CN116836769A - Sample processing and detection kit with liquid transfer structure - Google Patents
Sample processing and detection kit with liquid transfer structure Download PDFInfo
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- CN116836769A CN116836769A CN202210300021.XA CN202210300021A CN116836769A CN 116836769 A CN116836769 A CN 116836769A CN 202210300021 A CN202210300021 A CN 202210300021A CN 116836769 A CN116836769 A CN 116836769A
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- 239000007788 liquid Substances 0.000 title claims abstract description 100
- 238000001514 detection method Methods 0.000 title claims abstract description 54
- 238000012546 transfer Methods 0.000 title claims abstract description 42
- 238000012545 processing Methods 0.000 title claims abstract description 23
- 238000007789 sealing Methods 0.000 claims description 14
- 230000000670 limiting effect Effects 0.000 claims description 9
- 230000009471 action Effects 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 6
- 239000002861 polymer material Substances 0.000 claims description 6
- 239000000463 material Substances 0.000 claims description 5
- 230000003287 optical effect Effects 0.000 claims description 5
- 239000004743 Polypropylene Substances 0.000 claims description 4
- -1 polypropylene Polymers 0.000 claims description 4
- 229920001155 polypropylene Polymers 0.000 claims description 4
- 238000003745 diagnosis Methods 0.000 abstract description 9
- 201000010099 disease Diseases 0.000 abstract description 9
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 9
- 238000000605 extraction Methods 0.000 abstract description 6
- 238000006243 chemical reaction Methods 0.000 description 16
- 238000010438 heat treatment Methods 0.000 description 14
- 239000000243 solution Substances 0.000 description 10
- 238000004140 cleaning Methods 0.000 description 8
- 238000005516 engineering process Methods 0.000 description 7
- 102000004190 Enzymes Human genes 0.000 description 6
- 108090000790 Enzymes Proteins 0.000 description 6
- 239000003153 chemical reaction reagent Substances 0.000 description 6
- 239000002699 waste material Substances 0.000 description 6
- 230000010354 integration Effects 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
- 102100034343 Integrase Human genes 0.000 description 4
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 4
- 230000003321 amplification Effects 0.000 description 4
- 239000006166 lysate Substances 0.000 description 4
- 244000005700 microbiome Species 0.000 description 4
- 238000003199 nucleic acid amplification method Methods 0.000 description 4
- 210000002345 respiratory system Anatomy 0.000 description 4
- 238000007664 blowing Methods 0.000 description 3
- 230000005284 excitation Effects 0.000 description 3
- 150000007523 nucleic acids Chemical class 0.000 description 3
- 102000039446 nucleic acids Human genes 0.000 description 3
- 108020004707 nucleic acids Proteins 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 238000012408 PCR amplification Methods 0.000 description 2
- 230000004308 accommodation Effects 0.000 description 2
- 239000000443 aerosol Substances 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000011324 bead Substances 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 125000004122 cyclic group Chemical group 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000001917 fluorescence detection Methods 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- 238000001746 injection moulding Methods 0.000 description 2
- 229920002521 macromolecule Polymers 0.000 description 2
- 244000000010 microbial pathogen Species 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
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- 244000052769 pathogen Species 0.000 description 2
- 230000001717 pathogenic effect Effects 0.000 description 2
- 238000003753 real-time PCR Methods 0.000 description 2
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- 208000026350 Inborn Genetic disease Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 238000005299 abrasion Methods 0.000 description 1
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 1
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING 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
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/02—Burettes; Pipettes
- B01L3/021—Pipettes, i.e. with only one conduit for withdrawing and redistributing liquids
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING 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
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
Abstract
The invention provides a sample processing and detecting kit provided with a liquid transfer structure, comprising: the upper cover and the chassis form a hollow accommodating space together, and a plurality of liquid accommodating spaces are arranged on the chassis; the central rotating shaft is connected with the upper cover, the central rotating shaft is connected with the liquid transfer device, and the liquid transfer device is driven by the central rotating shaft to realize liquid transfer between different liquid accommodating spaces. All the extraction and detection steps can be carried out in the same closed space, so that the time required by detection is shortened, and the detection result is less influenced by manual operation and can be more accurate; by arranging a plurality of detection bins, the simultaneous detection of a plurality of disease types can be realized, and the diagnosis and the multiple detection of clinical similar diseases are greatly simplified.
