acquisition unit 3 includes a database module 301 and a data circulation module 302, and an output end of the database module 301 is connected to an input end of the data circulation module 302.
Preferably, to facilitate the orderly detection of the cycling nucleic acid amplification reaction and hybridization reaction, the input end of the database module 301 is connected to the output end of the nucleic acid extraction unit 2, and the output end of the data cycling module 302 is connected to the input end of the nucleic acid alignment unit 4.
As a preferred scheme, in order to better intuitively understand the single-molecule nucleic acid detection data, the nucleic acid comparing unit 4 includes a data drawing module 401, a data calculating module 402, a data changing module 403, and a data generating module 404, where an output end of the data drawing module 401 is connected to an input end of the data calculating module 402, an output end of the data calculating module 402 is connected to an input end of the data changing module 403, and an output end of the data changing module 403 is connected to an input end of the data generating module 404.
Preferably, in order to better analyze the detection data of the single-molecule nucleic acid, an input end of the data mapping module 401 is connected to an output end of the data acquisition unit 3, and an output end of the data generation module 404 is connected to an input end of the detection analysis unit 5.
As a preferred scheme, in order to update the internal information of the database in real time, more positive and negative detection data are provided, the data feedback unit 6 comprises a data sorting module 601, a data storage module 602 and a data output module 603, wherein the output end of the data sorting module 601 is connected with the input end of the data storage module 602, and the output end of the data storage module 602 is connected with the input end of the data output module 603.
Preferably, in order to conveniently feed back the data of single-molecule nucleic acid detection, the input end of the data output module 603 is connected to the output end of the detection analysis unit 5.
The invention also provides a single-molecule nucleic acid detection process based on digitalization, which comprises the novel network flow attack detection device, and further comprises the following steps:
s1, during operation, a nucleic acid sample collected at present is processed through a nucleic acid sampling module 101 contained in a classification planning unit 1, then the nucleic acid sample collected in a concentrated way is integrated through a nucleic acid integration module 102, the nucleic acid samples are sequentially arranged according to the time of nucleic acid collection, the nucleic acid sample to be detected after being sequentially arranged is divided into a plurality of nucleic acid detection units carrying single-molecule nucleic acids through a nucleic acid dividing module 103, meanwhile, a plurality of single-molecule nucleic acids are extracted through a nucleic acid extracting unit 2, fluorescent information data corresponding to the single-molecule nucleic acids in a database are transmitted through an internal terminal processor through a database module 301, and a circulating nucleic acid amplification reaction and a hybridization reaction are carried out on the collected nucleic acid sample through a nucleic acid amplification method such as PCR (polymerase chain reaction) according to a preset data circulation module 302.
S2, when single-molecule nucleic acid passes through the cyclic nucleic acid amplification reaction and the hybridization reaction, the data mapping module 401 maps the comparison data of the single-molecule nucleic acid into a graph by comparing with positive nucleic acid data in a database, the data calculation module 402 calculates according to the graph of the single-molecule nucleic acid to obtain a single-molecule nucleic acid intensity difference value under the same amplification cycle times, the data change module 403 builds a linear change comparison relation between the single-molecule cyclic nucleic acid amplification reaction and the hybridization reaction times and the single-molecule nucleic acid intensity change difference value according to the variation difference value of the single-molecule nucleic acid intensity, and the data generation module 404 generates detection data of the single-molecule nucleic acid detection unit according to the obtained linear change comparison relation.
S3, the detection analysis unit 5 obtains the total nucleic acid concentration value of the single-molecule nucleic acid in the cyclic amplification reaction according to the detection data of the generated single-molecule nucleic acid detection unit, analyzes the total nucleic acid concentration value of the single-molecule nucleic acid and the numerical value of the positive nucleic acid concentration in the data storage library, when the nucleic acid concentration of the single-molecule nucleic acid reaches the numerical value of the positive nucleic acid concentration, the data arrangement module 601 arranges the detected single-molecule nucleic acid data, the data storage module 602 stores the single-molecule nucleic acid detection data in the data storage library, the data output module 603 transmits the detection report of the single-molecule nucleic acid to the terminal processor through the data output end, and the data display screen displays the detection report.
Finally, it should be noted that: the foregoing description is only illustrative of the preferred embodiments of the present invention, and although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications may be made to the embodiments described, or equivalents may be substituted for elements thereof, and any modifications, equivalents, improvements or changes may be made without departing from the spirit and principles of the present invention.

Claims (10)

