Target detection

• US 10,379,113 B2
• Filed: 01/23/2017
• Issued: 08/13/2019
• 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: Norvell, Meghan E.
• Primary Examiner(s): Bhat, Narayan K
• Application Number:
  US15/413,008
• Publication Number:
  US 20170131270A1
• Time in Patent Office: 932 Days
• CPC Class Codes: G01N 33/54326 Magnetic particles
• 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 for indicating whether an assay for isolating targets is properly isolating and detecting targets. Methods of the invention involve obtaining a sample suspected of a containing target, introducing a detectable marker into the sample, conducting an assay using magnet particles to isolate the detectable marker and the target if it is present in the sample, determining the presence or absence of the target; and confirming that the assay functioned properly by determining presence or absence of the detectable marker.

Claims

1. A method for detecting a target and confirming proper functioning of an assay, the method comprising

obtaining a sample suspected of containing a target;

introducing a detectable marker into the sample;

incubating the sample to allow for coupling of the detectable marker and the target, if present, to magnetic particles comprising both a first binding moiety specific to the target and a second binding moiety specific to the detectable marker in the sample;

conducting an assay using magnetic particles to isolate the detectable marker and the target from the sample;

determining the presence or absence of the target in the sample; and

confirming proper function of the assay by determining presence or absence of the detectable marker.

2. The method of claim 1, wherein the detectable marker comprises a microbe.

3. The method of claim 2, wherein the microbe is detectable via a microbe-sensitive chromogen.

4. The method of claim 1, wherein the detectable marker comprises a label.

5. The method of claim 4, wherein the label is a fluorescent label.

6. The method of claim 1, wherein the target is selected from the group consisting of cells, bacteria, viruses, fungi, and any combination thereof.

7. The method of claim 1, wherein the assay comprises exposing the magnetic particles to a magnetic field.

8. The method of claim 7, wherein the assay further comprises

flowing the sample through a channel comprising surface binding moieties specific to the targets and to the detectable marker attached to a surface of the channel;

and binding the magnetic particles to the surface of the channel, wherein the step of applying a magnetic field brings the magnetic particles coupled to the target and detectable marker into proximity of the surface to facilitate binding of magnetic particles to the surface of the channel; and

washing away unbound magnetic particles and unbound sample components.

9. The method of claim 1, wherein the method further comprises introducing the magnetic particles into the sample, wherein the magnetic particles are introduced prior to the conducting an assay step.

10. The method of claim 1, wherein the presence of the target is determined separately from the presence of the detectable marker.