Description
Technical Field
The invention relates to the field of biotechnology devices, in particular to a sample processing and detecting kit with a liquid transfer structure and application thereof.
Background
At present, the conventional development of diagnostic detection technology platforms mainly comprise microorganism detection, immunity detection and molecular detection. The method is mainly applied to the aspects of pathogenic microorganism infectivity detection, tumor and genetic disease diagnosis, immune system disease diagnosis, prenatal screening and the like in medical examination.
At present, the main flow technology of molecular detection is the fluorescent quantitative PCR technology, and due to the characteristics of an exponential amplification template of the PCR technology, aerosol pollution caused by open consumables and the requirement of a laboratory on liquid preparation and extraction in a partitioning way, and further high requirements on clinical environment for detection become important factors for limiting the further application of the fluorescent quantitative PCR technology, meanwhile, the traditional product has the defects of more experimental steps, complex operation, PCR aerosol pollution, low automation degree, unstable result easily caused by manual operation and high difficulty in multiple detection implementation.
With the development of technology, the demand for molecular diagnosis technology is increasing towards accuracy, convenience, sensitivity, automation and integration, but due to the complexity of molecular detection, the full automation and integration instrument platform from sample to result is less, or a plurality of technical problems such as complex structure, poor sensitivity and specificity and complex operation and detection equipment are difficult to solve.
Disclosure of Invention
Aiming at the defects that the prior art only realizes nucleic acid detection and cannot realize integration from sample processing to detection, the invention provides a sample processing and detecting kit with a liquid transfer structure and application thereof.
The invention provides a sample processing and detecting kit provided with a liquid transfer structure, comprising:
an upper cover is arranged on the upper cover,
the chassis and the upper cover form a hollow accommodating space together, and a plurality of liquid accommodating spaces are arranged on the chassis;
the central rotating shaft is connected with the upper cover, the lower part of the central rotating shaft is connected with a liquid transfer device, and the liquid transfer device is driven by the central rotating shaft to realize liquid transfer between different liquid accommodating spaces.
Wherein, the center pivot includes:
the lower end of the main rotating shaft and the liquid transferring device drive the liquid transferring device to move along the vertical direction and/or rotate around the axis under the drive of external force;
the central push rod is provided with a hollow accommodating cavity in the middle of the main rotating shaft, is positioned in the accommodating cavity and moves up and down in the accommodating cavity under the action of external force; the lower end of the central push rod is connected with a piston head, and the bottom surface of the piston head is in sealing connection with the inner wall of the accommodating cavity and the liquid transferring device;
the central rotating shaft shell comprises a cylindrical first shell and an annular second shell, the lower end of the first shell is connected with the second shell, the first shell is sleeved outside the central rotating shaft, and the second shell is connected with the liquid transferring device.
One end of the main rotating shaft is connected with the first driving device and driven by the first driving device to move in the vertical direction and/or rotate around the axis; the center push rod can be connected with the second driving device and driven by the second driving device to move along the vertical direction.
The first driving device is an outer spring, the upper end of the outer spring is fixedly connected with the main rotating shaft, and the lower end of the outer spring is fixedly connected with the first shell; the second driving device is an inner spring, the upper end of the inner spring is connected with the central push rod, and the lower end of the inner spring is connected with the inner wall of the accommodating cavity.
The upper cover is provided with an annular boss extending outwards, and the annular boss is provided with a sample inlet; the sample inlet is arranged corresponding to the liquid transferring device; and the sample inlet is provided with a sealing plug which is detachably connected with the sample inlet.
The outer ring of the chassis is provided with a clamping groove, the upper cover is provided with a buckle corresponding to the clamping groove, and the clamping groove is in detachable sealing connection with the buckle.
