CN113355393A - Nucleic acid detection method based on metal coding technology - Google Patents
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Abstract
The invention provides a nucleic acid detection method based on a metal coding technology, which specifically comprises the following steps: encoding magnetic beads by using metal, and coupling a probe of a target gene with the magnetic beads to obtain metal-encoded probe magnetic beads; extracting genes, and amplifying by using a primer modified with biotin; co-incubating the metal-encoded probe magnetic beads and the amplified gene; adding metal-labeled avidin to combine with the biotin, removing redundant components, and then sending to a mass spectrometer for detection; the metal labeling the avidin is different from the metal encoding the magnetic beads; obtaining the content of metal ions in the avidin by using a mass spectrometry technology; and obtaining the information of the gene by using the obtained metal ion content and the mapping relation, wherein the mapping relation is the corresponding relation between the metal ion content and the information of the gene. The invention has the advantages of high detection accuracy, rapidness and the like.
Description
Technical Field
The invention relates to nucleic acid detection, in particular to a nucleic acid detection method based on a metal coding technology.
Background
With the completion of the Human Genome Project (HGP) and the availability of a vast amount of genetic information, interest has turned to understanding the inter-individual gene variability and how such variability affects the susceptibility of different individuals to disease and to drugs. In the field of public health, there is also a need for rapid screening of pathogens causing public health emergencies such as infectious diseases of unknown cause, all of which require rapid, reliable, economical and high throughput detection of DNA samples.
The gene detection plays a significant role in diagnosis and treatment, prognosis prediction, medication guidance and the like of hereditary diseases, tumors and infectious diseases. The current detection methods are numerous and distinctive. The detection means based on PCR, such as ARMS-PCR, fluorescent quantitative PCR, digital PCR and the like, has simple and convenient operation, high speed and simple data analysis, but cannot meet the clinical requirement of detecting a plurality of genes and a plurality of loci due to the limitation of flux.
The current detection method based on fluorescence detection is the most extensive, but because of spectral overlap among fluorescence of each color, the flux of detection is fundamentally limited. Recently, fluorescence-encoded microsphere technology has been developed to address this problem by preparing large quantities of fluorescence-encoded microspheres. However, the preparation process is complex and difficult, and further coating may affect the detection of fluorescent signals during the use of fluorescently encoded microspheres. There is therefore an interest in exploring non-optical labels and detection methods that would simplify the labeling process and provide high throughput, rapid and accurate assays.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention provides a nucleic acid detection method based on a metal coding technology.
The purpose of the invention is realized by the following technical scheme:
the nucleic acid detection method based on the metal coding technology comprises the following steps:
encoding magnetic beads by using metal, and coupling a probe of a target gene with the magnetic beads to obtain metal-encoded probe magnetic beads;
extracting genes, and amplifying by using a primer modified with biotin;
co-incubating the metal-encoded probe magnetic beads and the amplified gene;
adding metal-labeled avidin to combine with the biotin, removing redundant components, and then sending to a mass spectrometer for detection; the metal labeling the avidin is different from the metal encoding the magnetic beads;
obtaining the content of metal ions in the avidin by using a mass spectrometry technology;
and obtaining the information of the gene by using the obtained metal ion content and the mapping relation, wherein the mapping relation is the corresponding relation between the metal ion content and the information of the gene.
