Methods for isolating a target analyte from a heterogeneous sample

• US 9,476,812 B2
• Filed: 07/26/2013
• Issued: 10/25/2016
• Est. Priority Date: 04/21/2010
• Status: Active Grant

• Current Assignee: DNAE Group Holdings Limited (Genting Bhd.)
• Original Assignee: DNA Electronics, Inc. (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 MARK
• Application Number:
  US13/952,130
• Publication Number:
  US 20130316355A1
• Time in Patent Office: 1,187 Days
• US Class Current: 1/1
• 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 pathogens from a heterogeneous sample, the method comprising

obtaining a heterogeneous sample comprising pathogens;

introducing the heterogeneous sample into a fluidic device through a vessel interface of the fluidic device;

introducing to the heterogeneous sample from a reagent storage chamber of the fluidic device a composition comprising at least one magnetic particle conjugated to at least two sets of capture moieties, wherein each set of capture moieties is specific for different pathogen,

incubating in the fluidic device the sample and the composition; and

identifying the different pathogens in the sample.

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

3. The method of claim 1, wherein the at least two sets are provided at different concentrations.

4. The method of claim 1, wherein the capture moieties are selected from the group consisting of antibodies, lectins, bacteriophages, antimicrobial agents, and oligonucleotides.

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

6. The method of claim 5, further comprising conducting an assay on the nucleic acid to identify the pathogen.

7. The method of claim 6, wherein the assay is selected from the group consisting of a sequencing reaction and a polymerase chain reaction.

8. The method of claim 7, wherein the assay is a multiplex assay.