WO2021201635A1 - Système et procédé d'analyse - Google Patents

Système et procédé d'analyse Download PDF

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Publication number
WO2021201635A1
WO2021201635A1 PCT/KR2021/004085 KR2021004085W WO2021201635A1 WO 2021201635 A1 WO2021201635 A1 WO 2021201635A1 KR 2021004085 W KR2021004085 W KR 2021004085W WO 2021201635 A1 WO2021201635 A1 WO 2021201635A1
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Prior art keywords
electromagnet unit
electromagnet
analyte
magnetic field
unit
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PCT/KR2021/004085
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English (en)
Korean (ko)
Inventor
강민희
김은주
김한비
최영만
Original Assignee
사회복지법인 삼성생명공익재단
아주대학교 산학협력단
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Publication of WO2021201635A1 publication Critical patent/WO2021201635A1/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/38—Diluting, dispersing or mixing samples
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/65—Raman scattering
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor

Definitions

  • the present invention relates to a separation apparatus and method for separating an analyte and an interfering material from a complex sample, and to an analysis system and method for detecting a target gene and confirming the presence or absence of a specific disease using the same.
  • a method of detecting a nucleic acid uses a labeled detection method using a second probe labeled with fluorescence, which allows a target nucleic acid to complementarily bind to a first probe for a probe and confirm it.
  • a detection method is easy to analyze a trace amount of nucleic acid, it is necessary to use different sensor chips to which a probe is immobilized, and a nucleic acid amplification technique such as a PCR amplification method must be preceded. ) may occur.
  • the method using a fluorescently labeled probe in PCR is one of the methods to detect only a specific amplification product using the 5' nuclease activity of Taq DNA polymerase.
  • Loop-mediated isothermal amplification is a simple and fast isothermal PCR. Instead of using Taq DNA polymerase used in existing PCR methods, Bst DNA with exonuclease function This is a method using a polymerase (Bst DNA polymerase). Since this isothermal PCR method does not require a change in temperature during gene amplification, it enables gene amplification at a fixed temperature easily without specialized equipment.
  • An object of the present invention is to provide a separation device capable of separating interfering substances other than analytes by using an electromagnet, and it is also an object of the present invention to provide an analysis system capable of integrating sample amplification, analyte separation and detection is to provide
  • the analysis system includes a separation device for agitating and separating a sample including an analyte, an interference material, and magnetic particles bound to the interference material, and is connected to the separation device and provides light to the sample. and an analysis device for irradiating light and detecting the analyte using an emitted light signal, wherein the separation device includes a body, a receiving unit detachably disposed on the body, and a receiving unit for accommodating the sample; A first electromagnet part disposed on the body and generating a magnetic field, and a second electron disposed to face the first electromagnet part around the receiving part on the body, and generating a magnetic field in a different direction from that of the first electromagnet part and a magnet, wherein at least one of the first electromagnet unit and the second electromagnet unit applies a magnetic field to the magnetic particles to agitate at least a portion of the analyte, the interference material, and the magnetic particles.
  • the first electromagnet unit and the second electromagnet unit may alternately apply a magnetic field to the magnetic particles at a preset period.
  • the first electromagnet unit and the second electromagnet unit may simultaneously apply a magnetic field to the magnetic particles at a preset period.
  • At least one of the first electromagnet unit and the second electromagnet unit may separate the analyte and the interference material by continuously applying a magnetic field to the magnetic particles.
  • the separated interfering material is located in a first region in the accommodating part, and the separated analyte is located in a second region different from the first region in the accommodating part, and the first The region may be a region adjacent to at least one of the first electromagnet unit and the second electromagnet unit, and the second region may be a region spaced apart from the first region.
  • an incident hole for guiding the light to the second region may be formed in the body of the separation device.
  • the optical signal may be a surface-enhanced Raman scattering (SERS) signal.
  • SERS surface-enhanced Raman scattering
  • the receiving part may be made of a transparent material.
  • the present invention is disposed on the body, and at least one additional electromagnet unit for generating a magnetic field in a different direction from the first electromagnet unit and the second electromagnet unit; further comprising, the additional electromagnet unit
  • a plurality of the additional electromagnet parts may be provided, and the plurality of additional electromagnet parts may be disposed to be spaced apart from the first electromagnet part and the second electromagnet part.
  • the analysis method comprises the steps of: preparing a sample containing an analyte, an interfering material, and magnetic particles in a receiving unit; amplifying the analyte and the interfering material; Separating the interfering material, irradiating light to the sample, and detecting the analyte using an emitted light signal, wherein separating the analyte and the interfering material includes the receiving generating a magnetic field by operating at least one of the first electromagnet unit and the second electromagnet unit disposed to face each other around the unit; It may include agitating the sample so that it binds, and separating the analyte and the interfering material in the receiving unit using the magnetic field.
  • the step of agitating the sample so that the magnetic particles bind to any one of the analyte or the interference material accommodated in the receiver using the magnetic field, the first electromagnet and the second electromagnet may include applying a magnetic field to the magnetic particles alternately at a preset period.
