METHOD FOR PRETREATING SPECIMEN AND METHOD FOR ASSAYING BIOLOGICAL SUBSTANCE

• US 20110262919A1
• Filed: 12/25/2009
• Published: 10/27/2011
• Est. Priority Date: 12/25/2008
• Status: Active Grant

• Current Assignee: Universal Bio Research Co., Ltd (Precision System Science Co., Ltd)
• Original Assignee: Universal Bio Research Co., Ltd (Precision System Science Co., Ltd)
• Inventors: Tajima, Hideji
• Granted Patent: US 9,182,395 B2
• Time in Patent Office: Days
• US Class Current: 435/6.11
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 3/508 rigid containers not provided for above
• CPC Class Codes: B01L 3/52 Containers specially adapted for storing or dispensing a reagent B01L3/02 takes precedence; containers for medical or pharmaceutical purposes A61J1/00; containers in general B65D; storing or dispensing test elements G01N33/4875; automated reagent dispensing G01N35/1002
• CPC Class Codes: C12Q 1/6804 Nucleic acid analysis using immunogens immunoassay G01N33/53
• CPC Class Codes: C12Q 2563/143 Magnetism, e.g. magnetic label
• CPC Class Codes: G01N 33/54326 Magnetic particles
• CPC Class Codes: G01N 33/54386 Analytical elements
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
• CPC Class Codes: G01N 35/026 having blocks or racks of reaction cells or cuvettes
• CPC Class Codes: G01N 35/1009 Characterised by arrangements for controlling the aspiration or dispense of liquids
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]
• CPC Class Codes: Y10T 436/25125 Digestion or removing interfering materials

Abstract

On magnetic particles serving as a first support, an antibody against a nonspecific reaction factor is immobilized. These magnetic particles are mixed with a specimen and suspended therein. After suspending, the suspension is sucked up into a pipette chip and a magnet comes close to the pipette chip. While remotely constraining with the magnet the magnetic particles carrying the nonspecific reaction factor bonded thereto, the residual liquid is discharged into a well. Thus, the removal of a contaminant contained in the specimen is completed. The thus treated specimen discharged into the well is subjected to an immunoassay. The magnetic particles carrying the antibody immobilized thereon are mixed with the treated specimen and suspended therein. The suspension is sucked up into a pipette chip and the magnetic particles carrying an antigen bonded thereto are separated by use of the magnet. The magnetic particles carrying the antigen bonded thereto are washed, mixed with an enzyme-labeling solution, which contains a second support, and suspended therein. After suspending, the magnetic particles being labeled and carrying the antigen bonded thereto are mixed with a substrate solution and subjected to the measurement of emission intensity, etc.

Claims

1. A system for assaying a biologically-relevant substance in a specimen, comprising:

a first support to which a substance having affinity to a contaminant contained in the specimen, a substance that inactivates the contaminant, or a substance having affinity to the biologically-relevant substance in the specimen is immobilized; and

a second support selected from a support to which a reagent for detecting the biologically-relevant substance is immobilized and a support made of a solid-phased reagent for detecting the biologically-relevant substance.

2. The system according to claim 1, wherein the biologically-relevant substance is an antigen or an antibody.

3. The system according to claim 1, wherein the biologically-relevant substance is a nucleic acid.

4. The system according to claim 1, wherein the contaminant is a nonspecific reaction factor.

5. The system according to claim 4, wherein the nonspecific reaction factor is at least one selected from the group consisting of an immunoglobulin, an M protein, a heterophilic antibody and a rheumatoid factor.

6. The system according to claim 1, wherein the first support is at least one selected from the group consisting of magnetic particles, a gel, a resin and a membrane.

7. The system according to claim 6, wherein the magnetic particles are held in a dispensing chip.

8. The system according to claim 1, wherein the reagent for detecting the biologically-relevant substance comprises a labeled antigen or a labeled antibody against the biologically-relevant substance or a primer and a probe for amplifying the biologically-relevant substance.

9. The system according to claim 1, wherein solid-phasing is carried out by freeze-drying.

10. The system according to claim 1, wherein the specimen, the first support and the second support are respectively held in different holding portions.

11. The system according to claim 10, wherein:

a specimen holding portion in which the specimen is held;

a first holding portion in which the first support is held; and

a second holding portion in which the second support is held

are arranged in an approximate straight line.

12. The system according to claim 10, wherein:

the specimen holding portion in which the specimen is held;

the first holding portion in which the first support is held; and

the second holding portion in which the second support is held

are integrated into a cartridge.

13. The system according to claim 10, wherein the holding portions are sealable.

33. The method according to claim 20, wherein the pretreatment of the specimen and the assay of the treated specimen are carried out using the system according to claim 1.

34. The method according to claim 33, wherein the pretreatment of the specimen and the assay of the treated specimen are carried out successively.

