CN111534423A - Group nucleic acid testing and mixing device and method - Google Patents

Group nucleic acid testing and mixing device and method Download PDF

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Publication number
CN111534423A
CN111534423A CN202010399004.7A CN202010399004A CN111534423A CN 111534423 A CN111534423 A CN 111534423A CN 202010399004 A CN202010399004 A CN 202010399004A CN 111534423 A CN111534423 A CN 111534423A
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bottle
sampling
mixing
positioning
nucleic acid
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李文胜
伍卫国
刘松
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Xian Jiaotong University
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Xian Jiaotong University
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    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/02Burettes; Pipettes
    • B01L3/0289Apparatus for withdrawing or distributing predetermined quantities of fluid
    • B01L3/0293Apparatus for withdrawing or distributing predetermined quantities of fluid for liquids
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6806Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay

Abstract

The invention discloses a group nucleic acid testing and mixing device and a method, which comprises a sampling bottle positioning and placing disc, a mixing bottle placing disc, a sterilizing device, an automatic sucking device and a rotary control table for positioning and rotating the automatic sucking device, wherein the sampling bottle positioning and placing disc, the mixing bottle placing disc and the sterilizing device are fixed on one side of the rotary control table, a plurality of sampling bottle placing grooves are arranged on the sampling bottle positioning and placing disc, and a mechanical arm which can drive the automatic sucking device to rotate and move on the rotary control table is utilized to realize the rapid positioning and sucking of a sample to be detected stored in a sampling bottle to be detected by the automatic sucking device, so that a plurality of detection samples are mixed, the detection efficiency is improved by detecting mixed sample liquid, the device has simple structure and high safety, the sample is prevented from being mistakenly placed and taken due to the error of workers, and the detection safety of the detection samples is improved, meanwhile, the manual contact times are avoided in the mixing process, and the use safety is improved.

Description

Group nucleic acid testing and mixing device and method
Technical Field
The invention belongs to a medical detection device, and particularly relates to a group nucleic acid testing and mixing device and method.
Background
COVID-19 has rolled over the world, and the WHO suggests using a evidence-based model to deal with epidemic situations, emphasizes 'testing', and realizes virus detection confirmation through testing. However, because the virus infectivity is very strong, theoretically, synchronous testing is needed, namely, the confirmation of a large number of samples is completed within a short time, namely, the confirmation efficiency of the virus samples is improved, the virus is prevented from being rapidly diffused, and the infection of tested groups needs to be detected again, so that the confirmation efficiency of virus detection is reduced.
Disclosure of Invention
The invention aims to provide a colony nucleic acid testing mixing device and a colony nucleic acid testing mixing method, so as to overcome the defects of the prior art.
In order to achieve the purpose, the invention adopts the following technical scheme:
a group nucleic acid testing and mixing device comprises a sampling bottle positioning and placing disc, a mixing bottle placing disc, a sterilizing device, an automatic sucking device and a rotary control table for positioning and rotating the automatic sucking device, wherein the sampling bottle positioning and placing disc, the mixing bottle placing disc and the sterilizing device are fixed on one side of the rotary control table; the rotary control table is provided with a mechanical arm capable of driving the automatic suction device to rotate and move, and the sampling bottle positioning and placing disc, the mixing bottle placing disc and the sterilizing device are fixed in the automatic suction device to move, take and place the liquid within the range.
Further, sampling bottle standing groove array is on sampling bottle location place dish, and every sampling bottle standing groove has relative positioning position number with rotary control platform.
Furthermore, a positioning and placing sleeve is arranged in the sampling bottle placing groove.
Further, the positioning placing sleeve is a rubber sleeve, and the positioning placing sleeve is connected with the wall of the sampling bottle placing groove in an adhesive mode.
Further, degassing unit includes clean water basin, effluent water sump and disinfection pond, and automatic suction means can pass through the arm and drive entering clean water basin, effluent water sump and disinfection pond.
