CN116987585A - Oral cavity flora analysis device and method based on high-throughput sequencing - Google Patents
Oral cavity flora analysis device and method based on high-throughput sequencing Download PDFInfo
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- 229920000742 Cotton Polymers 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000011120 plywood Substances 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 238000003794 Gram staining Methods 0.000 description 1
- 241000761557 Lamina Species 0.000 description 1
- 208000025157 Oral disease Diseases 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
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Abstract
The invention discloses an oral cavity flora analysis device and method based on high-flux sequencing, and relates to the technical field of oral cavity flora measurement and analysis, wherein the oral cavity flora analysis device comprises an oral cavity saliva sampling device, a group of saliva sampling plates, an automatic high-flux colony selection instrument, a group of single-colony inoculation plates, a single-colony high-flux growth monitoring system and an automatic controller; according to the saliva sampling device based on the dental floss structure, the sampling flow is greatly simplified, the subsequent automatic detection flow is also facilitated by the saliva sampling plate, the high-flux growth monitoring system is adopted to perform high-flux detection, the image acquisition and the image processing can be simultaneously performed on 12 culture dishes, the image analysis on 2400 colonies is realized, the detection requirement on a plurality of single colonies in the oral medicine detection is met, the efficiency is higher, and the colony detection data is more detailed and accurate.
Description
Technical Field
The invention relates to the technical field of oral cavity flora determination and analysis, in particular to an oral cavity flora analysis device and method based on high-throughput sequencing.
Background
At present, people are more and more concerned about oral health, and healthy oral flora is balanced, and once the flora is unbalanced, oral diseases can occur, so that the oral health of patients is seriously affected.
At present, the traditional oral cavity microecological flora detection steps mainly comprise:
firstly, collecting saliva; secondly, tabletting saliva; thirdly, gram staining is carried out; fourth, the number of gram negative, positive cocci or bacilli was observed microscopically.
By observing and counting the flora, the oral flora can be quantitatively detected.
The traditional detection mode has low saliva collection efficiency, few detectable flora types and low detection efficiency, and cannot meet a plurality of detection requirements in a large hospital every day.
Disclosure of Invention
In order to solve the problem of efficiently detecting oral colonies, the invention provides an oral flora analysis device and an oral flora analysis method based on high-throughput sequencing. The following technical scheme is adopted:
the oral cavity flora analysis device based on high-throughput sequencing comprises an oral cavity saliva sampling device, a group of saliva sampling plates, an automatic high-throughput colony selector, a group of single-colony inoculation plates, a single-colony high-throughput growth monitoring system and an automatic controller;
one side of the saliva sampling plate is detachably connected to the oral saliva sampling device, and when the oral saliva sampling device is placed in an oral cavity, the oral saliva sampling device guides saliva in the oral cavity to the saliva sampling plate, the automatic high-flux colony selector scans a colony high-flux imaging picture on the saliva sampling plate and analyzes the picture, a selecting needle of the automatic high-flux colony selector completes colony selection on the saliva sampling plate according to analysis results and colony requirements to be selected according to parameter setting, the selected colony sample is placed on a single colony inoculation plate, then the single colony inoculation plate is placed in a culture dish of a single colony high-flux growth monitoring system for culture, a high-flux imaging monitoring module of the single colony high-flux growth monitoring system scans and monitors growth conditions of single colonies in the culture dish in real time, and records and monitors output of the results, and the automatic controller is respectively in communication connection with the automatic high-flux colony selector and the single colony high-flux growth monitoring system.