Compared with the prior art, the invention has the beneficial effects that:
1. the process is simple and the difficulty is low;
mature technologies are utilized for gene extraction, preparation of probe magnetic beads, incubation and the like, so that complexity and difficulty are reduced;
2. the over-detection accuracy and the resolution are high;
in debugging, the spatial positions of the vertical torch tube and the coil are accurately and synchronously adjusted to reach the optimal position under each working condition, and the most accurate detection data is obtained;
the fine adjustment of the torch tube in three dimensions is realized, the position precision and the repetition precision in three directions can reach 0.01mm, and the optimal position of the flame and the cone of the torch tube is realized;
the flight time mass spectrometer can realize second-order time focusing on wider ion initial position dispersion, and the mass resolution is obviously improved;
3. the sensitivity is high;
the technical requirement on high-voltage pulse can be reduced by adopting a double-pulse repulsion technology; the invention adopts a double-repulsion mode of positive pulse pushing (repulsion electrode) and negative pulse pulling (traction electrode), the requirement of high voltage can be reduced by half, so that the rising edge is steeper and the pulse waveform can be improved;
the first grid and the second grid with equal electric potential are added in the middle of the double-pulse repulsion, so that the electric field permeation effect of the acceleration region on the ion modulation region can be reduced;
the first grid mesh and the second grid mesh are directly grounded, no extra voltage is added, and the adjusting difficulty is small;
the wider modulation region can be realized, and the ion flux and the sensitivity are improved;
4. the reliability is good;
the torch tube is vertically arranged and keeps relative static with the coil, and the torch tube and the coil move synchronously, so that the torch tube is prevented from being burnt out due to the fact that the torch tube is close to the coil when moving, and the problem that a sampling cone is burnt and deformed is solved;
the rotation of the motor is reliably converted into the vertical movement of the conversion piece by utilizing the conversion module and the conversion piece, and the bearing piece and the conversion piece only move in the vertical direction by using a plurality of guide pieces, so that the inclination of the bearing piece is effectively prevented, and the positioning accuracy and reliability of the torch tube are ensured;
drawings
The disclosure of the present invention will become more readily understood with reference to the accompanying drawings. As is readily understood by those skilled in the art: these drawings are only for illustrating the technical solutions of the present invention and are not intended to limit the scope of the present invention. In the figure:
FIG. 1 is a flow chart of a nucleic acid detection method based on metal encoding techniques according to an embodiment of the present invention;
FIG. 2 is a schematic diagram of a mass spectrometer according to an embodiment of the invention;
FIG. 3 is a schematic structural diagram of a carrying unit according to an embodiment of the present invention;
FIG. 4 is a schematic diagram of a time-of-flight mass analyzer in accordance with an embodiment of the present invention;
fig. 5 is a schematic structural diagram of a time-of-flight mass analyzer according to an embodiment of the present invention.
Detailed Description
Fig. 1-5 and the following description depict alternative embodiments of the invention to teach those skilled in the art how to make and use the invention. Some conventional aspects have been simplified or omitted for the purpose of explaining the technical solution of the present invention. Those skilled in the art will appreciate that variations or substitutions from these embodiments will be within the scope of the invention. Those skilled in the art will appreciate that the features described below can be combined in various ways to form multiple variations of the invention. Thus, the present invention is not limited to the following alternative embodiments, but is only limited by the claims and their equivalents.
Example 1:
fig. 1 is a flow chart of a nucleic acid detection method based on a metal coding technique according to an embodiment of the present invention, and as shown in fig. 1, the nucleic acid detection method based on the metal coding technique is:
encoding magnetic beads by using metal, and coupling a probe of a target gene with the magnetic beads to obtain metal-encoded probe magnetic beads;
extracting genes, and amplifying by using a primer modified with biotin;
co-incubating the metal-encoded probe magnetic beads and the amplified gene;
adding metal-labeled avidin to combine with the biotin, removing redundant components, and then sending to a mass spectrometer for detection; the metal labeling the avidin is different from the metal encoding the magnetic beads;
obtaining the content of metal ions in the avidin by using a mass spectrometry technology;
and obtaining the information of the gene by using the obtained metal ion content and the mapping relation, wherein the mapping relation is the corresponding relation between the metal ion content and the information of the gene.
In order to adapt to the simultaneous detection of various target genes, further, the mode of using metal-coded magnetic beads is as follows:
the metal ion chelate or the combination of a plurality of different metal ion chelates are respectively combined with different magnetic beads in a reaction way; removing the excess metal ion chelate;
adding a chemical activation reagent, activating surface groups of different magnetic beads, adding a modified nucleic acid probe, and incubating, wherein the nucleic acid probe comprises a multi-site probe aiming at a single gene and a probe combination for multi-gene detection; removing excessive target gene probe.
In order to reduce the complexity and difficulty of extracting genes, further, the extraction mode of the genes is as follows:
and extracting total RNA by using an RNA extraction kit, and carrying out reverse transcription on the total RNA by using a reverse transcription kit to obtain cDNA.