The chassis is provided with a first liquid containing hole, and the first liquid containing hole is close to the edge of the chassis; a second accommodating hole and a third accommodating hole are symmetrically arranged between the first accommodating hole and the central hole, a fourth accommodating hole and a fifth accommodating hole are arranged on the outer sides of the second accommodating hole and the third accommodating hole, and a plurality of seventh accommodating holes which are uniformly distributed are arranged on one side, opposite to the first accommodating hole, of the chassis; and a sixth accommodating hole is arranged in the area between the seventh accommodating hole and the fourth accommodating hole.
Wherein the chassis is integrally formed by transparent high polymer materials; the transparent polymer material is an optical polypropylene material.
And the chassis is also provided with a limiting hole, and the depth of the limiting hole is larger than that of the seventh accommodating hole.
The invention also provides a sample processing and detecting method, which uses the detecting kit and specifically comprises the following steps:
1) Sealing and storing liquid: sealing the solution needed by the reaction in the first accommodating hole, the second accommodating hole, the third accommodating hole, the fourth accommodating hole, the fifth accommodating hole and the seventh accommodating hole independently;
2) Adding a sample to be detected, wherein the sample reacts with the detection liquid;
3) And detecting the reacted solution to obtain a detection result of the sample.
Further, in the step 1, the magnetic bead solution is stored in the first containing hole; a second vessel Kong Cunfang lysate; a third container Kong Cunfang of cleaning liquid; fourth, kong Cunfang reconstitution buffer; fifth house Kong Cunfang freeze-dried reverse transcriptase; the seventh accommodation holes respectively store a plurality of freeze-dried PCR enzymes corresponding to the sample to be tested.
Further, in the step 2, the method includes:
21 A) adding a sample containing bacteria to well 1;
22 Transferring a certain amount of lysate from the well 2 to the well 1 by the suction head, and simultaneously heating the well 1 for a period of time to lyse bacteria and release RNA;
23 During the heating process of the hole 1, the suction head transfers the re-dissolving buffer solution of the hole 4 to the holes 5 and 7 to dissolve the freeze-dried enzyme therein;
24 After the heating of the hole No. 1 is finished, the suction head transfers the waste liquid of the hole No. 1 to the hole No. 2;
25 Transferring the cleaning liquid in the hole 3 to the hole 1 by the suction head, and repeatedly blowing and sucking the cleaning liquid in the hole 1 by the suction head to complete thorough cleaning;
26 Transferring the waste liquid of the hole 1 to the hole 3 by the suction head;
27 Transferring reverse transcriptase solution from well 5 to well 1 by the tip while well 1 is heated for a period of time, reverse transcribing RNA into DNA;
28 Transferring the DNA solution of the hole No. 1 to the hole No. 7, mixing with the PCR enzyme therein, and circularly heating the hole No. 7 at 50-95 ℃;
29 In the cyclic heating process of the number 7 hole, excitation light of a matched instrument is injected into the number 7 hole, fluorescence of the number 7 hole is collected, independent analysis is carried out on fluorescence signals of each hole, and finally a diagnosis result of respiratory tract pathogen microorganisms contained in a sample can be obtained.
The central rotating shaft drives the liquid-transferring suction head to rotate to the upper part of the corresponding area, and then drives the suction head to sink to puncture the aluminum film, so that the actions of sucking liquid, blowing liquid or pumping liquid to each area can be performed. Finally, the liquid flowing, mixing and reacting among the areas of different liquids are realized, and finally, the sample extracting and detecting amplification function is realized.
According to the invention, through sample treatment and detection reaction of liquid transfer in the detection box, preparation and detection of biological macromolecules such as nucleic acid and the like can be realized in a closed space as an automatic integration, and the problems of complicated extraction and detection operations, time and labor consumption, inaccurate sample or reagent pollution and the like are overcome; in addition, all the extraction and detection steps can be carried out in the same closed space, so that the time required for detection is shortened, and the detection result is less influenced by manual operation and can be more accurate; by arranging a plurality of detection bins, the simultaneous detection of a plurality of disease types can be realized, and the diagnosis and the multiple detection of clinical similar diseases are greatly simplified. The constant-temperature amplification or PCR amplification can be carried out through different detection hole settings, and the application range is wide.
