Target capture system

• US 9,599,610 B2
• Filed: 12/16/2013
• Issued: 03/21/2017
• Est. Priority Date: 12/19/2012
• Status: Active Grant

• Current Assignee: DNAE Group Holdings Limited (Genting Bhd.)
• Original Assignee: DNAE Group Holdings Limited (Genting Bhd.)
• Inventors: Sitdikov, Ravil A., Adams, Eddie W., Torrance, Magdalena A., Aley, David K., Smith, Erik J., Esch, Victor C.
• Primary Examiner(s): Nickol, Gary
• Assistant Examiner(s): JACKSON-TONGUE, LAKIA J
• Application Number:
  US14/107,315
• Publication Number:
  US 20140170652A1
• Time in Patent Office: 1,191 Days
• US Class Current: 1/1
• CPC Class Codes: A23C 9/144 by electrical means, e.g. electrodialysis
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: C12N 1/20 Bacteria; Culture media therefor
• CPC Class Codes: C12N 15/1013 by using magnetic beads
• CPC Class Codes: C12Q 1/6827 for detection of mutation or polymorphism
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 1/689 for bacteria
• CPC Class Codes: C12Q 1/6895 for plants, fungi or algae
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: C12Q 2600/16 Primer sets for multiplex assays
• CPC Class Codes: G01N 2469/00 Immunoassays for the detection of microorganisms
• CPC Class Codes: G01N 33/54326 Magnetic particles

Abstract

The invention generally relates to a system for isolating or separating a target from a sample. In certain aspects, processes performed by the target capture system include introducing a plurality of magnetic particles, in which a plurality of the particles include at least one binding moiety specific to a target, into a sample to form at least one target/particle complex and applying a magnetic field to isolate the magnetic particle/target complexes from the sample. The process starts at inputting a sample into the system and ends at delivering a capture target or nucleic acids of the target into a container for further analysis.

Claims

1. A method for isolating a pathogen from a sample, the method comprising

removably coupling a vessel comprising a pathogen-containing sample to a cartridge via a cartridge interface that is configured to provide communication between the vessel and the cartridge;

flowing magnetic particles conjugated to a capture moiety from the cartridge into the vessel to form a mixture;

flowing the mixture from the vessel into the cartridge;

flowing a fluid from the cartridge into the vessel to flush residual mixture from the vessel into the cartridge; and

incubating the mixture in the cartridge to allow the particles to bind to the pathogen;

applying a magnetic field to capture pathogen/magnetic particle complexes on a surface of a first magnetic trap;

washing the captured pathogen/magnetic particle complexes with a wash solution, thereby isolating the pathogen/magnetic particle complexes;

removing the magnetic field;

re-introducing the wash solution, thereby re-suspending the pathogen/magnetic particle complexes;

flowing the re-suspended pathogen/magnetic particle complexes through a channel and into a second magnetic trap; and

engaging the second magnetic trap, thereby re-capturing the pathogen/magnetic particle complexes;

wherein the first magnetic trap is configured to capture the pathogen/magnetic particle complexes from a macro-scale volume of fluid, and the second magnetic trap is configured to concentrate substantially all of the pathogen/magnetic particle complexes into a micro-scale volume of fluid.

2. The method of claim 1, further comprising lysing the captured pathogen in a region selected from the group consisting of the first magnetic trap, the second magnetic trap, and a combination thereof.

3. The method of claim 2, further comprising separating nucleic acid from the lysate;

and collecting the separated nucleic acid into a vial.

4. The method of claim 3, further comprising analyzing the nucleic acid to thereby identify the pathogen.

5. The method of claim 4, wherein analyzing comprises conducting a sequencing reaction.

6. The method of claim 4, wherein analyzing comprises conducting an amplification reaction.

7. The method of claim 6, wherein the vial comprises at least one detectably labeled nucleic acid probe specific for the pathogen.

8. The method of claim 1, wherein the sample comprises a plurality of pathogen.

9. The method of claim 8, wherein the plurality of magnetic particles are composed of different sets, wherein members of the different sets are conjugated to different capture moieties that are specific for the different pathogen.

10. The method of claim 9, further comprising conducting a multiplex reaction to analyze nucleic acid from the plurality of isolated pathogen and thereby identify the plurality of pathogen.

11. The method of claim 10, wherein the multiplex reaction is a multiplex sequencing reaction.

12. The method of claim 10, wherein the multiplex reaction is a multiplex amplification reaction.

13. The method of claim 1, wherein the capture moiety is a type selected from the group consisting of antibodies, lectins, bacteriophages, antimicrobial agents, oligonucleotides, and combinations thereof.

14. The method of claim 1, wherein the pathogen is selected from the group consisting of fungi, bacteria or both.

15. The method of claim 14, wherein the fungi are selected from the group consisting of the Candida genus, the Aspergillus genus, the Cryptococcus genus, and a combination thereof.

16. The method of claim 14, wherein the bacteria are selected from the group consisting of the Staphylococcus genus, the Enterobacteriaceae genus, the Acinetobacter genus, the Stenotrophomas genus, the Pseudomonas genus, the Neisseria genus, the Clostridium genus, and the Enterococcus genus.

17. The method of claim 1, wherein said sample is a human tissue or body fluid.