By adopting the technical scheme, saliva in the oral cavity is sampled differently from traditional cotton swab sampling, saliva can be directly and uniformly distributed on the saliva sampling plate, and the problem that uncontrollable pollution caused by manual operation in more links affects the detection result is avoided;
the automatic high-flux colony picking instrument is responsible for picking each colony on the saliva sampling plate, a high-flux scanning module of the automatic high-flux colony picking instrument can carry out high-flux imaging on the colony on the saliva sampling plate, generally, tens of picking needles are arranged, picking of different colonies can be respectively completed according to the arrangement, the picked single colony is inoculated on the single colony inoculation plate, then the single colony inoculation plate is placed in a culture dish of the single colony high-flux growth monitoring system, a bacterial colony imaging instrument connected with an automatic controller and arranged in a high-flux growth monitoring system in a constant temperature system is used, bacterial liquid in the culture dish in a fixing plate mounting hole of the bacterial colony imaging instrument is scanned in real time according to program requirements and set parameters, a bacterial liquid image is generated, the bacterial growth monitoring processing system is used for collecting and processing images generated by the bacterial colony imaging instrument, the change condition of the size of the community is tracked, so as to determine the occurrence of the community, the growth curve graph, the frequency chart of the growth, the death graph, the heat map and the like are further obtained, the monitoring period is completed, and all data are imported into the automatic controller for storage and examination; the high-flux growth monitoring system adopts two colony imagers of one system, can collect images and process images of all culture dishes simultaneously, realizes image analysis of 2400 colonies, meets the detection requirement of a plurality of single colonies in the oral medical detection, has higher efficiency, and ensures more detailed and accurate colony detection data.
Optionally, the automatic high-flux colony picking device further comprises a tray device and a manipulator, wherein the tray device and the manipulator are respectively arranged on one side of the automatic high-flux colony picking device, the tray device is used for transferring saliva sampling plates, the manipulator is used for grabbing and placing the saliva sampling plates on the tray device on a colony picking station of the automatic high-flux colony picking device, and the automatic controller controls the executing action of the manipulator.
Through adopting above-mentioned technical scheme, tray device can realize keeping in polylith saliva sampling board, for example in an oral cavity colony detects branch of academic or vocational study indoor, and the inspector can be in proper order with the saliva sampling board of many wait to examine personnel carry out the label after keeping in on tray device, and follow-up manipulator carries out single colony inoculation with the saliva sampling board that corresponds again under automatic controller's control.
Optionally, the tray device includes tray frame and automatic apron subassembly, the tray frame sets up in one side of automatic high flux bacterial colony selection appearance, and the tray frame interval is equipped with a plurality of laminas, automatic apron subassembly includes contact switch, singlechip, motor and apron, the plywood upper surface is equipped with the switch groove, contact switch sets up in switch groove department to with singlechip control connection, the power shaft of motor links to each other with one side transmission of apron, singlechip control motor's switch, when contact switch is sheltered from by saliva sampling board, singlechip control motor starts, and the drive apron covers saliva sampling board upper surface, automatic controller and singlechip communication connection.
Through adopting above-mentioned technical scheme, can set up automatic apron subassembly, when saliva sampling board contacted the plywood upper surface and triggered contact switch, contact switch sent the switching value signal for the singlechip, and the singlechip control motor starts, and the power shaft drive apron lid of motor is at saliva sampling board upper surface, avoids the bacterial colony in the air to pollute saliva sampling board.
Optionally, oral cavity saliva sampling device includes facing, lining cloth, a set of siphon connecting strip and sample cardboard, the lining cloth is laid on the facing, and when the facing was used, the saliva was collected to lining cloth direct contact tooth, and the one end of a set of siphon connecting strip is connected respectively in the inboard of lining cloth, the sample cardboard includes frame board and gauze face, the other end of a set of siphon connecting strip is connected respectively all around to the frame board to be located the interior arc intermediate position of facing, the frame board is equipped with the inside and outside channel hole that switches on of frame, the gauze face sets up in the inboard of frame board, and absorbs the saliva of siphon connecting strip guide through the channel hole, saliva sampling board lock dress is in frame board department, and the sample face contacts the gauze face, with saliva equipartition on the sample face of saliva sampling board.
Through adopting above-mentioned technical scheme, in saliva sampling, can put into the oral cavity with the facing, the tooth is tight bites, and saliva on the tooth will follow the lining cloth to siphon connecting strip this moment, finally distributes full yarn cloth cover, and whole sampling time is better 2-3 minutes, and when finally detaining saliva sampling board dress in frame board department, the sampling surface supports yarn cloth cover, and saliva on the yarn cloth cover will equipartition on the sampling surface of saliva sampling board.
The non-uniformity of sampling of the cotton swab is avoided, samples are required to be manufactured later, time is greatly reduced, and a subsequent automatic detection link is facilitated.