Furthermore, the automatic suction device comprises a needle tube, a needle tube clamp and an electric push rod, the needle tube clamp and the electric push rod are fixed at the end part of the mechanical arm through a fixing plate, the end part of a piston rod of the needle tube is fixed on the needle tube clamp, the clamp is fixedly clamped on the outer wall of the needle tube, and the clamp is fixed at the end part of the electric push rod through a clamping rod.
Furthermore, an opening is formed in one side of the needle tube clamp, a clamping groove is formed in the bottom of the side, provided with the opening, of the needle tube clamp, the end portion of the piston rod is clamped in the clamping groove, and the end portion of the clamping groove is locked through a screw.
Further, a mechanical arm on the rotating control table is positioned by coordinate positioning or image recognition.
A method for testing a population of nucleic acids comprising the steps of:
step 1), a plurality of sampling bottles to be detected with samples to be detected are numbered and are sequentially placed in sampling bottle placing grooves of a sampling bottle positioning placing disc;
step 2), extracting a sampling liquid from each sampling bottle to be detected in sequence by using an automatic suction device, then putting the sampling liquid into a mixing bottle to obtain a mixed sampling liquid, and disinfecting the automatic suction device once after the sampling liquid in each sampling bottle to be detected is put into the mixing bottle;
and 3), detecting the mixed sampling liquid in the mixed bottle, if the test result is negative, finishing the detection, if the test result is positive, dividing the plurality of sampling bottles to be detected 6 collected at this time into two half areas according to a halving dyeing method, and repeating the steps 1) to 3) for each half area to detect until the minimum half area unit is one sampling bottle to be detected, thus finishing the detection of a plurality of detection samples.
Further, PCR is adopted to carry out amplification test on the sampling liquid in the mixing bottle.
Compared with the prior art, the invention has the following beneficial technical effects:
the invention relates to a group nucleic acid testing and mixing device, which comprises a sampling bottle positioning and placing disc, a mixing bottle placing disc, a sterilizing device, an automatic suction device and a rotary control table for positioning and rotating the automatic suction device, wherein the sampling bottle positioning and placing disc, the mixing bottle placing disc and the sterilizing device are fixed on one side of the rotary control table, a plurality of sampling bottle placing grooves are arranged on the sampling bottle positioning and placing disc, and a mechanical arm which can drive the automatic suction device to rotate and move on the rotary control table is utilized to realize the rapid positioning and suction of a sample to be detected stored in a sampling bottle to be detected by the automatic suction device, so that a plurality of detection samples are mixed, the detection efficiency is improved by detecting a mixed sample liquid, the device has simple structure and high safety, the phenomenon that the sample is mistakenly placed and mistakenly taken due to the error of a worker is avoided, the detection safety of the detection samples is improved, and meanwhile, the manual, the use safety is improved.
Further, the cover is placed in the location and adopts the rubber sleeve, and the cover is placed in the location and sampling bottle standing groove cell wall adhesive bonding has ensured to detect the stability of placing of sampling bottle.
Furthermore, the needle tube is sucked by the needle tube clamp and the electric push rod, the structure is simple, the common needle tube can be matched for use, the application range is wide, and the emergency use efficiency of the device is improved.
Furthermore, a group nucleic acid testing method comprises the steps of sequentially placing a plurality of sampling bottles to be tested containing samples to be tested into a sampling bottle placing groove of a sampling bottle positioning placing disc, sequentially extracting sampling liquid from each sampling bottle to be tested by using an automatic suction device, placing the sampling liquid into a mixing bottle to obtain mixed sampling liquid, detecting the mixed sampling liquid in the mixing bottle, and for the mixed sampling with a positive test result, dividing the plurality of sampling bottles to be tested collected into two half areas by adopting a middle division dyeing method, sequentially detecting until the minimum half area unit is one sampling bottle to be tested, rapidly inspecting the detection of a plurality of detection samples by adopting the method, rapidly inspecting all communities with zero infection and few infection, and increasing the testing speed of the detection samples with few infection by using geometric progression, the detection efficiency is improved.