  • the step of stirring the sample so that the magnetic particles bind to any one of the analyte or the interfering material accommodated in the receiving unit using the magnetic field comprises: the first electromagnet unit and the second The electromagnet unit may simultaneously apply a magnetic field to the magnetic particles at a preset period.
  • At least one of the first electromagnet unit and the second electromagnet unit is the magnetic particle
  • the separated interference material is located in a first region in the accommodating part, the separated analyte is located in a second region in the accommodating part, and the first region is the first electron
  • the region may be adjacent to at least one of the magnet and the second electromagnet, and the second region may be a region spaced apart from the first region.
  • the light in the detecting of the analyte using the optical signal, the light may be guided to the second region through an incident hole formed in the body of the separation device.
  • the optical signal may be a surface-enhanced Raman scattering signal.
  • the receiving part may be made of a transparent material.
  • the present invention is disposed on the body, and at least one additional electromagnet unit for generating a magnetic field in a different direction from the first electromagnet unit and the second electromagnet unit; further comprising, the additional electromagnet unit
  • a plurality of the additional electromagnet parts may be provided, and the plurality of additional electromagnet parts may be disposed to be spaced apart from the first electromagnet part and the second electromagnet part.
  • the separation apparatus and separation method according to the embodiments of the present invention bind magnetic particles to an interference material using probe materials that specifically bind to an analyte and an interference material, respectively, and magnetic particles by means of symmetrically arranged electromagnets.
  • a magnetic field to the accommodating part to agitate the sample, it is possible to facilitate the binding of the magnetic particles and the probe.
  • the separation process and time can be shortened by separating the analyte and the interfering material directly in the accommodating part using the magnetic field generated by the symmetrically arranged electromagnets.
  • the analyte is separated from the interfering material in the receiving unit using a magnetic field, and then the analyte is performed by the analysis device without a separate process of discharging the interfering material.
  • FIG. 1 is a perspective view showing a separation device according to an embodiment of the present invention.
  • Figure 2 is an exploded view showing the separation device of Figure 2;
  • FIG. 3 is a schematic cross-sectional view of the separation device of FIG. 1 .
  • FIG. 4 is a graph showing the intensity and period of a magnetic field applied to magnetic particles.
  • FIG. 5 shows a stirring process using the separation device of FIG. 1 .
  • FIG. 6 illustrates a separation process using the separation device of FIG. 1 .
  • FIG. 7 is a flowchart illustrating a sequence of stirring and separating a sample according to an embodiment of the present invention.
  • FIG. 8 is a perspective view illustrating an analysis system according to an embodiment of the present invention.
  • FIG. 9 is a graph showing a surface-enhanced Raman scattering analysis signal of an analyte according to an embodiment of the present invention.
  • FIG. 10 is a flowchart illustrating an analysis procedure of a sample using the analysis system according to an embodiment of the present invention.
  • FIG. 1 is a perspective view showing a separation device according to an embodiment of the present invention.
  • FIG. 2 is an exploded view illustrating the separation device of FIG. 2
  • FIG. 3 is a cross-sectional view schematically illustrating the separation device of FIG. 1 .
  • 4 is a graph showing the intensity and period of a magnetic field applied to magnetic particles.
  • FIG. 5 shows a stirring process using the separation device of FIG. 1
  • FIG. 6 shows a separation process using the separation device of FIG. 1 .
  • the separation device 10 may include a body 100 , an accommodating part 110 , a first electromagnet part 120 , and a second electromagnet part 130 .
  • the body 100 may have a space for accommodating the accommodating part 110 therein.
  • the body 100 may have various shapes such as a hemisphere, a cylinder, a polygon, etc., but for convenience of explanation, hereinafter, the body 100 will be described mainly in an embodiment in which the body 100 is a rectangular parallelepiped shape with a space formed therein.
  • the accommodating part 110 is disposed in the inner space of the body 100 , and a sample may be accommodated in the accommodating part 110 .
  • the sample may be obtained from a biological sample of the subject, for example, from a tissue extract, cell lysate, whole blood, plasma, serum, saliva, ocular fluid, cerebrospinal fluid, sweat, urine, milk, ascites fluid, synovial fluid, or peritoneal fluid. It may be a nucleic acid sample obtained, but is not limited thereto. In this case, the subject may be a mammal including a human.
  • the sample may include an analyte to be detected, an interfering material not to be detected, and a magnetic material binding to any one of the analyte or the interfering material, and the sample may include a probe material and a liquid.
  • the magnetic particles may bind to the analyte or to the interfering material, but for convenience of description, an embodiment in which the magnetic particles bind to the interfering material will be mainly described.
  • the analyte may be a reporter material for diagnosing a specific disease
  • the interfering material may be a material other than the analyte included in the sample.
  • the probe material may include a first probe and a second probe that specifically bind to a target gene (infectious disease-specific gene).