Optionally, the lining cloth, the siphon connecting strip and the gauze surface are all made of medical polyester fiber cloth.
By adopting the technical scheme, the medical polyester fiber cloth is high in sanitation level and has good water absorption property, and saliva in the oral cavity can be collected well.
Optionally, a transferring manipulator is arranged between the automatic high-throughput colony picking instrument and the single-colony high-throughput growth monitoring system, the transferring manipulator is used for transferring the single-colony inoculating plate into a culture dish of the single-colony high-throughput growth monitoring system, and the automatic controller controls the executing action of the transferring manipulator.
Through adopting above-mentioned technical scheme, set up the transportation manipulator between automatic high-flux bacterial colony selection appearance and single bacterial colony high-flux growth monitoring system, can realize the transportation of single bacterial colony inoculation board automatically under the control of automatic control ware, promote detection efficiency, avoid artificial operation's mistake and to the pollution of single bacterial colony inoculation board.
Optionally, the automation controller is an industrial personal computer.
By adopting the technical scheme, the industrial control computer can realize industrial automation control and can also store, display and output detection data.
Alternatively, the automated high throughput colony picker employs a RapidPick automatic clone colony picker.
By adopting the technical scheme, the rapid pick automatic clone colony picker has small occupied area, saves the space of a detection laboratory, and is easy to install in an anaerobic chamber and a biosafety cover. Colony picking rate can reach 2500 per hour. The open physical layout can be conveniently integrated with other instruments such as incubators, media dispensers, manipulators, etc.
The oral cavity flora analysis method based on high-throughput sequencing comprises the following specific steps:
step 1, saliva in the oral cavity is sampled, a dental socket is placed in the oral cavity, teeth bite into the dental socket and are kept for 2-3 minutes, the gauze surface is fully stained with saliva, the dental socket is taken out, a saliva sampling plate is clamped at a frame plate, the sampling surface of the saliva sampling plate is uniformly stained with saliva, then a label attached to the saliva sampling plate is placed on a layer plate of a tray device, and an automatic cover plate assembly starts to cover the saliva sampling plate;
step 2, bacterial colony selection, wherein an automatic controller controls a manipulator to grasp a designated saliva sampling plate, when a gripper of the manipulator reaches the position of the designated saliva sampling plate, the automatic controller controls an automatic cover plate assembly to be opened, the manipulator finishes grasping, the saliva sampling plate is placed on a bacterial colony selection station of an automatic high-flux bacterial colony selector, and the automatic high-flux bacterial colony selector respectively collects bacterial groups needing to be sampled and analyzed on the saliva sampling plate to single bacterial colony inoculation plates according to set parameters;
step 3, colony culture, wherein an automatic controller controls a transfer manipulator to transfer a single colony inoculation plate into a culture dish of a single colony high-flux growth monitoring system for culture;
and 4, detecting the bacterial colony, scanning and monitoring the growth condition of the single bacterial colony in the culture dish in real time by a single bacterial colony high-flux growth monitoring system, and outputting the recorded monitoring result to an automatic controller.
In summary, the present invention includes at least one of the following beneficial technical effects:
the invention provides an oral cavity flora analysis device and method based on high-flux sequencing, which adopts a saliva sampling device based on a dental floss structure, so that the sampling flow is greatly simplified, the saliva sampling plate is also convenient for the subsequent automatic detection flow, a high-flux growth monitoring system is adopted to perform high-flux detection, 12 culture dishes can be subjected to image acquisition and image processing at the same time, 2400 colonies are subjected to image analysis, the detection requirement of the oral medicine detection on a plurality of single colonies is met, the efficiency is higher, and the colony detection data is more detailed and accurate.
Drawings
FIG. 1 is a schematic diagram of the principle of component connection of the oral cavity flora analyzer based on high throughput sequencing of the present invention;
FIG. 2 is a schematic diagram of the oral saliva sampling device of the oral flora analyzer based on high throughput sequencing;
FIG. 3 is a schematic diagram of the tray device of the oral cavity flora analysis device based on high-throughput sequencing;
FIG. 4 is a schematic diagram of the electrical device connection principle of the oral cavity flora analyzer based on high throughput sequencing of the present invention.
