Methods for isolating a target analyte from a heterogenous sample

• US 9,970,931 B2
• Filed: 10/24/2016
• Issued: 05/15/2018
• Est. Priority Date: 04/21/2010
• Status: Active Grant

• Current Assignee: DNAE Group Holdings Limited (Genting Bhd.)
• Original Assignee: DNAE Group Holdings Limited (Genting Bhd.)
• Inventors: Dryga, Sergey A., Esch, Victor C., Clarizia, Lisa-Jo Ann, Adams, Eddie W., Ung, Thearith H., Sitdikov, Ravil A.
• Primary Examiner(s): Navarro, Albert M
• Assistant Examiner(s): NAVARRO, ALBERT MARK
• Application Number:
  US15/332,679
• Publication Number:
  US 20170038374A1
• Time in Patent Office: 568 Days
• CPC Class Codes: C07K 16/1203 from Gram-negative bacteria
• CPC Class Codes: C07K 16/1267 from Gram-positive bacteria
• CPC Class Codes: C07K 2317/33 Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
• CPC Class Codes: C12N 13/00 Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
• CPC Class Codes: C12Q 1/24 Methods of sampling, or inoculating or spreading a sample; Methods of physically isolating an intact microorganisms
• CPC Class Codes: G01N 1/34 Purifying; Cleaning processes or apparatus for extracting or separating nucleic acids from biological samples C12N15/1003
• CPC Class Codes: G01N 33/54333 Modification of conditions of immunological binding reaction, e.g. use of more than one type of particle, use of chemical agents to improve binding, choice of incubation time or application of magnetic field during binding reaction
• CPC Class Codes: G01N 33/569 for microorganisms, e.g. protozoa, bacteria, viruses

Abstract

The invention generally relates to methods of using compositions that include sets of magnetic particles, members of each set being conjugated to an antibody specific for a pathogen, and magnets to isolate a pathogen from a body fluid sample.

Claims

1. A method for isolating at least one pathogen from a heterogeneous sample, the method comprising:

obtaining a heterogeneous sample comprising at least one pathogen;

incubating the sample with a composition comprising a plurality of sets of magnetic particles specific to one or more pathogens, wherein members of a first set of magnetic particles are conjugated to a first class of capture moiety, and members of a second set of magnetic particles are conjugated to a second class of capture moiety; and

separating particle bound pathogen from other components in the sample;

wherein the first and second classes of capture moieties are selected from the group consisting of antibodies, lectins, bacteriophages, antimicrobial agents, and oligonucleotides; and

wherein the first and second classes are different.

2. The method of claim 1, wherein each set of magnetic particles is present at a concentration designed for detection of a specific pathogen in said sample.

3. The method of claim 2, wherein the first and second set of magnetic particles are provided at different concentrations.

4. The method of claim 1, wherein the pathogen is selected from the group consisting of fungi, bacteria and a combination thereof.

5. The method of claim 4, wherein the fungi comprises two or more species of fungi selected from the group consisting of the Candida genus, Aspergillus genus, and Cryptococcus genus.

6. The method of claim 4, wherein the bacteria comprise one or species of bacteria selected from the group consisting of Staphylococcus genus, Enterobacteriaceae genus, Acinetobacter genus, Strenotrophomonas genus, Pseudomonas genus, Neisseria genus, Clostridium genus, and Enterococcus genus.

7. The method of claim 1, wherein members of the first class are different.

8. The method of claim 1, wherein members of the first class are the same.

9. The method of claim 1, wherein members of the second class are different.

10. The method of claim 1, wherein members of the second class are the same.

11. The method of claim 1, further comprising identifying the pathogen.

12. The method of claim 1, further comprising lysing the separated pathogen to release nucleic acid.

13. The method of claim 11, further comprising conducting an assay on the nucleic acid to identify the pathogen.

14. The method of claim 13, wherein the assay is selected from the group consisting of a sequencing reaction and a polymerase chain reaction.

15. The method of claim 13, wherein the assay is a multiplex assay.