1. The single-molecule nucleic acid detection process based on digitalization comprises a classification planning unit (1), a nucleic acid extraction unit (2), a data acquisition unit (3), a nucleic acid comparison unit (4), a detection analysis unit (5) and a data feedback unit (6), and is characterized in that: the output end of the classification planning unit (1) is connected with the input end of the nucleic acid extraction unit (2), the output end of the nucleic acid extraction unit (2) is connected with the input end of the data acquisition unit (3), the output end of the data acquisition unit (3) is connected with the input end of the nucleic acid comparison unit (4), the output end of the nucleic acid comparison unit (4) is connected with the input end of the detection analysis unit (5), and the output end of the detection analysis unit (5) is connected with the input end of the data feedback unit (6).
2. The single-molecule nucleic acid detection process according to claim 1, wherein the single-molecule nucleic acid detection process is characterized in that: the classification planning unit (1) comprises a nucleic acid sampling module (101), a nucleic acid integration module (102) and a nucleic acid dividing module (103), wherein the output end of the nucleic acid sampling module (101) is connected with the input end of the nucleic acid integration module (102), and the output end of the nucleic acid integration module (102) is connected with the input end of the nucleic acid dividing module (103).
3. The single-molecule nucleic acid detection process according to claim 2, wherein: the output end of the nucleic acid dividing module (103) is connected with the input end of the nucleic acid extracting unit (2).
4. The single-molecule nucleic acid detection process according to claim 1, wherein the single-molecule nucleic acid detection process is characterized in that: the data acquisition unit (3) comprises a database module (301) and a data circulation module (302), and the output end of the database module (301) is connected with the input end of the data circulation module (302).
5. The single-molecule nucleic acid detection process according to claim 4, wherein the single-molecule nucleic acid detection process comprises the steps of: the input end of the database module (301) is connected with the output end of the nucleic acid extraction unit (2), and the output end of the data circulation module (302) is connected with the input end of the nucleic acid comparison unit (4).
6. The single-molecule nucleic acid detection process according to claim 1, wherein the single-molecule nucleic acid detection process is characterized in that: the nucleic acid comparison unit (4) comprises a data drawing module (401), a data calculation module (402), a data change module (403) and a data generation module (404), wherein the output end of the data drawing module (401) is connected with the input end of the data calculation module (402), the output end of the data calculation module (402) is connected with the input end of the data change module (403), and the output end of the data change module (403) is connected with the input end of the data generation module (404).
7. The single-molecule nucleic acid detection process according to claim 6, wherein the single-molecule nucleic acid detection process comprises the steps of: the input end of the data drawing module (401) is connected with the output end of the data acquisition unit (3), and the output end of the data generation module (404) is connected with the input end of the detection analysis unit (5).
8. The single-molecule nucleic acid detection process according to claim 1, wherein the single-molecule nucleic acid detection process is characterized in that: the data feedback unit (6) comprises a data arrangement module (601), a data storage module (602) and a data output module (603), wherein the output end of the data arrangement module (601) is connected with the input end of the data storage module (602), and the output end of the data storage module (602) is connected with the input end of the data output module (603).
9. The single-molecule nucleic acid detection process of claim 8, wherein the single-molecule nucleic acid detection process is characterized by: the input end of the data output module (603) is connected with the output end of the detection and analysis unit (5).
10. The single-molecule nucleic acid detection process according to claim 1, wherein the single-molecule nucleic acid detection process is characterized in that: the method specifically comprises the following steps:
s1, during operation, a nucleic acid sampling module (101) contained in a classification planning unit (1) is used for processing a nucleic acid sample collected in the day, then a nucleic acid integration module (102) is used for integrating the nucleic acid sample collected in a concentrated way, the nucleic acid samples are sequentially arranged according to the time of nucleic acid collection, the nucleic acid sample to be detected after the sequential arrangement is divided into a plurality of nucleic acid detection units carrying single-molecule nucleic acids through a nucleic acid dividing module (103), meanwhile, a plurality of single-molecule nucleic acids are extracted through a nucleic acid extracting unit (2), a database module (301) is used for transmitting fluorescence information data corresponding to the single-molecule nucleic acids in a database through an internal terminal processor, and a preset data circulation module (302) is used for carrying out circular nucleic acid amplification reaction and hybridization reaction on the collected nucleic acid samples by using a nucleic acid amplification method such as PCR (polymerase chain reaction);
s2, when single-molecule nucleic acid passes through the cyclic nucleic acid amplification reaction and hybridization reaction, a data mapping module (401) maps the comparison data of the single-molecule nucleic acid into a graph by comparing with positive nucleic acid data in a database, a data calculation module (402) calculates according to the graph of the single-molecule nucleic acid to obtain a single-molecule nucleic acid intensity difference value under the same amplification cycle reaction times, a data change module (403) builds a linear change comparison relation between the single-molecule cyclic nucleic acid amplification reaction and the hybridization reaction times and the single-molecule nucleic acid intensity difference value according to the variation difference value of the single-molecule nucleic acid intensity, and a data generation module (404) generates detection data of a single-molecule nucleic acid detection unit according to the obtained linear change comparison relation;
s3, a detection analysis unit (5) obtains a total nucleic acid concentration value of the single-molecule nucleic acid in the cyclic amplification reaction according to the generated detection data of the single-molecule nucleic acid detection unit, analyzes the total nucleic acid concentration value of the single-molecule nucleic acid and the numerical value of the positive nucleic acid concentration in the data storage library, when the nucleic acid concentration of the single-molecule nucleic acid reaches the numerical value of the positive nucleic acid concentration, a data arrangement module (601) arranges the detected single-molecule nucleic acid data, a data storage module (602) stores the single-molecule nucleic acid detection data in the data storage library, a data output module (603) transmits a detection report of the single-molecule nucleic acid to the terminal processor through a data output end, and a data display screen displays the detection report.
CN202310326913.1A 2023-03-30 2023-03-30 Single-molecule nucleic acid detection process based on digitization Pending CN116355988A (en)

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