In order to accurately obtain the mapping relationship, further, the obtaining manner of the mapping relationship is as follows:
preparing a nucleic acid standard with gradient concentration;
and detecting the nucleic acid standard substance by using the detection method to obtain the metal ion content, thereby obtaining the corresponding relation between the metal ion content and the nucleic acid concentration.
In order to adapt to the detection of multiple target genes, the nucleic acid standard is further multiple, and the concentration of the multiple nucleic acid standards in each gradient concentration is the same.
In order to improve the detection accuracy, further, in the mass spectrometry, the specific ionization mode is as follows:
the motor rotates, and the conversion module converts the rotation of the motor into the linear movement of the sliding piece;
converting the linear movement of the sliding part into the vertical movement of the conversion part, so as to drive the bearing part connected with the conversion part to vertically move along a plurality of guide parts, and the bearing unit arranged on the bearing part vertically moves along with the bearing part; the torch tube is vertically arranged on the carrying unit, and the coil is fixed on the carrying unit and is kept static relative to the torch tube;
the position of the bearing unit is adjusted in a two-dimensional mode in the horizontal direction, the bearing unit is arranged on the two-dimensional adjusting unit, and the two-dimensional adjusting unit is arranged on the bearing piece;
the sample suspension enters the torch tube and is excited into plasma through the coil, so that ions are formed;
the ions pass through a sampling cone and enter a mass spectrometry unit.
In order to improve the detection accuracy, further, in the mass spectrometry, a time-of-flight mass analyzer is used, the time-of-flight mass analyzer comprises a repulsion electrode, a field-free flight area and a detector, and the field-free flight area comprises a first incidence grid; the time-of-flight mass analyser further comprises:
a first ion acceleration region is formed between the traction electrode and the first incident grid;
the device comprises a first grid and a second grid, wherein the potential difference between the first grid and the second grid is zero; a second ion acceleration area is formed between the repulsion electrode and the first grid, and between the second grid and the traction electrode; the ions sequentially pass through the first grid mesh, the second grid mesh, the traction electrode, the first incidence grid mesh and the field-free flight area and are received by the detector.
The time-of-flight mass analyser further comprises:
a reflective region including a first reflective field including a second incident grid and reflective electrodes and a second reflective field including the reflective electrodes and reflective plates; ions emerging from the field-free flight zone are reflected by the reflective zone and are then received by the detector.
In order to realize second-order focusing, the second ion acceleration region and the first and second reflection fields satisfy the following conditions:
E1、E3、E4、E5electric field intensity, z, of the second ion acceleration region, the first reflection field and the second reflection field, respectively0、dG、d2、d3、d4、d5Respectively the distance between the incident ion and the first grid, firstThe distance between the grid mesh and the second grid mesh, the distance between the second grid mesh and the traction electrode, the distance between the traction electrode and the first incident grid mesh, the distance between the second incident grid mesh and the reflecting electrode, and the distance between the reflecting electrode and the reflecting plate; l is the length of flight of the ions in the field-free region between the first entrance grid and the detector.
In order to realize second-order focusing, the second ion acceleration area and the field-free reflection area satisfy the following conditions:
E1、E3electric field intensity, z, of the second ion acceleration region and the first ion acceleration region, respectively0、dG、d2、d3The distance between the incident ions and the first grid, the distance between the first grid and the second grid, the distance between the second grid and the traction electrode, and the distance between the traction electrode and the first incident grid are respectively; l is the length of flight of the ions in the field-free region between the first entrance grid and the detector.
Example 2:
the nucleic acid detection method based on the metal coding technology in the embodiment 1 of the invention is an application example of HK2, PFK, PK, LDHA, GLUT1, PEPCK, G6Pase, FBP, PC, SIRT1 and PGC-1 alpha gene in simultaneous detection.