Drawings
Fig. 1 is a schematic structural diagram in an embodiment of the present invention.
FIG. 2 is a top view of a structure in an embodiment of the invention.
Fig. 3 is a schematic structural diagram of an automatic suction device in an embodiment of the present invention.
FIG. 4 is a diagram illustrating a test structure of a fractionated dye group according to an embodiment of the present invention.
In the figure, 1, a sampling bottle positioning and placing disc; 2. an automatic suction device; 3. rotating the console; 4. a mechanical arm; 5. a sampling bottle placing groove; 6. a sampling bottle to be detected; 7. a mixing bottle placing tray; 8. a sterilizing device; 9. mixing the bottles; 10. a clean water tank; 11. a sewage tank; 12. disinfecting the water tank; 13. a camera; 14. a light source; 15. a needle tube clamp; 16. an electric push rod; 17. a needle tube; 18. a screw; 19. a clamp; 20. a fixing plate; 21. a clamping rod; 22. a piston rod; 23. a support rod.
Detailed Description
The invention is described in further detail below with reference to the accompanying drawings:
as shown in fig. 1 and 2, a group nucleic acid testing and mixing device comprises a sampling bottle positioning and placing disc 1, a mixing bottle placing disc 7, a sterilizing device 8, an automatic sucking device 2 and a rotating control table 3 for positioning and rotating the automatic sucking device 2, wherein the sampling bottle positioning and placing disc 1, the mixing bottle placing disc 7 and the sterilizing device 8 are fixed on one side of the rotating control table 3, a plurality of sampling bottle placing grooves 5 are formed in the sampling bottle positioning and placing disc 1, sampling bottles 6 to be detected are placed in the sampling bottle placing grooves 5, and samples to be detected are stored in the sampling bottles 6 to be detected; be equipped with on the rotary control platform 3 and drive the arm 4 of 2 rotary motion of automatic suction means, dish 1 is placed in the sampling bottle location, mix the bottle and place dish 7 and degassing unit 8 and remove at automatic suction means 2 and get and put the liquid within range, arm 4 can drive automatic suction means 2 and absorb sampling bottle location and place and wait to detect the liquid in the sampling bottle 6 on the dish 1, and in placing mixing bottle 9 on the dish 7 with liquid removal, degassing unit 8 is used for the disinfection of automatic suction means 2. Be equipped with the rubber cap on waiting to detect sampling bottle 6, after the long needle of cylinder is pierced and is taken out, self-sealing thorn hole prevents that the sampling liquid from volatilizing.
Sampling bottle standing groove 5 array is placed on dish 1 in the sampling bottle location, and every sampling bottle standing groove 5 has the location position serial number, is equipped with the location in the sampling bottle standing groove 5 and places the cover for wait to detect sampling bottle 6 location and place, prevent to detect sampling bottle 6 and get the liquid in-process and rock, the location is placed the cover and is adopted the rubber sleeve, and the location is placed cover and 5 cell wall adhesive connections of sampling bottle standing groove.
The sterilizing device 8 comprises a clean water tank 10, a sewage tank 11 and a sterilizing water tank 12, and the automatic suction device 2 can be driven by the mechanical arm 4 to enter the clean water tank 10, the sewage tank 11 and the sterilizing water tank 12 for sterilizing the automatic suction device 2. The clean water tank 10, the sewage tank 11 and the disinfection water tank 12 can be independently integrated into a whole, so that uniform disinfection treatment can be conveniently carried out after detection, and the use safety of equipment is improved; the clean water tank 10, the sewage tank 11 and the disinfection water tank 12 can also be integrated with the sampling bottle positioning and placing disc 1, so that the uniform disinfection treatment of the equipment is facilitated after a batch of samples are detected.