Reference numerals illustrate: 1. an oral saliva sampling device; 11. a tooth socket; 12. lining cloth; 13. a siphon connection bar; 14. a sampling clamping plate; 141. a frame plate; 142. yarn cloth cover; 2. saliva sampling plate; 3. an automated high-throughput colony picker; 4. inoculating single colony to the seed plate; 5. a tray device; 51. a tray frame; 511. a laminate; 52. a contact switch; 53. a single chip microcomputer; 54. a motor; 55. a cover plate; 6. a manipulator; 7. a transfer manipulator; 8. a single colony high-throughput growth monitoring system; 10. an automated controller.
Detailed Description
The present invention will be described in further detail with reference to the accompanying drawings.
The embodiment of the invention discloses an oral cavity flora analysis device and method based on high-throughput sequencing.
Referring to fig. 1-4, a high throughput sequencing-based oral flora analysis device comprises an oral saliva sampling device 1, a group of saliva sampling plates 2, an automatic high throughput colony selector 3, a group of single colony inoculation plates 4, a single colony high throughput growth monitoring system 8 and an automatic controller 10;
one side of the saliva sampling plate 2 is detachably connected to the oral saliva sampling device 1, and when the oral saliva sampling device 1 is placed in an oral cavity, the oral saliva sampling device 1 introduces oral saliva into the saliva sampling plate 2, the automatic high-flux colony selector 3 scans colony high-flux imaging pictures on the saliva sampling plate 2 and analyzes the pictures, a selecting needle of the automatic high-flux colony selector 3 completes colony selection on the saliva sampling plate 2 according to analysis results and colony requirements required to be selected according to parameter settings, the selected colony samples are placed on the single colony inoculation plate 4, then the single colony inoculation plate 4 is placed in a culture dish of the single colony high-flux growth monitoring system 8 for culture, a high-flux imaging monitoring module of the single colony high-flux growth monitoring system 8 scans and monitors growth conditions of single colonies in the culture dish in real time, and monitoring result output is recorded, and the automatic controller 10 is respectively connected with the automatic high-flux selector 3 and the single colony high-flux growth monitoring system 8 in a communication manner.
The saliva sampling in the oral cavity is different from the traditional cotton swab sampling, saliva can be directly and uniformly distributed on the saliva sampling plate 2, and the problem that uncontrollable pollution caused by manual operation in more links affects the detection result is avoided;
the automatic high-flux colony picking instrument 3 is responsible for picking each colony on the saliva sampling plate 2, a high-flux scanning module of the automatic high-flux colony picking instrument 3 can conduct high-flux imaging on the colony on the saliva sampling plate 2, generally, tens of picking needles are arranged, picking of different colonies can be completed according to the arrangement, the picked single colony is inoculated to the single colony inoculation plate 4, the single colony inoculation plate 4 is placed in a culture dish of the single colony high-flux growth monitoring system 8, a bacterial colony imaging instrument connected with the automatic controller 10 and arranged in the high-flux growth monitoring system in a constant temperature system is used for scanning bacterial liquid in the culture dish in a fixing plate mounting hole of the bacterial colony imaging instrument according to program requirements and set parameters in real time to generate a bacterial liquid image, the bacterial growth monitoring processing system is used for collecting and processing images generated by the bacterial colony imaging instrument, the change condition of the size of the colony is tracked to determine the occurrence, growth, time points of the colony and death are further obtained, a growth frequency chart, death, a heat chart and the like are completed, and all data are imported into the automatic controller for storage and examination of the automatic controller 10; the high-flux growth monitoring system adopts two colony imagers of one system, can collect images and process images of 12 culture dishes simultaneously, realizes image analysis of 2400 colonies, meets the detection requirement of a plurality of single colonies in the oral medical detection, has higher efficiency, and ensures more detailed and accurate colony detection data.