Drawings
FIG. 1 is a schematic diagram of a sample processing and detection kit provided with a liquid transfer structure according to the present invention;
FIG. 2 is a schematic diagram showing the structure of a chassis of a sample processing and detecting kit provided with a liquid transfer structure according to the present invention;
FIG. 3 is a schematic view showing the structure of a chassis of the sample processing and detecting kit provided with the liquid transfer structure of the present invention.
Fig. 4 shows the detected curve in example 2 of the present invention.
Reference numerals illustrate:
10 upper housing, 11. Main shaft, 12 center pushrod, 13 piston head, 14 center shaft housing, 141 first housing, 142 second housing, 15 liquid transfer device, 16 outer spring, 17 inner spring, 18 seal ring, 9 bottom plate, 91 first receiving hole, 92 second receiving hole, 93 third receiving hole, 94 fourth receiving hole, 95 fifth receiving hole, 96 sixth receiving hole, 97 seventh receiving hole, 98 center hole, 99 limiting hole.
Detailed Description
An embodiment of the present invention will be specifically explained below with reference to the drawings.
Example 1
The invention provides a sample processing and detecting kit provided with a liquid transfer structure, comprising:
the upper cover 10 is provided with a pair of opening portions,
a chassis 9, wherein the chassis 9 and the upper cover 10 form a hollow accommodating space together, and a plurality of liquid accommodating spaces are arranged on the chassis;
the upper end of the central rotating shaft is sleeved in the through hole on the top surface of the upper cover 10, the lower part of the central rotating shaft is connected with a liquid transfer device 15, and the liquid transfer device 15 is driven by the central rotating shaft to realize liquid transfer among different liquid accommodating spaces;
the liquid transferring device 15 is a hollow shell with a funnel-shaped lower part, and a through hole is arranged at the bottom of the liquid containing device.
The center rotary shaft includes:
and one end of the main rotating shaft 11 is connected with the first driving device, and is driven by the first driving device to move in the vertical direction and/or rotate around the axis. The first driving device can be a driving motor, and the driving motor drives the main rotating shaft to move up and down or rotate around the axis. The first driving device can be an outer spring, the upper end of the outer spring is fixedly connected with the main rotating shaft, the lower end of the outer spring is fixedly connected with the upper cover, the main rotating shaft moves downwards under the action of external force, the outer spring is compressed, and after the external force is removed, the outer spring is restored and pushes the main rotating shaft to move upwards to restore to the initial position.
The main rotating shaft 11 is provided with a circle of annular bulge at a position matched with the inner wall of the upper cover 10, and a sealing ring is arranged between the annular bulge and the upper cover. The annular bulge and the sealing ring are designed, so that the limiting effect of the upper cover on the upper end of the main rotating shaft can be guaranteed, and abrasion caused by direct contact of an interaction part between the upper cover and the main rotating shaft can be avoided.
The center push rod 12, the middle part of main pivot 11 has hollow accommodation chamber, center push rod 12 is located hold the intracavity, center push rod 12 is under the exogenic action in hold the intracavity and reciprocate, the lower extreme of center push rod 12 is connected with piston head 13, the bottom surface of piston head 13 holds and hold between intracavity wall and the liquid transfer device 15 sealing connection.
The center pushrod 12 may be connected to a second driving device, which may drive a motor, an air pump, or a hydraulic pump. The second driving device is an inner spring, the upper end of the inner spring is connected with the central push rod 12, the lower end of the inner spring is connected with the inner wall of the accommodating cavity, under the action of external force, the central push rod 12 moves downwards and enables the inner spring to be compressed, at the moment, the piston moves downwards, air in the liquid transferring device is discharged outwards, after the external force is removed, the inner spring is restored and pushes the central push rod to move upwards to restore to the initial position, at the moment, the piston moves upwards, negative pressure is formed in the space between the bottom surface of the piston head, the inner wall of the accommodating cavity and the liquid transferring device, and liquid is sucked into the liquid transferring device.