14. A cartridge comprising:

a first holding portion, in which a first support to which a substance having affinity to a contaminant contained in a specimen, a substance that inactivates the contaminant, or a substance having affinity to the biologically-relevant substance in the specimen is immobilized is held in advance;

a second holding portion, in which a second support selected from a support to which a reagent for detecting the biologically-relevant substance is immobilized and a support made of a solid-phased reagent for detecting the biologically-relevant substance is held; and

a specimen holding portion in which the specimen is held.

15. The cartridge according to claim 14, wherein the first support is magnetic particles.

16. The cartridge according to claim 15, wherein the magnetic particles are held in a dispensing chip.

17. The cartridge according to claim 14, wherein solid-phasing is carried out by freeze-drying.

18. The cartridge according to claim 14, wherein the holding portions are sealable.

19. The cartridge according to claim 14, wherein:

the specimen holding portion in which the specimen is held;

the first holding portion in which the first support is held; and

the second holding portion in which the second support is held

are arranged in an approximate straight line.

30. A method for assaying a biologically-relevant substance, wherein a specimen pretreated using the method according to claim 18 is provided to an assay device to assay the biologically-relevant substance in the specimen.

31. The method according to claim 30, wherein the assay is an immunoassay or an assay by means of a nucleic acid amplification method.

32. The method according to claim 31, wherein the nucleic acid amplification method is a PCR method.

20. A method for pretreating a specimen before assaying a biologically-relevant substance in the specimen, comprising:

a step of treating the specimen using a first support to which a substance having affinity to a contaminant contained in the specimen, a substance that inactivates the contaminant, or a substance having affinity to the biologically-relevant substance in the specimen is immobilized.

21. The method according to claim 20, further comprising a step of preparing a second support selected from a support to which a reagent for detecting the biologically-relevant substance is immobilized and a support made of a solid-phased reagent for detecting the biologically-relevant substance.

22. The method according to claim 20, wherein the biologically-relevant substance is an antigen or an antibody.

23. The method according to claim 20, wherein the biologically-relevant substance is a nucleic acid.

24. The method according to claim 20, wherein the contaminant is a nonspecific reaction factor.

25. The method according to claim 24, wherein the nonspecific reaction factor is at least one selected from the group consisting of an immunoglobulin, an M protein, a heterophilic antibody and a rheumatoid factor.

26. The method according to claim 20, wherein the first support is at least one selected from the group consisting of magnetic particles, a gel, a resin and a membrane.

27. The method according claim 26, wherein the magnetic particles are treated in a dispensing chip.

28. The method according to claim 21, wherein the reagent for detecting the biologically-relevant substance comprises a labeled antigen or a labeled antibody against the biologically-relevant substance or a primer and a probe for amplifying the biologically-relevant substance.

29. The method according to claim 21, wherein solid-phasing is carried out by freeze-drying.

35. A nucleic acid assay device, comprising:

(a) a specimen holding portion in which a specimen is held;

(b) a first holding portion in which trapping particles for trapping a target nucleic acid from the specimen are held;

(c) a second holding portion in which a reagent for detecting the target nucleic acid is held;

(d) a dispensing mechanism for dispensing the specimen into the specimen holding portion, a mechanism for mixing the specimen with the trapping particles to extract the target nucleic acid from the specimen, and a mechanism for mixing the extracted target nucleic acid with the reagent for detecting; and

(e) a mechanism selected from the group consisting of;

a mechanism for pouring a hydrophobic fluid, which has a specific gravity smaller than that of a mixed fluid of the target nucleic acid and the reagent for detecting, into the second holding portion;

a mechanism for removing or putting lids for covering the respective holding portions;

a mechanism for irradiating an irradiating light for letting the target nucleic acid fluoresce;

a mechanism for receiving a light from the target nucleic acid irradiated with the irradiating light to detect the target nucleic acid; and

a mechanism in which the mechanisms are combined.

36. The nucleic acid assay device according to claim 35, wherein the trapping particles are magnetic particles.

37. The nucleic acid assay device according to claim 35, wherein the magnetic particles are held in a dispensing chip.

38. The device according to claim 35, wherein the hydrophobic fluid is a mineral oil having 16 to 20 carbon atoms.

39. The device according to claim 35, wherein the aforementioned (d) and the mechanisms in (d) have a mechanism for sliding upper portions of the holding portions.

40. The device according to claim 35, wherein the mechanisms in (d) and the mechanism in (e) or a mechanism in which the mechanisms in (d) and the mechanism in (e) are combined are installed in the same nozzle.

41. The device according to claim 35, wherein the specimen holding portion, the first holding portion and the second holding portion are arranged in an approximate straight line.

42. The device according to claim 35, wherein the reagent for detecting comprises a reagent for nucleic acid amplification using a PCR method or an isothermal amplification method and a reagent for detecting an amplified product.

43. The device according to claim 35, wherein the reagent for detecting is freeze-dried in advance.

44. The device according to claim 35, wherein the first and second holding portions are formed into a cartridge.

45. A method for assaying a nucleic acid, wherein the nucleic acid is extracted and detected using the device according to claim 35.