The automatic high-flux colony picking device further comprises a tray device 5 and a manipulator 6, wherein the tray device 5 and the manipulator 6 are respectively arranged on one side of the automatic high-flux colony picking device 3, the tray device 5 is used for transshipping the saliva sampling plate 2, the manipulator 6 is used for grabbing and placing the saliva sampling plate 2 on the tray device 5 on a colony picking station of the automatic high-flux colony picking device 3, and the automatic controller 10 controls execution actions of the manipulator 6.
The tray device 5 can temporarily store a plurality of saliva sampling plates 2, for example, in an oral cavity colony detection department, a detector can sequentially label the saliva sampling plates 2 of a plurality of people to be detected and temporarily store the saliva sampling plates on the tray device 5, and the subsequent manipulator 6 performs single colony inoculation on the corresponding saliva sampling plates 2 under the control of the automatic controller 10.
The tray device 5 comprises a tray frame 51 and an automatic cover plate assembly, wherein the tray frame 51 is arranged on one side of the automatic high-flux colony picking instrument 3, a plurality of laminates 511 are arranged at intervals on the tray frame 51, the automatic cover plate assembly comprises a contact switch 52, a single-chip microcomputer 53, a motor 54 and a cover plate 55, a switch groove is formed in the upper surface of each laminate 511, the contact switch 52 is arranged at the switch groove and is in control connection with the single-chip microcomputer 53, a power shaft of the motor 54 is in transmission connection with one side of the cover plate 55, the single-chip microcomputer 53 controls the motor 54 to be switched on, when the contact switch 52 is shielded by the saliva sampling plate 2, the single-chip microcomputer 53 controls the motor 54 to be started, the cover plate 55 is driven to cover the upper surface of the saliva sampling plate 2, and the automatic controller 10 is in communication connection with the single-chip microcomputer 53.
Because it is not known how long the saliva sampling plate 2 will exist on the laminate 511 of the tray device 5, an automatic cover plate assembly can be provided, when the saliva sampling plate 2 contacts the upper surface of the laminate 511 and triggers the contact switch 52, the contact switch 52 sends a switching value signal to the singlechip 53, the singlechip 53 controls the motor 54 to start, and the power shaft of the motor 54 drives the cover plate 55 to cover the upper surface of the saliva sampling plate 2, so that bacterial colonies in the air are prevented from polluting the saliva sampling plate 2.
The oral saliva sampling device 1 comprises a tooth socket 11, an interlining 12, a group of siphon connecting strips 13 and a sampling clamping plate 14, wherein the interlining 12 is paved on the tooth socket 11, when the tooth socket 11 is used, the interlining 12 is in direct contact with teeth to collect saliva, one ends of the group of siphon connecting strips 13 are respectively connected to the inner side of the interlining 12, the sampling clamping plate 14 comprises a frame plate 141 and a yarn cloth surface 142, the periphery of the frame plate 141 is respectively connected with the other ends of the group of siphon connecting strips 13 and is positioned at the middle position of an inner arc of the tooth socket 11, the frame plate 141 is provided with a channel hole which is communicated with the inside and the outside of the frame, the yarn cloth surface 142 is arranged on the inner side of the frame plate 141 and absorbs saliva guided by the siphon connecting strips 13 through the channel hole, and when the saliva sampling plate 2 is buckled on the frame plate 141, the sampling surface contacts the yarn cloth surface 142 to uniformly distribute saliva on the sampling surface of the saliva sampling plate 2.
In saliva sampling, the dental socket 11 can be placed in the oral cavity, teeth are tightly bitten, at this time saliva on the teeth is enabled to go from the lining cloth 12 to the siphon connecting strip 13 and finally to be fully distributed on the gauze surface 142, the whole sampling time is preferably 2-3 minutes, and finally, when the saliva sampling plate 2 is buckled at the frame plate 141, the sampling surface is propped against the gauze surface 142, and saliva on the gauze surface 142 is evenly distributed on the sampling surface of the saliva sampling plate 2.
The non-uniformity of sampling of the cotton swab is avoided, samples are required to be manufactured later, time is greatly reduced, and a subsequent automatic detection link is facilitated.
The lining cloth 12, the siphon connecting strip 13 and the gauze surface 142 are all made of medical polyester fiber cloth.
The medical polyester fiber cloth material has high sanitary level and good water absorption property, and can collect saliva in the oral cavity well.