  • the first probe is for detection of a target gene
  • a reporter material may be bound to the first probe
  • the reporter material may be a fluorophore or a Raman active material. Since the fluorescent material or Raman active material shows a specific spectrum, a target material to be detected can be analyzed more effectively.
  • Fluorescent material or Raman active material capable of binding to the first probe is, for example, FAM (5'-Fluorescein phosphoramidite), TAMRA (5'-TAMRA phosphoramidite), VIC (2'-chloro-7''phenyl-1) ,4-dichloro-6-carboxy-fluorescein), Cy5(2-[5-[1,3-Dihydro-1-[3-(4-methoxytriphenylmethoxy)propyl]-3, 3-dimethyl-2H-indol-2) -ylidene]-1, 3-pentadien-1-yl]-1-[3-[N,N-diisopropylamino(2-cyanoethoxy)phosphinoxy]propyl]-3, 3-dimethyl-3H-indolium), JOE (6 -Carboxy-4', 5'-Dichloro-2', 7'-Dimethoxyfluorescein, Succinimidyl Ester),
  • the second probe is for separating substances other than the analyte, and the second probe may be a substance capable of specifically binding to a residue of a magnetic particle or a substance bound to the magnetic particle.
  • the magnetic particles may be, for example, magnetite (Magnetite; Fe 3 O 4 ), maghemite (Fe 2 O 3 ), or hematite (Hematite; Fe 2 O 3 ).
  • Magnetite may include r-Fe 2 O 3 in a core-shell structure, and r may be made of Si, ZnO, Fe, Co and Mg.
  • the present invention is not limited thereto, and any magnetic particles known in the art may be used as the magnetic particles.
  • the PCR amplification process may proceed.
  • the reporter material bound to the first probe in the PCR amplification product may be separated from the first probe by the restriction enzyme activated during the PCR amplification process.
  • the probe material (ie, the second probe) other than the first probe bound to the target gene is specifically bound to the magnetic particle, the magnetic particle may be bound to the interference material.
  • Separation device 10 for generating a magnetic field can be used to separate interfering substances.
  • the analyte (ie, the reporter material) from which the interfering material is separated may be detected by the analysis device 20 such as a Raman spectrometer.
  • the reporter material is not separated from the first probe, and thus the reporter material may remain bound to the probe material.
  • the second probe which is another material included in the probe material, is combined with the magnetic particles, the analyte (ie, the reporter material) may be separated by the separation device 10 together with the interfering material.
  • the analyte since the analyte is separated together by the separation device 10 in a state of being bound to the interference substance, the analyte may not be detected by the analysis device 20 .
  • the receiving part 110 may be disposed on the body 100 .
  • the accommodating part 110 is detachably connected to the body 100, and when the user needs to change the test target sample, the accommodating part 110 is separated from the body 100 and replaced with a new accommodating part 110.
  • the receiving unit 110 It is possible to prevent a decrease in analysis accuracy due to residues of existing experiments that may remain in the .
  • the receiving part 110 may be supported by the supporting part 101 of the body 100 .
  • the receiving part 110 may be fixed to the support part 101 after being inserted into the body 100 . Accordingly, it is possible to improve the stability of the analysis by preventing the receiving unit 110 from shaking due to vibration, shock, etc. that may occur in the process of performing analysis such as stirring and separation.
  • the accommodating part 110 may be made of a transparent material through which light can pass.
  • the receiving part 110 made of a transparent material may have a high transmittance. Thereby, the light emitted from the analysis device 20 can be incident into a target area in the separation device 10 . Details on this will be described later.
  • the first electromagnet unit 120 may generate a magnetic field by receiving a current from a current applying unit (not shown) disposed outside or inside the separation device 10 .
  • the first electromagnet unit 120 may be disposed on one side of the body 100 .
  • the first electromagnet part 120 may include a first base part 121 , a first electromagnet unit 122 , and a first protrusion part 123 .
  • the first base part 121 may be a part of one side surface of the body 100 on which the first electromagnet part 120 is disposed, and the first electromagnet unit 122 may be disposed on the first base part 121 . have.
  • the first electromagnet unit 122 may include an electromagnet having a plurality of coils C1 wound therein.
  • a current is applied to the first electromagnet unit 122 from a current applying unit (not shown)
  • the first electromagnet unit 122 may generate a magnetic field.
  • the direction and strength of the current applied to the first electromagnet unit 122 or the number and winding direction of the coil C1 can be adjusted. have.
  • the first protrusion 123 may be disposed between the first electromagnet unit 122 and the receiving part 110 inserted into the body 100 .
  • the first protrusion 123 may protrude from one end of the first electromagnet unit 122 toward the receiving unit 110 in a direction perpendicular to the radial direction of the first electromagnet unit 122 .
  • the first protrusion 123 may support the receiving unit 110 by contacting one side of the receiving unit 110 .
  • the first electromagnet unit 120 may apply a magnetic field generated by the first electromagnet unit 122 to the receiving unit 110 . Specifically, the first electromagnet unit 120 may apply a magnetic field to the sample (eg, magnetic particles) accommodated in the receiving unit 110 .