In this application example, the nucleic acid detection method based on the metal coding technique:
sample preparation: extracting total RNA by using an RNA extraction kit, and then carrying out reverse transcription on the total RNA by using a reverse transcription kit to obtain cDNA;
metal-encoded probe bead preparation: and incubating Pd salt solutions with the mass numbers of 102, 104, 105, 106, 108 and 110 with an intermediate to form metal ion chelates, reacting and combining the metal ion chelates with different magnetic beads according to a table combination, and removing the unreacted metal ion chelates by using magnetic adsorption.
| Gene | Combination of |
| |
102、104、105 |
| |
102、104、106 |
| |
102、104、108 |
| |
102、104、110 |
| |
102、105、106 |
| |
102、105、108 |
| G6Pase | 102、105、110 |
| FBP | 102、106、108 |
| |
102、106、110 |
| |
102、108、110 |
| PGC-1α | 104、105、106 |
Adding a chemical activation reagent on the basis of metal-coded magnetic beads, activating groups on the surfaces of the magnetic beads, then adding a specially modified target gene probe, and removing redundant unreacted target gene probes by using magnetic adsorption after incubation;
adding metal coded probe magnetic beads into a hybridization system containing target genes, uniformly mixing, and incubating at 37 ℃ for 30 min;
placing the sample on a magnetic frame, separating magnetic beads, and removing a supernatant;
adding corresponding washing buffer solution, and washing twice;
adding metal-labeled avidin, mixing, and incubating at 37 deg.C for 10 min;
placing the sample on a magnetic frame, separating magnetic beads, and removing a supernatant;
adding corresponding washing buffer solution, and washing twice;
resuspending a metal-coded probe magnetic bead-target gene-metal-marked avidin complex, delivering the heavy suspension to ICP-TOF-MS, firstly ionizing to release metal ions and further delivering the released metal ions into a time-of-flight detection chamber for detection, and further delivering the released labeled metal ions into a time-of-flight mass analyzer for detection; the metal ions marked on the magnetic beads can distinguish different types of gene indexes to be detected, and the content of the metal ions marked on the avidin can indirectly reflect the expression quantity of each target gene; the detector will accurately record the exact content of the isotope designed and converted to various labels and analyze the output data by means of the adaptation software (mapping relationship between metal ion and gene concentration).
The obtaining relationship of the mapping relationship is as follows:
preparing HK2, PFK, PK, LDHA, GLUT1, PEPCK, G6Pase, FBP, PC, SIRT1 and PGC-1 alpha standard products with gradient concentration, wherein the concentration of each gradient concentration of the plurality of nucleic acid standard products is the same;
detecting HK2, PFK, PK, LDHA, GLUT1, PEPCK, G6Pase, FBP, PC, SIRT1 and PGC-1 alpha with gradient concentration by using the metal-coded probe magnetic beads and the metal-labeled avidin in the detection method and ICP-TOF-MS, and respectively obtaining the content of metal ions in the corresponding avidin so as to obtain the mapping relation between the concentration gradient and the content of the metal ions;
the specific structure of ICP-TOF-MS is as follows:
as shown in fig. 2, the torch tube 101 and the coil 102 are respectively arranged on different parts of the carrier unit, the torch tube 101 and the coil 102 remaining relatively stationary;
as shown in fig. 3, the carrying unit includes a fixing portion 301, a first mounting portion 302, a second mounting portion 303, and a connecting portion 304; the second mounting part 303 is horizontally fixed on the first adjusting unit 201, the fixing part 301 is fixed on the upper side of the second mounting part 303, the connecting part 304 is vertically arranged, the lower end is fixed on the second mounting part 303, and the upper end is fixed with the horizontally arranged first mounting part 302; the torch tube 101 is vertically arranged on the first mounting part 302, one end of the coil 102 is fixed on the fixing part 301, and the other end of the coil surrounds the torch tube 101;
