阅读说明：本技术 一种基于三代测序技术的微生物检测系统 (Microorganism detection system based on third generation sequencing technology ) 是由 王浩鹏 于 2021-07-05 设计创作，主要内容包括：本发明涉及微生物检测技术领域,具体为一种基于三代测序技术的微生物检测系统,包括样品采集装置、样品振捣装置、样品检测装置以及数据显示装置,所述采集装置内设有测试液,本发明通过采用第三代测序技术对获取的微生物样品进行检测,实现了对微生物样品的检测结果的获取；通过对检测结果进行智能识别处理,实现了对检测结果的异常判断；上述方法不仅实现了对微生物样品的检测结果的获取,并且通过对检测结果进行智能识别处理,实现了对检测结果的异常判断；与传统技术相比,上述方法不需要对微生物样品进行培养,采用第三代测序技术便可实现对检测结果的获取,大大缩短了检测所耗费的时间,同时也有效地降低了对微生物检测所耗费的成本。(The invention relates to the technical field of microbial detection, in particular to a microbial detection system based on a third-generation sequencing technology, which comprises a sample collection device, a sample vibrating device, a sample detection device and a data display device, wherein a test solution is arranged in the collection device; the detection result is intelligently identified and processed, so that the abnormity judgment of the detection result is realized; the method not only realizes the acquisition of the detection result of the microorganism sample, but also realizes the abnormal judgment of the detection result by carrying out intelligent identification processing on the detection result; compared with the prior art, the method does not need to culture the microorganism sample, can realize the acquisition of the detection result by adopting a third-generation sequencing technology, greatly shortens the time consumed by detection, and simultaneously effectively reduces the cost consumed by microorganism detection.)

1. The utility model provides a little biological detection system based on third generation sequencing technology, a serial communication port, including collection system, vibration device, sample detection device (14) and data display device (7), be equipped with test solution in the collection system, test solution and sample mix, collection system fixes vibration mixing sample and test solution on the vibration device, detection device (14) adopt third generation sequencing technology right little biological sample detects, acquires the testing result, data display device (7) show the relevant information of testing result to carry out intelligent recognition to the detection and handle, judge whether the testing result exists unusually.

2. The microorganism detection system based on the third generation sequencing technology is characterized in that the sampling device comprises a sampling bottle (1), a sensor (6), tweezers and a liquid taking tube, conductors (3) are arranged on two sides of the sampling bottle (1), a sealing cap (2) is arranged at the top end of the sampling bottle (1), a sensing cap (5) is arranged at the top end of the sensor (6), a connecting groove is formed in the sensing cap (5), and a base (4) is arranged at the bottom of the sampling bottle (1).

3. The microorganism detection system based on the third generation sequencing technology, according to claim 1, characterized in that the vibration device comprises a vibrator (8) and a vibration disk (9), a vibration shaft (13) is arranged on the vibrator (8), the vibration disk (9) is installed on the vibrator (8) through the vibration shaft (13), a fixing groove (10) is arranged on the vibration disk (9), clamping blocks (11) are arranged on two sides of the fixing groove (10), and adjusting bolts (12) are arranged on two sides of the vibration disk (9) and penetrate through the vibration disk (9) to be connected with the clamping blocks (11).

4. The microorganism detection system based on the third generation sequencing technology according to claim 1, wherein the detection device (14) is a detector, a reagent tank (15), a power supply device (18) and a control unit (17) are arranged on the detector, a voltage generation device (16) is arranged in the reagent tank (15), a plurality of reagent tanks (15) are arranged, a detection cover is arranged on the reagent tank (15), a sensing rod is arranged on the detection cover, the control unit (17) comprises a detection unit, a signal processing unit, a data processing module, a processor and a storage module, the detection unit is used for collecting electric signals generated between microorganisms and the sensor (6), the signal processing unit is used for carrying out signal processing on the electric signals to obtain digital signals, and the data processing module calls pre-stored calculation parameters from the storage module through the processor, and calculating the content of the microorganisms by combining the digital signals, wherein a data transmission line is arranged between the detection device (14) and the data display device (7) for connection, a control unit is arranged in the processor, and the content displayed on the data display device (7) by the detector is controlled by the control unit.

5. The microorganism detection system based on the third generation sequencing technology of claim 1, wherein the data display device (7) comprises a display unit and an intelligent identification unit, the intelligent identification unit comprises an environment detection unit, the intelligent identification unit can also send out an abnormality early warning after determining the abnormality after judging whether the detection result is abnormal, and the environment detection unit detects pressure information in the environment information, and presets laser gas intensity, light propagation speed in the current environment medium, molecular mass of the microorganism sample and temperature in the environment information.

