Target detection
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.
1. A method for detecting a pathogen and confirming proper functioning of an assay, the method comprising
obtaining a sample suspected of containing a pathogen;
introducing a detectable marker into the sample;
introducing magnetic particles into the sample;
incubating the sample to allow for coupling of the detectable marker and the pathogen to the magnetic particles, wherein the detectable marker and the pathogen couple to separate binding moieties;
conducting an assay using the magnetic particles to isolate and determine presence of both the detectable marker from the sample and the pathogen, wherein the presence of the pathogen is determined separate from the presence of the detectable marker, and wherein determination of the presence of the detectable marker confirms proper functioning of the assay.
2. The method of claim 1 , wherein the magnetic particles are divided into a first set and a second set.
3. The method of claim 2 , wherein the first and second sets of magnetic particles are labeled.
4. The method of claim 3 , wherein, the labels are different between the first set and second set.
5. The method of claim 2 , wherein members of the first set of magnetic particles are coupled to a binding moiety specific to the pathogen to form magnetic particle/pathogen complexes, and members of the second set of magnetic particles are coupled to a binding moiety specific to the detectable marker to form a magnetic particle/detectable marker complex.
6. The method of claim 5 , wherein the assay comprises exposing the magnetic particle/pathogen complexes and the magnetic particle/detectable marker complexes to a magnetic field.
7. The method of claim 6 , wherein the assay further comprises flowing the sample through a channel comprising surface binding moieties specific to the pathogen and to the detectable marker attached to a surface of the channel; and
binding the pathogen/magnetic particle complexes and detectable marker/magnetic particle complex to the surface of the channel, wherein the step of applying a magnetic field brings the pathogen/magnetic particle complexes and detectable marker/magnetic particle complexes into proximity of the surface to facilitate binding of the pathogen/magnetic particle complexes and the detectable marker/magnetic particle complex to the surface of the channel; and
washing away unbound magnetic particles and unbound sample components.
8. The method of claim 1 , wherein the detectable marker comprises a microbe.
9. The method of claim 8 , wherein the microbe is detectable via a microbe-sensitive chromogen.
10. The method of claim 1 , wherein a plurality of the magnetic particles comprise two or more different binding moieties.
11. The method of claim 10 , wherein a first binding moiety is specific to the pathogen, and a second binding moiety is specific to the detectable marker.
12. The method of claim 11 , wherein the assay comprises exposing the magnetic particles to a magnetic field.
13. The method of claim 12 , wherein the assay further comprises flowing the sample through a channel comprising surface binding moieties specific to the pathogen 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 containing the pathogen/magnetic particle complexes and detectable marker/magnetic particle complexes 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.