The center of the chassis is provided with a through hole, and the center rotating shaft penetrates through the through hole. The central shaft portion is sunk into the chassis so that the overall height of the kit can be reduced, but is not connected to the chassis.
The rotary shaft type liquid transfer device comprises a rotary shaft, and is characterized by further comprising a central rotating shaft shell, wherein the rotary shaft shell comprises a cylindrical first shell and an annular second shell, the upper end of the first shell is connected with an upper cover, the lower end of the first shell is connected with the second shell, the first shell is sleeved outside the central rotating shaft, and the second shell is connected with the liquid transfer device
The outer edge of the second shell is provided with a clamping groove, the liquid transferring device is detachably connected in the clamping groove, and a sealing gasket is arranged in the clamping groove.
An annular boss extending outwards is arranged on the upper cover, and a sample inlet is arranged on the annular boss; the sample inlet is arranged corresponding to the liquid transferring device.
The outer lane of chassis is equipped with the draw-in groove, be equipped with on the upper shield with the draw-in groove corresponding buckle, draw-in groove and buckle detachable sealing connection.
The chassis is provided with a first liquid accommodating hole 91, and the first liquid accommodating hole 91 is close to the edge of the chassis; a second accommodating hole 92 and a third accommodating hole 93 are symmetrically arranged between the first accommodating hole 91 and the central hole 98, a fourth accommodating hole 94 and a fifth accommodating hole 95 are arranged on the outer sides of the second accommodating hole 92 and the third accommodating hole 93, and a plurality of 7 th accommodating holes 97 which are uniformly distributed are arranged on the opposite side of the chassis to the first accommodating hole 91; a sixth receiving hole 96 is provided in a region between the seventh receiving hole 97 and the fourth receiving hole 94.
The horizontal distance between the through hole and the axis of the central rotating shaft is L1, the minimum horizontal distance between each containing hole on the chassis and the center of the chassis is Lnmin, the maximum horizontal distance between each containing hole on the chassis and the center of the chassis is Lnmax, lnmin < L1 < Lnmax, wherein n=1, 2,3,4,5,6,7.
The chassis 9 is integrally formed by transparent high polymer materials. The transparent polymer material can be an optical polypropylene material. The chassis 9 is formed by injection molding of an optical polypropylene material, has high transparency and stable chemical characteristics, is mainly used for storage and reaction of various reagent solutions, and can be used for fluorescence detection. The chassis 9 is provided with a liquid storage cavity and a reaction cavity in various shapes, the liquid storage cavity is mainly used for storing reagent solution and waste liquid in the reaction process, and the reaction cavity is mainly used for amplifying and reacting various reagents. The liquid storage cavity is large in general volume, the reaction cavity is conical in outline shape, similar to a PCR tube, and the liquid storage cavity is beneficial to being tightly attached to the heating block, so that the heating efficiency is improved.
The outer wall of each storage area on the chassis is thicker and is used for preventing liquid from permeating or volatilizing; the outer wall of the reaction zone is designed to be thin in order to increase the heat transfer efficiency when heating. The chassis is integrally formed by injection molding of high-transparency high polymer materials and has good optical permeability, so that the device can be used for excitation and reception of reaction fluorescence.
The second, third, fourth and fifth receiving holes may be a liquid storage area or a waste liquid area. Each containing hole part is designed with a film sealing ring structure, and if the area is set as a storage area, the film sealing can be performed.
The first accommodating hole, the sixth accommodating hole and the seventh accommodating hole are liquid storage areas or reaction areas. It should be noted here that the storage area and the reaction area are not strictly defined between the liquid containing holes, and any area may be used as the storage area or the reaction area.
The reaction zone can be provided with heating equipment in a matched manner.
The central hole 98 is mainly used for fixing the lower end of the central rotating shaft, so that the central rotating shaft is beneficial to sinking and sucking liquid, and the whole height of the kit can be reduced. The limiting hole 99 is designed to raise the seventh receiving hole 97 (10-joint hole), so that it is necessary to preserve a high surface finish in view of the fact that the seventh receiving hole 97 is mainly used for fluorescence detection, and the seventh receiving hole 97 is raised by lengthening the limiting hole 99, which can prevent the seventh receiving hole 97 from rubbing against the table top, thereby generating scratches.