  • the second electromagnet unit 130 may generate a magnetic field by receiving a current from a current applying unit (not shown) disposed outside or inside the separation device 10 .
  • the second electromagnet unit 130 may be disposed on one side and the other side of the body 100 on which the first electromagnet unit 120 is disposed. In this case, the second electromagnet unit 130 may be disposed on the side of the body 100 on which the first electromagnet unit 120 is not disposed.
  • An embodiment that is disposed on the other side of the body 100 so as to face the first electromagnet unit 120 disposed on one side of the body 100 will be mainly described.
  • the second electromagnet unit 130 may include a second base unit 131 , a second electromagnet unit 132 , and a second protrusion 133 .
  • the second electromagnet unit 132 may include an electromagnet in which a plurality of coils C2 are wound.
  • the second electromagnet unit 132 may generate a magnetic field.
  • the direction and strength of the current applied to the second electromagnet unit 132 or the number and winding direction of the coil C2 can be adjusted. .
  • the second protrusion 133 may be disposed between the second electromagnet unit 132 and the receiving part 110 inserted into the body 100 .
  • the second protrusion 133 may protrude from one end of the second electromagnet unit 132 toward the receiving unit 110 in a direction perpendicular to the radial direction of the second electromagnet unit 132 .
  • the second protrusion 133 may be disposed to be symmetrical with the first protrusion 123 with the accommodating part 110 interposed therebetween to support the accommodating part 110 in both directions. Thereby, the stability of agitation and separation of the sample can be improved.
  • each component included in the second electromagnet unit 130 is the same as or similar to those of the first electromagnet unit 120 , and thus a redundant description thereof will be omitted.
  • the second electromagnet unit 130 may apply a magnetic field generated by the second electromagnet unit 132 to the receiving unit 110 . Specifically, the second electromagnet unit 130 may apply a magnetic field to the sample accommodated in the receiving unit 110 (eg, magnetic particles in the receiving unit 110).
  • At least one of the first electromagnet unit 120 and the second electromagnet unit 130 may apply a magnetic field to the magnetic particles accommodated in the receiving unit 110 . Accordingly, at least a portion of the analyte, the interfering material, and the magnetic particles may be stirred in the receiving unit 110 . In this case, the direction and strength of the magnetic field applied by the first electromagnet unit 120 and the magnetic field applied by the second electromagnet unit 130 may be different from each other. However, the present invention is not limited thereto, and the directions and strengths of the magnetic fields may be the same.
  • the direction of the magnetic field generated by the first electromagnet unit 120 and the second electromagnet unit 130 is the winding direction of the coils C1 and C2 wound on each electromagnet unit or the coils C1 and C2 as described above. It can be adjusted by changing the direction of the current applied to it.
  • Stirring by the magnetic field can be controlled by changing a predetermined condition.
  • the intensity of agitation may be adjusted by changing the intensity of the direct current applied to the first electromagnet unit 120 and the second electromagnet unit 130 and the number of turns of the coils C1 and C2.
  • the frequency of the alternating current applied to the first electromagnet unit 120 and the second electromagnet unit 130 and the duty cycle of the first electromagnet unit 120 and the second electromagnet unit 130 The cycle of agitation can be adjusted.
  • the stirring by the magnetic field may be adjusted to change over time.
  • the first electromagnet unit 120 and the second electromagnet unit 130 may apply a magnetic field to the magnetic particles at a preset period (T).
  • the preset period (T) is the first electromagnet unit 120 or the second electromagnet unit from a point in time when any one of the first electromagnet unit 120 or the second electromagnet unit 130 starts to apply a magnetic field to the magnetic particles.
  • Any one of 130 may mean a time before starting to apply a magnetic field to the magnetic particles again, and the preset period T includes a plurality of the first period T1, the second period T2, and the like. It can be repeated in cycles.
  • the first electromagnet unit 120 and the second electromagnet unit 130 may alternately apply a magnetic field to the magnetic particles.
  • any one of the first electromagnet unit 120 and the second electromagnet unit 130 may first apply a magnetic field to the magnetic particles for the first time t1 .
  • the other of the first electromagnet unit 120 and the second electromagnet unit 130 may apply a magnetic field to the magnetic particles for a second time t2.
  • the first electromagnet unit 120 applies a magnetic field to the magnetic particles for a first time t1, and then the second electromagnet unit 130 generates magnetic particles for a second time t2.
  • An embodiment in which a magnetic field is applied will be mainly described.
  • a first time t1 in which the first electromagnet unit 120 applies a magnetic field and a second time t2 in which the second electromagnet unit 130 applies a magnetic field may be continuous (refer to FIG. 4A ).
  • the magnetic particles by any of the first electromagnet unit 120 and the second electromagnet unit 130 may be an unattended application time (to) in which no magnetic field is applied (see FIG. 4B ).
  • the first time t1 and the second time t2 may be the same, but as another example, the first time t1 and the second time t2 may be different.
  • the unattended time to may be the same as or different from any one of the first time t1 and the second time t2.