6. The microorganism detection system based on the third generation sequencing technology, as claimed in claim 1, wherein the test solution is a nanopore constructed from α -hemolysin, exonuclease is formed on the outer surface of one side of the nanopore, and cyclodextrin is covalently bonded to the inner surface of the nanopore to serve as a sensing medium; placing the nanopore within a lipid bilayer, wherein the salt concentration is different on both sides of the lipid bilayer; and providing preset voltage on two sides of the lipid bilayer to ensure that the exonuclease digests single-stranded DNA, a single base falls into the nanopore and temporarily interacts with cyclodextrin in the nanopore to influence the current flowing through the nanopore, and sequencing is realized according to the retention time, current and interference amplitude of each base in the cyclodextrin.

Technical Field

The invention relates to the technical field of microbial detection, in particular to a microbial detection system based on a third-generation sequencing technology.

Background

With the continuous progress of society, the detection of microorganisms in factory drainage, food and medicines is necessary;

the third-generation sequencing technology is a single-molecule sequencing technology, does not need PCR amplification during DNA sequencing, realizes independent sequencing of each DNA molecule, is also called a de novo sequencing technology, namely single-molecule real-time DNA sequencing, does not need amplification, directly performs sequencing on single-strand DNA/RNA by synthesis, degradation, nanopore and other modes, and has the characteristics of short detection time and low detection cost.

At present, the microorganism is detected by adopting a flat plate bacterial colony counting method basically; the plate colony counting method is that after a sample to be measured is properly diluted, microorganisms in the sample are fully dispersed into single cells, a certain amount of diluted sample liquid is coated on a plate, and after culture, each single cell grows and reproduces to form a colony visible to naked eyes, namely, one single colony represents one single cell in an original sample; and counting the bacterial colony number, and converting the bacterial colony number in the sample according to the dilution times and the sampling inoculation amount. The detection of microorganisms by the flat plate colony counting method takes 48 to 72 hours, so that a long detection time is consumed.

Disclosure of Invention

Technical problem to be solved

Aiming at the defects of the prior art, the invention provides a microorganism detection system which is high in detection efficiency and accurate in detection and is based on a third-generation sequencing technology.

(II) technical scheme

In order to achieve the purpose, the invention provides the following technical scheme: the utility model provides a microbial detection system based on third generation sequencing technology, includes collection system, vibrator, sample detection device and data display device, be equipped with test solution in the collection system, test solution and sample mix, collection system fixes and vibrates mixed sample and test solution on vibrator, detection device adopts third generation sequencing technology right the microbial sample detects, acquires the testing result, data display device shows the relevant information of detecting the result to carry out intelligent recognition to the detection and handle, judge whether there is the anomaly in the testing result.

In order to improve the using effect of the collecting device, the invention is improved in that the collecting device comprises a sampling bottle, a sensor, tweezers and a liquid taking pipe, wherein first conductors are arranged on two sides of the sampling bottle, a sealing cap is arranged at the top end of the sampling bottle, a sensing cap is arranged at the top end of the sensor, a connecting groove is formed in the sensing cap, and a base is arranged at the bottom of the sampling bottle.

In order to improve the mixing effect of the sample and the test solution, the invention has the improvement that the vibrating device comprises a vibrator and a vibrating disc, the vibrator is provided with a vibrating shaft, the vibrating disc is arranged on the vibrator through the vibrating shaft, the vibrating disc is provided with a fixing groove, clamping blocks are arranged on two sides of the fixing groove, and adjusting bolts are arranged on two sides of the vibrating disc and penetrate through the vibrating disc to be connected with the clamping blocks.

In order to improve the detection effect on the sample, the invention improves that the detection device is a detector, a reagent tank, a power supply device and a control unit are arranged on the detector, a voltage generation device is arranged in the reagent tank, a plurality of reagent tanks are arranged, a detection cover is arranged on the reagent tank, a sensing rod is arranged on the detection cover, the control unit comprises a detection unit, a signal processing unit, a data processing module, a processor and a storage module, the detection unit is used for collecting electric signals generated between microorganisms and the sensor, the signal processing unit is used for carrying out signal processing on the electric signals to obtain digital signals, the data processing module calls pre-stored calculation parameters from the storage module through the processor to calculate the content of the microorganisms by combining the digital signals, and a data transmission line is arranged between the detection device and the data display device for connection, the processor is internally provided with a control unit, and the control unit controls the content displayed on the data display device by the detector.

In order to improve the detection effect of the intelligent identification unit, the invention improves that the data display device comprises a display unit and an intelligent identification unit, the intelligent identification unit comprises an environment detection unit, the intelligent identification unit can also send out an abnormity early warning after determining the abnormity after judging whether the detection result is abnormal, the environment detection unit detects the pressure information in the environment information, and the laser gas intensity, the propagation speed of light in the current environment medium, the molecular mass of a microorganism sample and the temperature in the environment information are preset.