A transfer manipulator 7 is arranged between the automatic high-throughput colony picking instrument 3 and the single-colony high-throughput growth monitoring system 8, the transfer manipulator 7 is used for transferring the single-colony inoculation plates 4 into a culture dish of the single-colony high-throughput growth monitoring system 8, and an automatic controller 10 controls the execution action of the transfer manipulator 7.
The transfer manipulator 7 is arranged between the automatic high-flux colony selection instrument 3 and the single-colony high-flux growth monitoring system 8, and can automatically transfer the single-colony inoculation plates 4 under the control of the automatic controller 10, so that the detection efficiency is improved, and the errors of manual operation and the pollution to the single-colony inoculation plates 4 are avoided.
The automation controller 10 is an industrial personal computer.
The industrial control computer can realize industrial automation control and can also store, display and output detection data.
Automated high throughput colony picker 3 employs a RapidPick automatic clone colony picker.
The rapid Pick automatic clone colony picking instrument has small occupied area, saves the space of a detection laboratory, and is easy to install in an anaerobic room and a biosafety cover. Colony picking rate can reach 2500 per hour. The open physical layout can be conveniently integrated with other instruments such as incubators, media dispensers, manipulators, etc.
The oral cavity flora analysis method based on high-throughput sequencing comprises the following specific steps:
step 1, oral saliva sampling, namely placing a dental socket 11 in an oral cavity, clenching teeth into the dental socket 11, keeping for 2-3 minutes, enabling a gauze surface 142 to be full of saliva, taking out the dental socket 11, clamping a saliva sampling plate 2 at a frame plate 141, uniformly staining saliva on the sampling surface of the saliva sampling plate 2, placing a label attached to the saliva sampling plate 2 on a layer plate 511 of a tray device 5, and enabling an automatic cover plate assembly to cover the saliva sampling plate 2;
step 2, colony picking, wherein an automatic controller 10 controls a manipulator 6 to grasp a designated saliva sampling plate 2, when the gripper of the manipulator 6 reaches the position of the designated saliva sampling plate 2, the automatic controller 10 controls an automatic cover plate assembly to be opened, the manipulator 6 finishes grabbing, the saliva sampling plate 2 is placed on a colony picking station of an automatic high-flux colony picking instrument 3, and the automatic high-flux colony picking instrument 3 respectively collects bacterial groups needing to be sampled and analyzed on the saliva sampling plate 2 to single colony inoculation plates 4 according to set parameters;
step 3, colony culture, wherein an automatic controller 10 controls a transfer manipulator 7 to transfer a single colony inoculation plate 4 into a culture dish of a single colony high-flux growth monitoring system 8 for culture;
and 4, colony detection, wherein the single-colony high-flux growth monitoring system 8 scans and monitors the growth condition of single colonies in the culture dish in real time, and outputs the recorded monitoring result to the automatic controller 10.
The invention discloses an oral cavity flora analysis device and method based on high-throughput sequencing, and a specific implementation principle thereof:
under the application scene in a certain oral colony detection laboratory, the inspection staff uses disposable oral saliva sampling device 1 to sample, place tooth socket 11 in waiting to examine personnel's oral cavity, wait to examine personnel and cleft tooth socket 11 with the tooth, keep 2 minutes, make gauze face 142 be stained with saliva, take out tooth socket 11, the saliva sampling board 2 cartridge that will make the mark is in frame board 141 department, the sampling face of saliva sampling board 2 evenly is stained with saliva, paste the label with saliva sampling board 2 and place on the plywood 511 of tray device 5 again, automatic apron subassembly starts to cover saliva sampling board 2, then can carry out the saliva sampling of next waiting to examine personnel, the sampling step is the same.