  • the first electromagnet unit 120 and the second electromagnet unit 130 continuously alternate to apply a magnetic field to the magnetic particles so that there is no time when the magnetic field is not applied to the magnetic particles while the stirring is in progress. It can also be applied (see Fig. 4c).
  • any one of the first electromagnet unit 120 or the second electromagnet unit 130 continuously applies a magnetic field a plurality of times, one of the first electromagnet unit 120 or the second electromagnet unit 130 .
  • the other may apply a magnetic field (not shown).
  • the magnetic particles in the receiving unit 110 are generated by the magnetic field in the first electromagnet unit 120 ) and the movement toward the second electromagnet unit 130 may be repeated.
  • the movement of the magnetic particles stirs the liquid accommodated in the accommodating part to generate a flow of the liquid in the accommodating part 110 .
  • the analyte and the interfering material accommodated in the receiving unit 110 are uniformly mixed with the magnetic particles, so that the sample can be stirred in the receiving unit 110 .
  • the magnetic particles can be effectively bound to the interference substance (or the binding material bound to the interference substance), thereby increasing the amount of magnetic particles bound to the interference substance.
  • the first electromagnet unit 120 and the second electromagnet unit 130 may simultaneously apply a magnetic field to the magnetic particles at a preset period.
  • the first electromagnet unit 120 and the second electromagnet unit 130 may simultaneously apply a magnetic field to the magnetic particles for the same application time ts. For example, between the application time ts1 by the first electromagnet unit 120 and the second electromagnet unit 130 and the next application time ts2, there is an unattended time to in which a magnetic field is not applied to the magnetic particles. can be (see Figure 4d).
  • the magnetic particles before the stirring of the sample is started by the magnetic field, the magnetic particles may be in a state of sinking to the lower part of the inside of the receiving unit 110 . Thereafter, when a current is applied to the first electromagnet unit 120 and the second electromagnet unit 130 to apply a magnetic field to the magnetic particles, the magnetic particles are generated in the receiving unit 110 by the magnetic field in the first electromagnet unit 120 . By repeating the movement toward and movement toward the second electromagnet unit 130 , a flow may be formed in the liquid inside the receiving unit 110 . When the magnetic field is applied to the magnetic particles for a sufficient time and the application of the magnetic field is stopped after the stirring is progressed, the magnetic particles may be uniformly dispersed in the receiving unit 110 .
  • the first electromagnet unit 120 and the second electromagnet unit 130 may re-apply a magnetic field to the magnetic particles in the receiving unit 110 .
  • the first electromagnet unit 120 and the second electromagnet unit 130 continuously apply a magnetic field to the magnetic particles at the same time, whereby some of the magnetic particles are part of the first electromagnet unit in the receiving unit 110 by the magnetic field. It may move toward 120 and may be located on the periphery of the inner wall of the accommodating part 110 adjacent to the first electromagnet part 120 .
  • the other part of the magnetic particles moves toward the second electromagnet unit 130 within the receiving unit 110 by the magnetic field to be located in the inner wall periphery of the receiving unit 110 adjacent to the second electromagnet unit 130 .
  • the interference material to which the magnetic particles are bound and the analyte to which the magnetic particles are not bound may be separated into different regions (eg, the first region and the second region) inside the receiving unit 110 .
  • the first electromagnet unit 120 and the second electromagnet unit 130 simultaneously apply a magnetic field to the magnetic particles
  • the first electromagnet unit 120 and the second electromagnet unit 130 alternately generate a magnetic field.
  • a stronger force can be applied to the magnetic particles compared to the case of applying
  • the flow of the liquid in the accommodating part 110 is reduced.
  • the analyte and the interfering material in the accommodating unit 110 may be uniformly mixed with the magnetic particles. Accordingly, it is possible to facilitate the binding of the magnetic particles to the interference material.
  • the separation device 10 may further include at least one additional electromagnet (not shown).
  • the first electromagnet unit 120 , the second electromagnet unit 130 , and at least one additional electromagnet unit may be disposed on the body 100 , and the perimeter of the receiving unit 110 with the receiving unit 110 as the center Accordingly, they may be disposed to be spaced apart from each other.
  • the first electromagnet unit 120 , the second electromagnet unit 130 , and at least one additional electromagnet unit may generate a magnetic field, and may apply the generated magnetic field to the magnetic particles accommodated in the receiving unit 110 .
  • the first electromagnet unit 120 and the second electromagnet unit 130 are disposed to face each other with the receiving unit 110 as the center as described above, and in this case, the additional electromagnet unit The part may be disposed on a side opposite to one surface of the body 100 to which the analysis device 20 is coupled.
  • the method of applying the magnetic field to the magnetic particles by the first electromagnet unit 120 , the second electromagnet unit 130 , and at least one additional electromagnet unit is the same as the above-described first electromagnet unit 120 and the second electromagnet unit 130 . or similar, overlapping description thereof will be omitted.