In order to improve the detection efficiency, the invention has the improvement that the test solution constructs a nanopore by using alpha-hemolysin, exonuclease is formed on the outer surface of one side of the nanopore, and cyclodextrin is covalently bonded on the inner surface of the nanopore to be used as a sensing medium; placing the nanopore within a lipid bilayer, wherein the salt concentration is different on both sides of the lipid bilayer; and providing preset voltage on two sides of the lipid bilayer to ensure that the exonuclease digests single-stranded DNA, a single base falls into the nanopore and temporarily interacts with the cyclodextrin in the nanopore to influence the current flowing through the nanopore, and sequencing is realized according to the retention time, current and interference amplitude of each base in the cyclodextrin.

(III) advantageous effects

Compared with the prior art, the invention provides a microorganism detection system based on a third-generation sequencing technology, which has the following beneficial effects:

according to the invention, the obtained microorganism sample is detected by adopting a third-generation sequencing technology, so that the detection result of the microorganism sample is obtained; the detection result is intelligently identified and processed, so that the abnormity judgment of the detection result is realized; the method not only realizes the acquisition of the detection result of the microorganism sample, but also realizes the abnormal judgment of the detection result by carrying out intelligent identification processing on the detection result; compared with the prior art, the method does not need to culture the microorganism sample, can realize the acquisition of the detection result by adopting a third-generation sequencing technology, greatly shortens the time consumed by detection, and simultaneously effectively reduces the cost consumed by microorganism detection.

Drawings

FIG. 1 is a schematic view of the structure of the collecting device of the present invention;

FIG. 2 is a schematic structural view of the vibrating apparatus of the present invention;

FIG. 3 is a schematic top view of the inspection apparatus of the present invention;

FIG. 4 is a schematic view of a connection structure between a detection device and a data display device according to the present invention;

in the figure: 1. a sampling bottle; 2. a sealing cap; 3. a conductor; 4. a base; 5. a sensing cap; 6. a sensor; 7. a data display device; 8. a vibrator; 9. a vibrating pan; 10. fixing grooves; 11. a clamping block; 12. Adjusting the bolt; 13. a vibration shaft; 14. a detection device; 15. a reagent tank; 16. a voltage generating device; 17. a control unit; 18. a power supply device;

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Referring to fig. 1-4, the invention is a microorganism detection system based on a third generation sequencing technology, comprising a collection device, a vibrating device, a sample detection device 14 and a data display device 7, wherein a test solution is arranged in the collection device, the test solution is mixed with a sample, the collection device is fixed on the vibrating device and vibrates the mixed sample and the test solution, the detection device 14 adopts a third generation sequencing technology to detect the microorganism sample to obtain a detection result, the data display device 7 displays relevant information of the detection result, and carries out intelligent identification processing on the detection to judge whether the detection result is abnormal.

In the embodiment, the sampling device comprises a sampling bottle 1, a sensor 6, tweezers and a liquid taking tube, conductors 3 are arranged on two sides of the sampling bottle 1, the top end of the sampling bottle 1 is provided with a sealing cap 2, the top end of the sensor 6 is provided with a sensing cap 5, the sensing cap 5 is provided with a connecting groove, the bottom of the sampling bottle 1 is provided with a base 4, clamping a solid sample into the sampling bottle 1 by using tweezers to be mixed with a test solution, absorbing the liquid sample by using a liquid taking tube, dropping the liquid sample into the sampling bottle 1 to be mixed with the test solution, the sampling bottle 1 is sealed by the sealing cap 2, then the sampling bottle 1 is shaken to uniformly mix the test solution and the sample, then the sealing cap 2 is disassembled to install the sensor 6 in the sampling bottle 1 to be contacted with the mixed solution, the sampling bottle 1 is sealed by a sensing cap 5 and connected to a detection device 14, which receives a voltage via conductors 3 on both sides of the sampling bottle 1.

In this embodiment, the device of vibrating includes vibrator 8 and vibration dish 9, be equipped with vibration axle 13 on the vibrator 8, vibration dish 9 is installed on vibrator 8 through vibration axle 13, be equipped with fixed slot 10 on the vibration dish 9, fixed slot 10 both sides are equipped with clamp splice 11, vibration dish 9 both sides are equipped with adjusting bolt 12 and run through vibration dish 9 and are connected with clamp splice 11, put into fixed slot 10 with sampling bottle 1 and make clamp splice 11 carry base 4 through rotating adjusting bolt 12, prevent through base 4 that clamp splice 11's clamp power direct action is on sampling bottle 1, cause the damage to sampling bottle 1 under the too big condition of clamp power, sampling bottle 1 is started vibrator 8 after being fixed and is driven the vibration dish vibration to make the test solution in sampling bottle 1 mix with the sample is quick and be in the same place.