as shown in fig. 2, the second adjusting unit is fixed on the chassis 100 and comprises a motor 401, a screw 402, a nut, a guide rail 404, a slider 405 and a bearing, wherein the motor 401 drives the horizontally arranged screw 402 to rotate, the nut is sleeved on the screw 402 by using a thread, the slider 405 is arranged on the guide rail 404 arranged in parallel with the screw 402, and the nut is connected with the slider 405, so that when the unit screw 402 rotates, the slider 405 is driven to horizontally and linearly move along the guide rail 404; the bearing is disposed at the top end of the slider 405; the four corners of the carrier 503 are respectively provided with through holes for allowing the vertically arranged guides 504 to pass through, and the difference between the inner diameter of the through holes and the outer diameter of the guides 504 is small, so that the carrier 503 can only move vertically along the guides 504; the conversion piece is provided with a vertical part 501 and an inclined part 502, the lower end of the vertical part 501 is connected with the bearing part 503, the upper end of the vertical part is connected with the inclined part 502, the inclined part 502 is propped by the bearing, and the sliding piece 405 moves horizontally and linearly on the lower side of the inclined part 502, so that the inclined part 502 only moves vertically;
the first adjusting unit 201 adopts an electric two-dimensional moving platform to realize two-dimensional adjustment in the horizontal direction, and is arranged on the bearing part 503;
fig. 4 is a schematic structural diagram of a time-of-flight mass analyzer according to an embodiment of the present invention, and as shown in fig. 4, the time-of-flight mass analyzer includes:
a repeller 11, a field-free flight zone 30 and a detector 51, said field-free flight zone 30 comprising a first entrance grid 31;
a first ion acceleration region is formed between the traction electrode 12 and the first incident grid 31;
a first grid 21 and a second grid 22, wherein the potential difference between the first grid 21 and the second grid 22 is zero; a second ion acceleration region is formed between the repulsion electrode 11 and the first grid 21, and between the second grid 22 and the traction electrode 12; the traction electrode is provided with a slotted hole allowing ions to pass through or a grid mesh structure allowing ions to pass through; the ions sequentially pass through the first grid 21, the second grid 22, the traction electrode 12, the first incidence grid 31 and the field-free flight area 30, and are received by the detector 51; the first grid 21 and the second grid 22 are grounded, so that the first grid 21 and the second grid 22 are ensured to be in equal potential;
a plurality of electrodes allowing ions to pass through are arranged between the traction electrode 12 and the first incidence grid 31, and voltage division is carried out on the plurality of electrodes by using a voltage division resistor; a plurality of electrodes allowing ions to pass through are arranged between the traction electrode 12 and the first incident grid 31, and the voltage of the plurality of electrodes is divided by using a voltage dividing resistor, so that the electric field intensity of the first ion acceleration area is uniform; the power supply applies a positive pulse voltage to the repeller 11 and a negative pulse voltage to the trailing electrode 12.
In order to realize second-order focusing, the distance between the first grid and the second grid and the field-free flight area satisfy that:
E1、E3electric field intensity, z, of the second ion acceleration region and the first ion acceleration region, respectively0、dG、d2、d3The distance between the incident ion and the first grid 21, the distance between the first grid 21 and the second grid 22, the distance between the second grid 21 and the traction electrode 12, and the distance between the traction electrode 12 and the first incident grid 31, respectively; l is the length of flight of ions between the first entrance grid 31 and the detector 51 in the field-free region.
The ionization mode of the single cell is as follows:
the motor rotates to drive the nut to carry the sliding piece 405 to move horizontally and linearly on the guide rail 404;
the sliding member 405 moves linearly horizontally under the inclined portion 502 of the converting member, thereby converting into a vertical movement of the converting member (the converting member only moves vertically), thereby driving the carrier 503 connected to the converting member to move vertically along the plurality of guides 504 (the carrier 503 only moves vertically), and the carrier unit disposed on the carrier 503 moves vertically along with the carrier 503; the torch tube 101 is vertically arranged on the first mounting portion 302 of the carrier unit, and the coil 102 is fixed on the fixing portion 301 of the carrier unit and is kept relatively stationary with respect to the torch tube 101;
two-dimensionally adjusting the position of the bearing unit in the horizontal direction by using a first adjusting unit 201, the bearing unit being disposed on the first adjusting unit 201, the first adjusting unit 201 being disposed on the bearing 503; it can be seen that in the three-dimensional adjustment of the carrier unit, the torch tube 101 and coil 102 remain relatively stationary;
the sample is excited into a plasma by the coil 102, forming ions, upon entering the torch 101;
the ions pass through a sampling cone 103 and enter the mass analysis unit.