As shown in fig. 3, the automatic suction device 2 includes a needle tube 17, a needle tube clamp 15 and an electric push rod 16, the needle tube clamp 15 and the electric push rod 16 are fixed at the end of the mechanical arm 4 through a fixing plate 20, the end of a piston rod 22 of the needle tube 17 is fixed on the needle tube clamp 15, a clamp 19 is fixedly clamped on the outer wall of the needle tube 17, the clamp 19 is fixed at the end of the electric push rod 16 through a clamping rod 21, and the electric push rod 16 drives the needle tube of the needle tube 17 to move through the clamp 19, so that the needle head at the end of the needle tube 17 sucks the liquid in the sample bottle. In the using process, the mechanical arm 4 drives the needle cylinder of the needle tube 17 to enable the needle head to penetrate into the sampling bottle 6 to be detected, then the mechanical arm 4 is lifted, and the electric push rod 16 pushes the needle cylinder of the needle tube 17 to move, so that the needle cylinder of the needle tube 17 and the piston rod 22 generate relative movement, and liquid is sucked.
An opening is formed in one side of the needle tube clamp 15, a clamping groove is formed in the bottom of the side, provided with the opening, of the needle tube clamp 15, the end portion of the piston rod is clamped in the clamping groove, and the end portion of the clamping groove is locked and positioned through a screw rod 18.
A mechanical arm 4 on the rotary control table 3 is positioned by coordinate positioning or image recognition; adopt coordinate location, use the rotation center of rotating control platform 3 as the coordinate center promptly, sampling bottle locating slot 5 on the dish 1 is placed in the sampling bottle location, the mixing bottle on the dish 7 is placed to the mixing bottle and 8 coordinate position fixes of degassing unit. The method comprises the steps of adopting image recognition positioning, comprising a camera 13 and a light source 14, arranging a rubber bottle cap on a sampling bottle 6 to be detected, wherein the color of the rubber bottle cap is different from the colors of other positions, and the rubber bottle cap is used for the positioning recognition of the camera 13, carrying out visual recognition according to an image shot by the camera, positioning the sampling bottle 6 to be detected, moving an automatic suction device 2 to the position above the sampling bottle 6 to be detected by a mechanical arm 4, carrying out the position and posture adjustment of a robot hand according to the shot image, enabling a needle cylinder of the automatic suction device 2 to be arranged right above the sampling bottle 6 to be detected, then pressing down, inserting a needle head into the sampling bottle 6 to be detected, controlling an electric push rod of the automatic suction device 2, moving a needle cylinder piston of; the volume of the sampling liquid meets the requirement of detecting the minimum liquid amount every time, namely, the liquid sucked from a single sampling bottle 6 to be detected can represent a detection sample in the sampling bottle 6 to be detected every time, so that the sample identification is realized; the volume of mixing bottle 9 is greater than all volume of waiting to detect sampling bottle 6 sampling liquid and, can realize that liquid mixing detects and accomplishes the detection that waits to detect sampling bottle 6 on whole sampling bottle location place dish 1. And (3) setting the liquid volume of each sampling bottle 6 to be detected as N, sucking the liquid volume from the sampling bottle 6 to be detected as 1/N each time, and injecting the sampling liquid into the mixing bottle. The sample sampling of a sampling bottle 6 to be detected is completed, the automatic suction device 2 is disinfected once, a needle cylinder of the automatic suction device 2 is moved to a clean water tank 10, clean water is sucked, the needle cylinder is moved to a sewage tank 11 to be extruded, the sewage tank is repeatedly extruded for a plurality of times, then the needle cylinder is moved to a disinfection water tank 12 to be sucked, sterile water is moved to a sewage tank to be extruded, the needle cylinder is repeatedly extruded for a plurality of times, the needle cylinder is finally moved to the clean water tank 10, clean water is sucked, the needle cylinder is rotated to the sewage tank to be extruded, the disinfectant is repeatedly extruded for.