The application range of the sample processing and detection integrated kit provided with the liquid transfer structure includes but is not limited to: the method is suitable for the integrated design of microorganism, immunity and molecular detection kits.
Taking simultaneous detection of various diseases of respiratory tract as an example, the kit is used, and the specific method comprises the following steps:
1) And (3) liquid packaging and storing: the first accommodating hole is used for storing the magnetic bead solution; a second vessel Kong Cunfang lysate; a third container Kong Cunfang of cleaning liquid; fourth, kong Cunfang reconstitution buffer; fifth house Kong Cunfang freeze-dried reverse transcriptase; the seventh receiving well (10 wells) stores respective lyophilized PCR enzymes for a plurality of pathogenic microorganisms of the respiratory tract.
22 Reaction process:
21 A) adding a sample containing bacteria to well 1;
22 Transferring a certain amount of lysate from the well 2 to the well 1 by the suction head, and simultaneously heating the well 1 for a period of time to lyse bacteria and release RNA;
23 During the heating process of the hole 1, the suction head transfers the re-dissolving buffer solution of the hole 4 to the holes 5 and 7 to dissolve the freeze-dried enzyme therein;
24 After the heating of the hole No. 1 is finished, the suction head transfers the waste liquid of the hole No. 1 to the hole No. 2;
25 Transferring the cleaning liquid in the hole 3 to the hole 1 by the suction head, and repeatedly blowing and sucking the cleaning liquid in the hole 1 by the suction head to complete thorough cleaning;
26 Transferring the waste liquid of the hole 1 to the hole 3 by the suction head;
27 Transferring reverse transcriptase solution from well 5 to well 1 by the tip while well 1 is heated for a period of time, reverse transcribing RNA into DNA;
28 Transferring the DNA solution of the hole No. 1 to the hole No. 7, mixing with the PCR enzyme therein, and circularly heating the hole No. 7 at 50-95 ℃;
29 In the cyclic heating process of the number 7 hole, excitation light of a matched instrument is injected into the number 7 hole, fluorescence of the number 7 hole is collected, independent analysis is carried out on fluorescence signals of each hole, and finally a diagnosis result of respiratory tract pathogen microorganisms contained in a sample can be obtained.
According to the invention, through sample treatment and detection reaction of liquid transfer in the detection box, preparation and detection of biological macromolecules such as nucleic acid and the like can be realized in a closed space as an automatic integration, and the problems of complicated extraction and detection operations, time and labor consumption, inaccurate sample or reagent pollution and the like are overcome; in addition, all the extraction and detection steps can be carried out in the same closed space, so that the time required for detection is shortened, and the detection result is less influenced by manual operation and can be more accurate; by arranging a plurality of detection bins, the simultaneous detection of a plurality of disease types can be realized, and the diagnosis and the multiple detection of clinical similar diseases are greatly simplified. The constant-temperature amplification or PCR amplification can be carried out through different detection hole settings, and the application range is wide.
Example 2
An appropriate amount of reaction reagent (shown in table 1) is put into each hole in the chassis, after 500ul of positive sample is added, the device is put into corresponding equipment, the detected curve is shown in fig. 4, and as can be seen from fig. 4, the invention can realize simultaneous detection of a plurality of disease types by arranging a plurality of detection bins, and greatly simplifies diagnosis and multiple detection of clinical similar diseases.
TABLE 1
The above description of the specific embodiments of the present invention has been given by way of example only, and the present invention is not limited to the above-described specific embodiments. Any equivalent modifications and substitutions for the present invention will occur to those skilled in the art, and are intended to be within the scope of the present invention. Accordingly, equivalent changes and modifications are intended to be included within the scope of the present invention without departing from the spirit and scope thereof. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (10)
1. A sample processing and testing kit provided with a liquid transfer structure, comprising:
an upper cover is arranged on the upper cover,
the chassis and the upper cover form a hollow accommodating space together, and a plurality of liquid accommodating spaces are arranged on the chassis;
the central rotating shaft is connected with the upper cover, the lower part of the central rotating shaft is connected with a liquid transfer device, and the liquid transfer device is driven by the central rotating shaft to realize liquid transfer between different liquid accommodating spaces.
