Pathogen detection
Pathogens are detected through the use of mutation-resistant ligands.
1. An article comprising:
a substrate; and
one or more mutation-resistant ligands affixed in a predetermined periodic pattern to at least a portion of the substrate, wherein the one or more mutation-resistant ligands is affixed to the substrate via a linker, wherein the one or more mutation-resistant ligands can be bound by pathogens wherein the predetermined periodic pattern is indicative of the pathogen to be selectively bound to the mutation resistant ligand and is recognizable when a pathogen is bound to the ligand, and
wherein the mutation-resistant ligands are involved in functions important to or essential to virulence of the pathogens.
2. The article of claim 1 wherein the linker comprises bovine serum albumin or polyethylene glycol.
3. The article of claim 1 wherein the mutation-resistant ligand comprises a siderophore, a glycan, or a neuraminidase inhibitor.
4. The article of claim 3 wherein the glycan comprises a cell surface molecule involved in a pathogen infecting a host cell.
5. The article of claim 3 wherein the neuraminidase inhibitor comprises oseltamivir or a derivative thereof.
6. The article of claim 1 wherein the one or more mutation-resistant ligands comprises a first mutation-resistant ligand and a second mutation-resistant ligand.
7. The article of claim 6 wherein the first mutation-resistant ligand selectively binds a first pathogen and the second mutation-resistant ligand selectively binds to a second pathogen different from said first pathogen.
8. The article of claim 6 wherein the first mutation-resistant ligand is affixed to the substrate in a first predetermined pattern and the second mutation-resistant ligand is affixed to the substrate in a second predetermined pattern.
9. A method comprising:
contacting a biological sample comprising a pathogen with the article of claim 1 for a time sufficient to allow the one or more mutation-resistant ligands to selectively bind to the pathogen; and
detecting the pathogen bound to the one or more mutation-resistant ligands.
10. The method of claim 9 wherein the at least one mutation-resistant ligand comprises a first mutation-resistant ligand that selectively binds to a first pathogen and a second mutation-resistant ligand that selectively binds to a second pathogen.
11. The method of claim 9 wherein the at least one mutation-resistant ligand comprises a first mutation-resistant ligand and a second mutation-resistant ligand, each of which selectively binds the same pathogen.
12. The method of claim 9 wherein detecting the pathogen bound to the one or more mutation-resistant ligands comprises labeling the bound pathogen and detecting the label.
13. The method of claim 9 wherein detecting the pathogen bound to the one or more mutation-resistant ligands comprises label-free detection.
14. The method of claim 13 wherein label-free detection comprises optical imaging.
15. The method of claim 14 wherein the label-free detection further comprises recognizing the predetermined pattern produced by the pathogen selectively bound to the one or more mutation-resistant ligands.
16. The method of claim 15 wherein:
the one or more mutation-resistant ligand comprises:
a first mutation-resistant ligand that selectively binds to a first pathogen and is affixed to the substrate in a first predetermined pattern; and
a second mutation-resistant ligand that selectively binds to a second pathogen and is affixed to the substrate in a second predetermined pattern; and
the label-free detection comprises recognizing the first predetermined pattern and the second predetermined pattern.
17. The method of claim 15 wherein recognizing the predetermined pattern comprises using a two-dimensional fast Fourier transform.
18. The method of claim 9 wherein the pathogen comprises a bacterium, a virus, a parasite, a protozoan, a protist, or a fungus.
19. A device comprising:
an analytical chamber;
a sample reservoir in fluid communication with the analytical chamber;
the article of claim 1 ; and
an image recorder.
20. The device of claim 19 further comprising:
a dielectrophoretic concentrator in fluid communication between the analytical chamber and the sample reservoir; and
a wash reservoir in fluid communication with the dielectrophoretic concentrator.
21. The device of claim 20 further comprising:
a pump in functional communication with the sample reservoir and/or a pump in functional communication with the wash reservoir; and
a control processing unit in functional communication with at least one of the pumps.
22. The device of claim 19 further comprising at least one of:
a magnifying lens functionally positioned between the article and the image recorder;
an illumination source situated to illuminate the article;
a control processing unit in functional communication with the image recorder; and
a user interface display functionally connected to the image recorder.