  • a plurality of electromagnet units disposed to surround the accommodating unit 110 applies magnetic fields in various directions different from each other to the magnetic particles accommodated in the accommodating unit 110 . can do. Thereby, the effect of stirring and separation by the magnetic field can be improved.
  • At least one of the first electromagnet unit 120 and the second electromagnet unit 130 may apply a magnetic field to the sample to separate the analyte and the interfering material.
  • the first electromagnet unit 120 and the second electromagnet unit 130 may continuously apply a magnetic field to the magnetic particles in the receiving unit 110 after the sample is stirred by the magnetic field and the magnetic particles are coupled to the interference material. .
  • the continuous application of the magnetic field is performed with the first electromagnet unit 120 and the first electromagnet unit 120 from after the sample is stirred and the magnetic particles are bound to the interfering material until the separation of the analyte and the interfering material in the receiving unit 110 is completed.
  • This may mean that the second electromagnet unit 130 operates to generate a magnetic field, and a state of applying the magnetic field to the sample (particularly, magnetic particles) in the receiving unit 110 is maintained.
  • the first electromagnet unit 120 and the second electromagnet unit 130 may continuously apply a magnetic field to the magnetic particles at the same time. Only the interference material to which the magnetic particles are bound by the applied magnetic field may be moved in the receiving unit 110 . Specifically, a part of the interference material to which the magnetic particles are bonded is attracted toward the first electromagnet unit 120 , and another part of the interference material to which the magnetic particles are bonded is attracted toward the second electromagnet unit 130 , and magnetic particles Since the analyte to which is not bound is not affected by the magnetic field, positional movement by the magnetic field may not occur. Accordingly, the analyte and the interfering material may be separated in the receiving unit 110 .
  • the first region may be a region adjacent to at least one of the first electromagnet unit 120 and the second electromagnet unit 130
  • the second region may be a region spaced apart from the first region.
  • the first region may be an inner wall periphery of the accommodating part 110 adjacent to the first electromagnet part 120 and an inner wall periphery of the accommodating part 110 adjacent to the second electromagnet part 130 .
  • the second region may be a region between the peripheral portions of both inner walls of the accommodating part 110 , or may be an inlet 111 side region of the accommodating part 110 .
  • the interference of the interfering material in the detection step of the analyte is minimized and the accuracy of the analyte detection is improved.
  • the analysis process can be simplified and the time required for the analysis can be shortened.
  • either the first electromagnet unit 120 or the second electromagnet unit 130 is continuously applied to the magnetic particles in the receiving unit 110 to separate the analyte and the interference material to which the magnetic particles are combined. may do it
  • FIG. 7 is a flowchart illustrating a sequence of stirring and separating a sample according to an embodiment of the present invention.
  • a method for stirring and separating a sample according to an embodiment of the present invention may be as follows.
  • a user may prepare a sample including an analyte, an interfering material, and magnetic particles in the receiving unit (S100).
  • the sample may include a probe material (eg, a first probe and a second probe).
  • At least one of the first electromagnet unit 120 and the second electromagnet unit 130 may be operated to generate a magnetic field (S200).
  • a magnetic field S200
  • the first electromagnet unit 120 and the second electromagnet unit 130 may generate a magnetic field.
  • This magnetic field may be applied to the magnetic particles accommodated in the receiving unit (110).
  • a magnetic field may be used to stir the sample so that the magnetic particles bind to the interfering material (S300).
  • a current applying unit (not shown) alternately applies current to the first electromagnet unit 120 and the second electromagnet unit 130 , and accordingly, the first electromagnet unit 120 and the second electromagnet unit 130 . 130 may apply a magnetic field to the magnetic particles in the receiving unit 110 by alternating with each other.
  • the magnetic particles in the accommodating part 110 are generated by a magnetic field, thereby generating a flow of the liquid in the accommodating part 110, and the analytes, interfering substances and magnetic particles accommodated in the accommodating part 110 by this flow can be stirred. have. Accordingly, it may be easy for the magnetic particles to bind to the interference material.
  • the analyte and the interfering material may be separated in the receiving unit 110 by using a magnetic field (S400).
  • a magnetic field S400
  • the first electromagnet unit 120 and the second electromagnet unit 130 provide a magnetic field to the magnetic particles of the stirred sample. can be continuously approved. Accordingly, the interference material to which the magnetic particles are bound may move to a first region in the accommodating unit 110 , and the analyte to which the magnetic particles are not bound may move to a second region in the accommodating unit 110 . Accordingly, the analyte and the interfering material may be separated in the receiving unit 110 .
  • the probe materials eg, the first probe and the second probe
  • the probe materials that specifically bind to the analyte and the magnetic particle, respectively
  • the separation process and time can be shortened by directly separating the analyte and the interfering material in the accommodating unit 110 using the magnetic field generated by the symmetrically arranged electromagnets 120 and 130 .
  • FIG. 8 is a perspective view illustrating an analysis system according to an embodiment of the present invention.
  • FIG. 9 is a graph showing a surface-enhanced Raman scattering analysis signal of an analyte according to an embodiment of the present invention.