In this embodiment, the detection device 14 is a detector, the detector is provided with a reagent tank 15, a power supply device 18 and a control unit 17, the reagent tank 15 is internally provided with a plurality of voltage generation devices 16, the reagent tank 15 is provided with a detection cover, the detection cover is provided with a sensing rod, the control unit 17 comprises a detection unit, a signal processing unit, a data processing module, a processor and a storage module, the detection unit is used for collecting telecommunication numbers generated between microorganisms and the sensor 6, the signal processing unit is used for processing the electric signals to obtain digital signals, the data processing module calls pre-stored calculation parameters from the storage module through the processor to calculate the content of the microorganisms by combining the digital signals, and a data transmission line is arranged between the detection device 14 and the data display device 7 for connection, the processor is internally provided with a control unit, the control unit controls the content displayed on the data display device 7 by the detector, the detection device 14 transmits voltage to the conductor 3 through the voltage generation device 16, and the sensing rod is connected with the sensing cap 5 to obtain an electric signal so as to improve the detection effect of the sample.

In this embodiment, the data display device 7 includes a display unit and an intelligent recognition unit, the intelligent recognition unit includes an environment detection unit, the intelligent recognition unit can send an abnormal warning after determining whether the detection result is abnormal or not, the environment detection unit detects pressure information in the environment information, presets laser gas intensity, propagation speed of light in the current environment medium, molecular mass of the microbial sample and temperature in the environment information, detects information data of the detection environment changed when the detection sample is obtained through the environment detection unit, judges a difference between the total amount of microbes in the adjusted detection result and the detection structure through the combination of the information data of the environment and the sample adjusted according to the historical detection data through the intelligent recognition unit, if the difference is too large, the detection result is abnormal, and detecting and sending out an abnormal early warning.

In the embodiment, the test solution uses alpha-hemolysin to construct a nanopore, exonuclease is formed on the outer surface of one side of the nanopore, and cyclodextrin is covalently bonded on the inner surface of the nanopore to serve as a sensing medium; placing the nanopore within a lipid bilayer, wherein the salt concentration is different on both sides of the lipid bilayer; and preset voltage is provided on two sides of the lipid bilayer, so that single-stranded DNA is digested by exonuclease, a single base falls into the nanopore and temporarily interacts with cyclodextrin in the nanopore, the current flowing through the nanopore is influenced, and sequencing is realized according to the retention time, current and interference amplitude of each base in the cyclodextrin, so that the detection efficiency is improved.

To sum up, the microorganism detection system based on the third generation sequencing technology clamps a solid sample into a sampling bottle 1 through tweezers to be mixed with a test solution when in use, a liquid sample is absorbed through a liquid taking tube and then is dripped into the sampling bottle 1 to be mixed with the test solution, the sampling bottle 1 is sealed through a sealing cap 2 and then is shaken to enable the test solution to be uniformly mixed with the sample, then the sealing cap 2 is disassembled to install a sensor 6 into the sampling bottle 1 to be contacted with a mixed solution, the sampling bottle 1 is sealed through a sensing cap 5 and is connected with a sample detection device 14, a conductor 3 on two sides of the sampling bottle 1 receives voltage, the sampling bottle 1 is placed into a fixed groove 10, a clamping block 11 clamps a base 4 through rotating an adjusting bolt 12, the clamping force of the clamping block 11 is prevented from directly acting on the sampling bottle 1 through the base 4, and the sampling bottle 1 is damaged under the condition of overlarge clamping force, after the sampling bottle 1 is fixed, a vibrator 8 is started to drive a vibration disc to vibrate, so that the test liquid in the sampling bottle 1 is mixed with a sample quickly, then the sampling bottle 1 is placed in a reagent tank 15, a detection device 14 transmits voltage to a conductor 3 through a voltage generation device 16, a sensing rod is connected with a sensing cap 5 to obtain an electric signal detected by a sensor 6, a signal processing unit is used for processing the electric signal to obtain a digital signal, a data processing module calls a prestored calculation parameter from a storage module through a processor to calculate the content of the microorganism by combining the digital signal, the content displayed on a data display device 7 by a control unit control detector, information data of environment change is detected when the sample is detected and obtained through an environment detection unit, and the intelligent identification unit judges the detection after adjustment according to the combination of the information data of the environment and the sample according to historical detection data If the difference between the total quantity of the microorganisms in the result and the detection structure is too large, the detection result is abnormal, and the detection is carried out to send out an abnormal early warning; the detection result is intelligently identified and processed, so that the abnormity judgment of the detection result is realized; the method not only realizes the acquisition of the detection result of the microorganism sample, but also realizes the abnormal judgment of the detection result by carrying out intelligent identification processing on the detection result; compared with the prior art, the method does not need to culture a microorganism sample, can realize the acquisition of the detection result by adopting a third-generation sequencing technology, greatly shortens the time consumed by detection, and simultaneously effectively reduces the cost consumed by microorganism detection.

Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.