The industrial personal computer controls the manipulator 6 to grasp the appointed saliva sampling plate 2, when the gripper of the manipulator 6 reaches the position of the appointed saliva sampling plate 2, the industrial personal computer controls the automatic cover plate component to be opened, the manipulator 6 finishes grasping and places the saliva sampling plate 2 on a colony selecting station of the automatic high-flux colony selecting instrument 3, the automatic high-flux colony selecting instrument 3 acquires and inoculates a single colony on the single colony inoculation plate 4 through a sampling needle of the automatic high-flux colony selecting instrument 3 according to set parameters including the types of colonies needing to be sampled, position coordinates of the colonies on the saliva sampling plate 2 and the like obtained through analysis of a high-flux imaging picture;
the single-colony high-flux growth monitoring system 8 comprises a constant temperature and humidity incubator and a high-flux colony imager, and after the steps are finished, the industrial personal computer controls the transfer manipulator 7 to transfer the single-colony inoculation plates 4 into a culture dish of the constant temperature and humidity incubator of the single-colony high-flux growth monitoring system 8 for culture;
the high-flux colony imager can scan and monitor the growth condition of single colonies in the culture dish in real time, and output the recorded monitoring result to the industrial personal computer, and finally the industrial personal computer can inquire and output the detection result.
The above embodiments are not intended to limit the scope of the present invention, and therefore: all equivalent changes in structure, shape and principle of the invention should be covered in the scope of protection of the invention.
Claims (9)
1. Oral cavity flora analytical equipment based on high throughput sequencing, its characterized in that: the automatic saliva sampling device comprises an oral saliva sampling device (1), a group of saliva sampling plates (2), an automatic high-throughput colony selector (3), a group of single-colony inoculation plates (4), a single-colony high-throughput growth monitoring system (8) and an automatic controller (10);
one side of the saliva sampling plate (2) is detachably connected to the oral saliva sampling device (1), and when the oral saliva sampling device (1) is placed in an oral cavity, saliva in the oral cavity is introduced into the saliva sampling plate (2) by the oral saliva sampling device (1), the automatic high-flux colony selector (3) scans colony high-flux imaging pictures on the saliva sampling plate (2) and analyzes the pictures, a selecting needle of the automatic high-flux colony selector (3) is used for completing colony selection on the saliva sampling plate (2) according to analysis results and setting colony requirements to be selected according to parameters, the selected colony samples are placed on a single colony inoculation plate (4), then the single colony inoculation plate (4) is placed in a culture dish of the single colony high-flux growth monitoring system (8), a high-flux imaging module of the single colony high-flux growth monitoring system (8) is used for scanning and monitoring growth conditions of single colonies in the culture dish in real time, monitoring results are recorded and output, and the automatic controller (10) is connected with the automatic high-flux selector (3) and the single colony high-flux monitoring system (8) respectively.
2. The high throughput sequencing-based oral flora analysis device of claim 1, wherein: the automatic high-flux colony picking device comprises a tray device (5), and is characterized by further comprising a tray device (5) and a manipulator (6), wherein the tray device (5) and the manipulator (6) are respectively arranged on one side of an automatic high-flux colony picking device (3), the tray device (5) is used for transferring saliva sampling plates (2), the manipulator (6) is used for grabbing the saliva sampling plates (2) on the tray device (5) and placing the saliva sampling plates on a colony picking station of the automatic high-flux colony picking device (3), and the automatic controller (10) controls execution actions of the manipulator (6).
3. The high throughput sequencing-based oral flora analysis device of claim 2, wherein: the tray device (5) comprises a tray frame (51) and an automatic cover plate assembly, the tray frame (51) is arranged on one side of an automatic high-flux colony selection instrument (3), a plurality of laminates (511) are arranged at intervals of the tray frame (51), the automatic cover plate assembly comprises a contact switch (52), a singlechip (53), a motor (54) and a cover plate (55), a switch groove is formed in the upper surface of the laminate (511), the contact switch (52) is arranged at the switch groove and is in control connection with the singlechip (53), a power shaft of the motor (54) is connected with one side of the cover plate (55) in a transmission mode, the singlechip (53) controls the switch of the motor (54), and when the contact switch (52) is shielded by a saliva sampling plate (2), the singlechip (53) controls the motor (54) to start, and the cover plate (55) is driven to cover the upper surface of the saliva sampling plate (2), and the automatic controller (10) is in communication connection with the singlechip (53).