A light source 14 is arranged on one side of the camera 13 and used for camera recognition and light supplement; the camera 13 and the light source 14 are fixed to the rotating console 3 through a support rod 23.
A population nucleic acid testing method based on the population nucleic acid testing mixing device comprises the following steps:
step 1), a plurality of sampling bottles 6 to be detected with samples to be detected are numbered and are sequentially placed in a sampling bottle placing groove 5 of a sampling bottle positioning placing disc 1;
step 2), extracting a sampling liquid from each sampling bottle 6 to be detected in sequence by using the automatic suction device 2, then putting the sampling liquid into the mixing bottle 9 to obtain a mixed sampling liquid, and sterilizing the automatic suction device 2 once after the sampling liquid in each sampling bottle 6 to be detected is put into the mixing bottle 9;
and 3), detecting the mixed sampling liquid in the mixed bottle 9, if the test result is negative, finishing the detection, if the test result is positive, dividing the plurality of sampling bottles to be detected 6 collected at this time into two half areas according to a halving dyeing method, and repeating the steps 1) to 3) for each half area to detect until the minimum half area unit is one sampling bottle to be detected 6, namely finishing the detection of a plurality of detection samples. Such as: if the first detection result of the sampling liquid in the mixing bottle 9 is positive, dividing the plurality of sampling bottles 6 to be detected collected into an upper half area and a lower half area according to a middle division dyeing method, respectively detecting the upper half area and the lower half area, and if the upper half area is positive and the lower half area is negative, excluding the possibility that the sample in the lower half area is positive; the upper half area was tested by the median staining method.
This application adopts PCR to carry out the amplification test to the sampling liquid in the mixing bottle 9, compares with monomer test, needs increase K amplification, if the colony quantity in the mixing bottle 9 is m, then K is log ═ log2m, so that the sensitivity is the same as that of the monomer test, and the value of the maximum population number m is adjusted according to the actually-achieved sensitivity, so that the test accuracy reaches the requirement.
As shown in fig. 4, the method of the medium division dyeing specifically includes the following steps:
TC communities or groups are arranged, the testing times of each community are CS, and the total testing times are as follows:
Figure BDA0002488753740000071
wherein the test total CS [ i ] of the ith community is:
Figure BDA0002488753740000081
m [ i ] in formula 2]The number of people in the i community is the total number of samples, if 2k-1<m[i]<2kThen take m [ i ]]=2k. j represents the jth test, after each test round is finished, the areas without problems are dyed with ink marks, and the areas are not tested in the next test round. The plot is for the case where m is 64, assuming 5 infected samples, white in fig. 4, and the other areas are stained black one by one during the test.
As can be seen from FIG. 4, in the best case, 0 cases of infection in the community need to be measured only 1 time, and if there are 1 case, 2 logs are needed2m +1 times, while the traditional monomer test requires m times, in the case of m 1024, the split dyeing method requires 21 times, and the monomer test requires 1024 times. Therefore, the method can be used for rapidly checking communities with less infection number, is particularly suitable for countries and regions with less infection number at the beginning of infection, can be used for rapidly checking all communities with zero infection and less infection number, and can be used for performing monomer test on a few communities with more infection number. The invention improves the testing speed of the population with less infection quantity by geometric progression, thereby being capable of rapidly carrying out global investigation. Compared with the development of vaccines and specific drugs, the visual recognition automatic control and amplification test are mature technologies, the development time and accuracy are controllable, and the control can be fast realized when the epidemic situation is initiated.