Example 3:
the nucleic acid detection method based on the metal coding technology in the embodiment 1 of the invention is an application example of HK2, PFK, PK, LDHA, GLUT1, PEPCK, G6Pase, FBP, PC, SIRT1 and PGC-1 alpha gene in simultaneous detection.
In the time-of-flight mass analyzer, as shown in fig. 5, the first grid 21 and the second grid 22 are grounded, so that the first grid 21 and the second grid 22 are equal in potential; a plurality of electrodes allowing ions to pass through are arranged between the traction electrode 12 and the first incident grid 31, and the voltage of the plurality of electrodes is divided by using a voltage dividing resistor, so that the electric field intensity of the first ion acceleration area is uniform; the power supply applies positive pulse voltage to the repulsion electrode 11 and applies negative pulse voltage to the traction electrode 12;
the reflective region includes a first reflected field including the second incident grid 32 and the reflective electrode 41, and a second reflected field including the reflective electrode 41 and the reflective plate 42; ions exiting the field-free flight zone 30 are reflected by the reflecting zone and then received by the detector 51; the reflective electrode 41 has a slot hole for allowing ions to pass through, or has a grid structure for allowing ions to pass through;
arranging a plurality of electrodes allowing ions to pass through in the first ion acceleration area, the first reflection field and the second reflection field, and dividing the voltage of the plurality of electrodes by using a voltage dividing resistor so that the electric field intensity in the first ion acceleration area, the first reflection field and the second reflection field is uniform;
in order to realize second-order focusing, the second ion acceleration region and the first and second reflection fields satisfy the following conditions:
E1、E3、E4、E5the second ion acceleration region and the first ion acceleration regionElectric field strength of the zone, the first reflected field and the second reflected field, z0、dG、d2、d3、d4、d5The distance between the incident ion and the first grid 21, the distance between the first grid 21 and the second grid 22, the distance between the second grid 22 and the traction electrode 12, the distance between the traction electrode 12 and the first incident grid 31, the distance between the second incident grid 32 and the reflective electrode 41, and the distance between the reflective electrode 41 and the reflective plate 42, respectively; l is the length of flight of the ions in the field-free region between the first entrance grid 31 and the detector 51.
In ICP, unlike example 2, is:
1. the second mounting part and the connecting part are not arranged, the fixing part is directly fixed on the first adjusting unit, and the first mounting part is horizontally fixed on the fixing part;
2. the screw rod and the guide rail are kept parallel and are obliquely arranged relative to the horizontal plane;
3. the conversion piece comprises a horizontal part and a vertical part, and the horizontal part is supported by the sliding piece; when the slide member, which is moved in a straight line in an inclined manner, moves on the lower side of the horizontal portion of the switching member, the switching member moves vertically therewith, and only moves vertically.
Claims (10)
1. The nucleic acid detection method based on the metal coding technology comprises the following steps:
encoding magnetic beads by using metal, and coupling a probe of a target gene with the magnetic beads to obtain metal-encoded probe magnetic beads;
extracting genes, and amplifying by using a primer modified with biotin;
co-incubating the metal-encoded probe magnetic beads and the amplified gene;
adding metal-labeled avidin to combine with the biotin, removing redundant components, and then sending to a mass spectrometer for detection; the metal labeling the avidin is different from the metal encoding the magnetic beads;
obtaining the content of metal ions in the avidin by using a mass spectrometry technology;
and obtaining the information of the gene by using the obtained metal ion content and the mapping relation, wherein the mapping relation is the corresponding relation between the metal ion content and the information of the gene.
2. The method for detecting nucleic acid based on metal coding technique of claim 1, wherein the metal-coded magnetic beads are used in a manner that:
incubating the intermediate with metal salt solutions with different mass numbers to form metal ion chelates;
the metal ion chelate or the combination of a plurality of different metal ion chelates are respectively combined with different magnetic beads in a reaction way; removing the excess metal ion chelate;
adding a chemical activation reagent, activating surface groups of different magnetic beads, adding a modified nucleic acid probe, and incubating, wherein the nucleic acid probe comprises a multi-site probe aiming at a single gene and a probe combination for multi-gene detection; removing excessive target gene probe.