4. The high throughput sequencing-based oral flora analysis device of claim 1, wherein: oral cavity saliva sampling device (1) includes facing (11), lining (12), a set of siphon connecting strip (13) and sample cardboard (14), lining (12) are laid on facing (11), and when facing (11) use, saliva is collected to lining (12) direct contact tooth, and the inboard at lining (12) is connected respectively to the one end of a set of siphon connecting strip (13), sample cardboard (14) include frame board (141) and gauze face (142), the other end of a set of siphon connecting strip (13) is connected respectively around frame board (141) to be located the intraductal intermediate position of facing (11), frame board (141) are equipped with the inside and outside passage way hole that switches on of frame, gauze face (142) set up the inboard at frame board (141), and absorb saliva that siphon connecting strip (13) guide through the passage way hole, saliva sampling board (2) are detained when dress in frame board (141), and sampling face contact gauze face (142), with saliva equipartition on the face of taking out of gauze (2).
5. The high throughput sequencing-based oral flora analysis device of claim 4, wherein: the lining cloth (12), the siphon connecting strip (13) and the gauze surface (142) are all made of medical polyester fiber cloth.
6. The high throughput sequencing-based oral flora analysis device of claim 1, wherein: the automatic high-throughput colony picking instrument is characterized in that a transfer manipulator (7) is arranged between the automatic high-throughput colony picking instrument (3) and the single-colony high-throughput growth monitoring system (8), the transfer manipulator (7) is used for transferring the single-colony inoculation plate (4) into a culture dish of the single-colony high-throughput growth monitoring system (8), and the automatic controller (10) controls the execution action of the transfer manipulator (7).
7. The high throughput sequencing-based oral flora analysis device of claim 1, wherein: the automation controller (10) is an industrial personal computer.
8. The high throughput sequencing-based oral flora analysis device of claim 1, wherein: the automatic high-throughput colony picker (3) adopts a RapidPick automatic clone colony picker.
9. An oral cavity flora analysis method based on high-throughput sequencing, which is characterized in that: the oral colony analysis using the high throughput sequencing-based oral flora analyzer of any of claims 1-8, comprising the specific steps of:
step 1, oral saliva sampling, namely placing a dental socket (11) in an oral cavity, clenching the dental socket (11) by teeth, keeping for 2-3 minutes, enabling a gauze surface (142) to be full of saliva, taking out the dental socket (11), clamping a saliva sampling plate (2) at a frame plate (141), uniformly staining saliva on the sampling surface of the saliva sampling plate (2), placing a label attached to the saliva sampling plate (2) on a layer plate (511) of a tray device (5), and starting an automatic cover plate assembly to cover the saliva sampling plate (2);
step 2, colony picking, wherein an automatic controller (10) controls a manipulator (6) to pick a designated saliva sampling plate (2), when the gripper of the manipulator (6) reaches the position of the designated saliva sampling plate (2), the automatic controller (10) controls an automatic cover plate assembly to be opened, the manipulator (6) finishes picking, the saliva sampling plate (2) is placed on a colony picking station of an automatic high-flux colony picking instrument (3), and the automatic high-flux colony picking instrument (3) respectively collects bacterial groups needing to be sampled and analyzed on the saliva sampling plate (2) into single colony inoculation plates (4) according to set parameters;
step 3, colony culture, wherein an automatic controller (10) controls a transfer manipulator (7) to transfer a single colony inoculation plate (4) into a culture dish of a single colony high-flux growth monitoring system (8) for culture;
and 4, colony detection, wherein a single-colony high-flux growth monitoring system (8) scans and monitors the growth condition of single colonies in the culture dish in real time, and outputs the recorded monitoring result to an automatic controller (10).
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311254047.6A CN116987585A (en) | 2023-09-27 | 2023-09-27 | Oral cavity flora analysis device and method based on high-throughput sequencing |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202311254047.6A CN116987585A (en) | 2023-09-27 | 2023-09-27 | Oral cavity flora analysis device and method based on high-throughput sequencing |
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| CN116987585A true CN116987585A (en) | 2023-11-03 |
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| CN202311254047.6A Withdrawn CN116987585A (en) | 2023-09-27 | 2023-09-27 | Oral cavity flora analysis device and method based on high-throughput sequencing |
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Application publication date: 20231103 |