Claims (10)

1. The group nucleic acid testing and mixing device is characterized by comprising a sampling bottle positioning and placing disc (1), a mixing bottle placing disc (7), a sterilizing device (8), an automatic suction device (2) and a rotary control table (3) for positioning and rotating the automatic suction device (2), wherein the sampling bottle positioning and placing disc (1), the mixing bottle placing disc (7) and the sterilizing device (8) are fixed on one side of the rotary control table (3), a plurality of sampling bottle placing grooves (5) are formed in the sampling bottle positioning and placing disc (1), sampling bottles (6) to be detected are placed in the sampling bottle placing grooves (5), and samples to be detected are stored in the sampling bottles (6) to be detected; be equipped with on rotary control platform (3) and can drive arm (4) of automatic suction means (2) swivelling movement, sampling bottle location is placed dish (1), is mixed bottle and is placed dish (7) and degassing unit (8) and be fixed in automatic suction means (2) and remove and get in the liquid scope.
2. A colony nucleic acid test mixing apparatus according to claim 1, wherein the sample bottle placement slots (5) are arrayed on the sample bottle positioning placement tray (1), and each sample bottle placement slot (5) has a relative positioning position number with the rotary console (3).
3. The colony nucleic acid test mixing arrangement of claim 1, wherein, the sample bottle standing groove (5) is provided with a positioning and placing sleeve.
4. The colony nucleic acid test mixing arrangement of claim 3, characterized in that, the location is placed the cover and is adopted the rubber sleeve, and the location is placed the cover and is placed the groove wall adhesive joint of groove (5) with the sample bottle.
5. The colony nucleic acid testing mixing device according to claim 1, characterized in that the disinfection device (8) comprises a clean water basin (10), a sewage basin (11) and a disinfection water basin (12), and the automatic suction device (2) can be driven by the mechanical arm (4) to enter the clean water basin (10), the sewage basin (11) and the disinfection water basin (12).
6. The group nucleic acid testing and mixing device of claim 1, wherein the automatic sucking device (2) comprises a needle tube (17), a needle tube clamp (15) and an electric push rod (16), the needle tube clamp (15) and the electric push rod (16) are fixed at the end of the mechanical arm (4) through a fixing plate (20), the end of a piston rod (22) of the needle tube (17) is fixed on the needle tube clamp (15), a clamp (19) is fixedly clamped on the outer wall of the needle tube (17), and the clamp (19) is fixed at the end of the electric push rod (16) through a clamping rod (21).
7. The device for testing and mixing colony nucleic acid as claimed in claim 1, wherein the needle tube fixture (15) is provided with an opening at one side, the needle tube fixture (15) is provided with a slot at the bottom of the opening, the end of the piston rod is clamped in the slot, and the end of the slot is locked by the screw (18).
8. The apparatus for mixing population nucleic acid tests according to claim 1, wherein the robotic arm (4) on the rotating console (3) is positioned using coordinate positioning or image recognition.
9. A method for testing a population nucleic acid based on the device for mixing a population nucleic acid test according to claim 1, comprising the steps of:
step 1), a plurality of sampling bottles to be detected with samples to be detected are numbered and are sequentially placed in sampling bottle placing grooves of a sampling bottle positioning placing disc;
step 2), extracting a sampling liquid from each sampling bottle to be detected in sequence by using an automatic suction device, then putting the sampling liquid into a mixing bottle to obtain a mixed sampling liquid, and disinfecting the automatic suction device once after the sampling liquid in each sampling bottle to be detected is put into the mixing bottle;
and 3), detecting the mixed sampling liquid in the mixed bottle, if the test result is negative, finishing the detection, if the test result is positive, dividing the plurality of sampling bottles to be detected 6 collected at this time into two half areas according to a halving dyeing method, and repeating the steps 1) to 3) for each half area to detect until the minimum half area unit is one sampling bottle to be detected, thus finishing the detection of a plurality of detection samples.
10. The method for testing population nucleic acid of claim 9, wherein the sample fluid in the mixing vial is subjected to an amplification test using PCR.
CN202010399004.7A 2020-05-12 2020-05-12 Group nucleic acid testing and mixing device and method Pending CN111534423A (en)

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