2. A sample processing and testing kit provided with a liquid transfer structure according to claim 1, wherein the central spindle comprises:
the lower end of the main rotating shaft and the liquid transferring device drive the liquid transferring device to move along the vertical direction and/or rotate around the axis under the drive of external force;
the central push rod is provided with a hollow accommodating cavity in the middle of the main rotating shaft, is positioned in the accommodating cavity and moves up and down in the accommodating cavity under the action of external force; the lower end of the central push rod is connected with a piston head, and the bottom surface of the piston head is in sealing connection with the inner wall of the accommodating cavity and the liquid transferring device;
the central rotating shaft shell comprises a cylindrical first shell and an annular second shell, the lower end of the first shell is connected with the second shell, the first shell is sleeved outside the central rotating shaft, and the second shell is connected with the liquid transferring device.
3. A sample processing and testing kit provided with a liquid transfer structure according to claim 2, wherein one end of the main shaft is connected to the first driving means and is driven by the first driving means to move in a vertical direction and/or rotate about an axis; the center push rod can be connected with the second driving device and driven by the second driving device to move along the vertical direction.
4. The sample processing and detecting kit with a liquid transfer structure according to claim 3, wherein the first driving device is an outer spring, the upper end of the outer spring is fixedly connected with the main rotating shaft, and the lower end of the outer spring is fixedly connected with the first housing; the second driving device is an inner spring, the upper end of the inner spring is connected with the central push rod, and the lower end of the inner spring is connected with the inner wall of the accommodating cavity.
5. The sample processing and detection kit with the liquid transfer structure according to claim 1, wherein the upper cover is provided with an annular boss extending outwards, and the annular boss is provided with a sample inlet; the sample inlet is arranged corresponding to the liquid transferring device; and the sample inlet is provided with a sealing plug which is detachably connected with the sample inlet.
6. The sample processing and detecting kit with the liquid transferring structure according to claim 1, wherein the outer ring of the chassis is provided with a clamping groove, the upper cover is provided with a buckle corresponding to the clamping groove, and the clamping groove is detachably and hermetically connected with the buckle.
7. The sample processing and testing kit with a liquid transfer structure of claim 1, wherein the chassis is provided with a first liquid receiving aperture, the first liquid receiving aperture being proximate an edge of the chassis; a second accommodating hole and a third accommodating hole are symmetrically arranged between the first accommodating hole and the central hole, a fourth accommodating hole and a fifth accommodating hole are arranged on the outer sides of the second accommodating hole and the third accommodating hole, and a plurality of seventh accommodating holes which are uniformly distributed are arranged on one side, opposite to the first accommodating hole, of the chassis; and a sixth accommodating hole is arranged in the area between the seventh accommodating hole and the fourth accommodating hole.
8. The sample processing and testing kit with a liquid transfer structure according to claim 7, wherein the chassis is integrally formed of a transparent polymeric material; the transparent polymer material is an optical polypropylene material.
9. The sample processing and testing kit with a liquid transfer structure according to any one of claims 5 to 8, wherein the chassis is further provided with a limiting hole, and the depth of the limiting hole is greater than the depth of the seventh receiving hole.
10. A method for sample processing and detection, characterized in that the detection kit according to any one of claims 1-9 is used.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210300021.XA CN116836769A (en) | 2022-03-25 | 2022-03-25 | Sample processing and detection kit with liquid transfer structure |
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| CN202210300021.XA CN116836769A (en) | 2022-03-25 | 2022-03-25 | Sample processing and detection kit with liquid transfer structure |
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| CN116836769A true CN116836769A (en) | 2023-10-03 |
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| CN202210300021.XA Pending CN116836769A (en) | 2022-03-25 | 2022-03-25 | Sample processing and detection kit with liquid transfer structure |
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