3. The method for detecting nucleic acid based on metal coding technology of claim 1, wherein the gene is extracted by:
and extracting total RNA by using an RNA extraction kit, and carrying out reverse transcription on the total RNA by using a reverse transcription kit to obtain cDNA.
4. The method for detecting nucleic acid based on metal coding technique according to claim 1, wherein the mapping relationship is obtained by:
preparing a nucleic acid standard with gradient concentration;
and detecting the nucleic acid standard substance by using the detection method to obtain the metal ion content, thereby obtaining the corresponding relation between the metal ion content and the nucleic acid concentration.
5. The method of claim 4, wherein the nucleic acid standard is a plurality of nucleic acid standards, and the concentration of the plurality of nucleic acid standards is the same for each gradient concentration.
6. The method for detecting nucleic acid based on metal coding technology of claim 1, wherein the ionization in mass spectrometry is specifically as follows:
the motor rotates, and the conversion module converts the rotation of the motor into the linear movement of the sliding piece;
converting the linear movement of the sliding part into the vertical movement of the conversion part, so as to drive the bearing part connected with the conversion part to vertically move along a plurality of guide parts, and the bearing unit arranged on the bearing part vertically moves along with the bearing part; the torch tube is vertically arranged on the carrying unit, and the coil is fixed on the carrying unit and is kept static relative to the torch tube;
the position of the bearing unit is adjusted in a two-dimensional mode in the horizontal direction, the bearing unit is arranged on the two-dimensional adjusting unit, and the two-dimensional adjusting unit is arranged on the bearing piece;
the sample enters the torch tube and is excited into plasma through the coil, so that ions are formed;
the ions pass through a sampling cone and enter a mass spectrometry unit.
7. The metal-coding-technique-based nucleic acid detection method of claim 1, wherein in mass spectrometry, a time-of-flight mass analyzer is used, the time-of-flight mass analyzer comprising a repeller, a field-free flight zone and a detector, the field-free flight zone comprising a first entrance grid; the time-of-flight mass analyser further comprises:
a first ion acceleration region is formed between the traction electrode and the first incident grid;
the device comprises a first grid and a second grid, wherein the potential difference between the first grid and the second grid is zero; a second ion acceleration area is formed between the repulsion electrode and the first grid, and between the second grid and the traction electrode; the ions sequentially pass through the first grid mesh, the second grid mesh, the traction electrode, the first incidence grid mesh and the field-free flight area and are received by the detector.
8. The metal-coding-technique-based nucleic acid detection method of claim 7, wherein the time-of-flight mass analyzer further comprises:
a reflective region including a first reflective field including a second incident grid and reflective electrodes and a second reflective field including the reflective electrodes and reflective plates; ions emerging from the field-free flight zone are reflected by the reflective zone and are then received by the detector.
9. The method for detecting nucleic acid based on metal coding technology of claim 8, wherein the second ion acceleration region and the first and second reflection fields satisfy:
E1、E3、E4、E5electric field intensity, z, of the second ion acceleration region, the first reflection field and the second reflection field, respectively0、dG、d2、d3、d4、d5The distance between the incident ions and the first grid, the distance between the first grid and the second grid, the distance between the second grid and the traction electrode, the distance between the traction electrode and the first incident grid, the distance between the second incident grid and the reflection electrode, and the distance between the reflection electrode and the reflection plate are respectively; l is the length of flight of the ions in the field-free region between the first entrance grid and the detector.
10. The method for detecting nucleic acid based on metal coding technology of claim 7, wherein the distance between the first grid and the second grid and the field-free flight area satisfy the following conditions:
E1、E3electric field intensity, z, of the second ion acceleration region and the first ion acceleration region, respectively0、dG、d2、d3The distance between the incident ions and the first grid, the distance between the first grid and the second grid, the distance between the second grid and the traction electrode, and the distance between the traction electrode and the first incident grid are respectively; l is the length of flight of the ions in the field-free region between the first entrance grid and the detector.
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