  • 10 is a flowchart illustrating an analysis procedure of a sample using the analysis system according to an embodiment of the present invention.
  • the analysis system 1 may include a separation device 10 and an analysis device 20 .
  • the separation device 10 may stir and separate the sample including the analyte, the interference material, and the magnetic particle as described above.
  • the separation device 10 may include the body 100 , the accommodating part 110 , the first electromagnet part 120 , and the second electromagnet part 130 .
  • an incident hole 102 for guiding the light emitted from the analysis device 20 to either the first region or the second region in the receiving unit 110 may be formed.
  • the incident hole 102 is the analysis device
  • the light emitted from the light source of (20) may be guided to the analyte located in the second region.
  • the accommodating unit 110 is made of a transparent material that allows light to pass therethrough, whereby the light emitted from the light source 21 may be irradiated with the analyte contained in the accommodating unit 110 .
  • the incident hole 102 may be disposed on one side of the body 100 of the separation device 10 .
  • one side of the body 100 may be one side of the body 100 facing the analysis device 20 .
  • the incident hole 102 is disposed on a straight line with the light source of the analysis device 20 , and light emitted from the light source may travel toward the analyte through the incident hole 102 .
  • the analysis device 20 may measure an optical signal to detect an analyte present in the sample.
  • the analysis apparatus 20 may include a light source unit 21 and an optical signal detection unit (not shown), and in this case, the light source unit 21 and the optical signal detection unit may be disposed in the analysis apparatus 20 .
  • the light emitted from the light source unit 21 may be irradiated to the sample accommodated in the receiving unit 110 of the separation device 10 .
  • the light may be irradiated to the analyte in the second region of the receiving unit 110 through the incident hole 102 of the separation device 10 .
  • the optical signal detector may measure an optical signal generated when the irradiated light is scattered by the sample.
  • the analysis device 20 may measure the optical signal by various methods such as fluorescence or Raman spectroscopy, infrared spectroscopy, etc., but hereinafter, the analysis device 20 detects an analyte using Raman spectroscopy will be mainly described. decide to do
  • Raman scattering refers to a phenomenon in which when light passes through a medium, the wavelength of light is changed, so that a part of the light deviates from the direction of travel and proceeds in a different direction.
  • Raman spectroscopy is useful for obtaining molecular level information by irradiating a short-wavelength laser to a sample and detecting scattered light, and Raman spectroscopy provides information on the vibrational state of molecules.
  • absorbed radiation is re-radiated at the same wavelength, where the energy difference between the re-radiated beams is a shift in wavelength between them, and the degree of this difference is It is measured as a unit of wavenumber (the reciprocal of wavelength) and expressed as a Raman shift (RS).
  • the analysis device 20 may detect an analyte by measuring a surface-enhanced Raman scattering (SERS) signal.
  • the Raman signal may be measured by separating an analyte (eg, a fluorescent material or a Raman active material) from the first probe accommodated in the receiving unit 110 .
  • the analyte can be detected by irradiating the light emitted from the light source unit 21 of the analysis device 20 to the analyte accommodated in the receiving unit 110 to obtain a Raman signal of a fluorescent material or a Raman active material.
  • a specific peak is observed from the measured Raman signal, it can be determined that the individual has a marker of an infectious disease, and accordingly, whether the individual is infected can be confirmed.
  • the analysis device 20 may specify and detect any one target gene desired to be detected.
  • the receiving unit 110 includes a first probe material (eg, a first probe and a second probe of the first probe material) specifically binding to a first target gene, a second target gene, A second probe material that specifically binds (eg, the first probe and the second probe of the second probe material) may be included.
  • the reporter material ie, the analyte
  • the interfering material is separated by the magnetic field.
  • the reporter material is not dropped from the first probe of the second probe material, and the reporter material of the second probe material is combined with the second probe material. It can be separated from the analyte by the magnetic particles and the magnetic field applied to the magnetic particles. Accordingly, it can be seen that the light signal by the reporter material of the second probe material gradually decreases (for example, refer to FIG. 9A ). According to this method, multiple diagnosis may be possible from a single light source according to the fluorescent material introduced into the first probe.
  • the analysis device 20 may be detachably connected to the separation device 10 .
  • the analysis device 20 when analyzing a sample stirred and separated by the separation device 10 , the analysis device 20 may be connected to the separation device 10 .
  • the analysis device 20 when the analysis of the sample is completed by the analysis device 20 , the analysis device 20 may be separated from the separation device 10 .
  • the present invention is not limited thereto, and the analysis device 20 may analyze the sample by irradiating light toward the sample in the separation device 10 while being spaced apart from the separation device 10 .
  • the analysis device 20 may further include a power switch 22 and a display unit 23 of the analysis device 20 .
  • the display unit 23 may further include displaying information on whether an analyte is present and a diagnosis result of a specific disease of the subject based on the sample analysis result. Based on the information displayed on the display unit 23, the user may check whether the object has a disease.
  • a method of analyzing a sample using the analysis system 1 may be as follows, and for convenience of explanation, the following will focus on an embodiment in which a target gene is present in a sample accommodated in the receiving unit 110 . to be explained as
  • a user may prepare a sample including an analyte, an interference material, and magnetic particles in the receiving unit 110 of the separation device 10 ( S10 ).
  • the sample may include a liquid and a probe material including the first probe and the second probe that specifically bind to the target gene.
  • An analyte may be bound to the first probe, and magnetic particles may be bound to the second probe.
  • the analyte and the interfering material included in the sample may be amplified ( S20 ).
  • the analyte and the interfering material may be amplified through the PCR amplification process as described above.
  • the present invention is not limited thereto, and the analyte and the interfering material may be amplified by an isothermal amplification method.
  • the temperature in the accommodating unit 110 is used to amplify the By maintaining a specific temperature, analytes and interfering substances can be amplified without the need for an additional heat supply.
  • an analyte and an interfering material may be separated using a magnetic field in the separation device 10 ( S30 ).
  • the specific method of stirring and separating the analyte and the interfering material in the receiving unit 110 is the same as described above, the separated interfering material is located in the first region in the receiving unit 110, and the separated analyte is It may be located in the second area in the receiving unit 110 .
  • the analysis device 20 may measure the optical signal to detect the analyte present in the receiving unit 110 ( S40 ).
  • the analysis device 20 connected to the separation device 10 may irradiate the light emitted from the light source unit 21 as an analyte through the incident hole 102 , and measure a light signal of the light scattered by the analyte.
  • the optical signal may be a surface-enhanced Raman scattering signal, and by observing a specific peak from the measured Raman signal, it is possible to determine whether an individual is infected with a disease.
  • the analysis system 1 and the analysis method according to the embodiments of the present invention separate the analyte and the interfering material in the receiving unit 110 using a magnetic field and then discharging the interfering material.
  • the analysis device 20 By detecting the analyte by the analysis device 20 without the above process, the amplification and separation of the analyte and the interfering material and the detection of the analyte may be continuously performed. Accordingly, it is possible to reduce the time required for analysis by unifying the analysis steps, and to increase the convenience of analysis.

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  • Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)

Abstract

L'invention concerne un système d'analyse. La présente invention concerne en particulier un système d'analyse comprenant : un appareil de séparation destiné à agiter et séparer un échantillon contenant un analyte, une substance interférente et des particules magnétiques se liant à la substance interférente ; et un appareil d'analyse relié à l'appareil de séparation et irradiant l'échantillon avec de la lumière afin de détecter l'analyte au moyen d'un signal optique émis par celui-ci. L'appareil de séparation comprend : un corps ; une partie de réception disposée amovible sur le corps et destinée à recevoir l'échantillon ; une première partie électroaimant disposée sur le corps et destinée à générer un champ magnétique ; et une deuxième partie électroaimant disposée sur le corps pour faire face à la première partie électroaimant avec la partie de réception au centre, et destinée à générer un champ magnétique dans une direction différente de celle de la première partie électroaimant, au moins la première ou la deuxième partie électroaimant appliquant un champ magnétique sur les particules magnétiques, de sorte que l'analyte, la substance interférente et au moins certaines des particules magnétiques soient agités.
PCT/KR2021/004085 2020-04-03 2021-04-01 Système et procédé d'analyse WO2021201635A1 (fr)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2011158334A (ja) * 2010-01-29 2011-08-18 Beckman Coulter Inc 分析方法、分析装置および分析プログラム
KR101195957B1 (ko) * 2009-03-26 2012-10-30 한양대학교 에리카산학협력단 표면증강 라만 산란 복합 프로브 및 이를 이용하여 표적 물질을 검출하는 방법
KR101907267B1 (ko) * 2016-10-10 2018-10-11 서울대학교산학협력단 다공성 격벽 및 자석을 이용한 표적물질 검출용 키트
KR20190016066A (ko) * 2019-02-08 2019-02-15 서울대학교산학협력단 이미지 센서 및 자성체를 포함하는 표적물질 검출용 키트
KR20200034725A (ko) * 2017-07-19 2020-03-31 암젠 인크 자기 보조 분리 장치 및 관련 방법

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101195957B1 (ko) * 2009-03-26 2012-10-30 한양대학교 에리카산학협력단 표면증강 라만 산란 복합 프로브 및 이를 이용하여 표적 물질을 검출하는 방법
JP2011158334A (ja) * 2010-01-29 2011-08-18 Beckman Coulter Inc 分析方法、分析装置および分析プログラム
KR101907267B1 (ko) * 2016-10-10 2018-10-11 서울대학교산학협력단 다공성 격벽 및 자석을 이용한 표적물질 검출용 키트
KR20200034725A (ko) * 2017-07-19 2020-03-31 암젠 인크 자기 보조 분리 장치 및 관련 방법
KR20190016066A (ko) * 2019-02-08 2019-02-15 서울대학교산학협력단 이미지 센서 및 자성체를 포함하는 표적물질 검출용 키트

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