Methods and devices for analyte collection, extraction, concentration, and detection for clinical applications
In various embodiments devices and methods for the detection and/or quantification of clinically relevant pathogens (e.g., bacteria, fungi, viruses, etc.) are provided. In certain embodiments the device comprises a lateral-flow assay that detects the bacterium at a concentration of less than about 6×10 6 cells/mL, less than about 3×10 6 cells/ml, less than about 1×10 6 CFU/mL, or less than about 50 μg/mL. In certain embodiments the device comprises an aqueous two-phase system (ATPS) comprising a mixed phase solution that separates into a first phase solution and a second phase solution; and a lateral-flow assay (LFA). In certain embodiments the device comprises a flow-through system comprising a concentration component comprising an aqueous two-phase system (ATPS) comprising a mixed phase solution that separates into a first phase solution and a second phase solution; and a detection component disposed beneath said concentration component.
1. A wick that concentrates an analyte for application to an immunoassay, wherein:
said wick consists of five fiberglass sheets that form an elongate porous matrix, where said wick, in use, defines an upstream end and a downstream end, where the downstream end of said porous matrix tapers to a narrow region, and where said wick, upstream of the tapered region, comprises a dehydrated salt and a dehydrated polymer that when rehydrated form a mixed phase of an aqueous two-phase system, wherein:
said dehydrated salt is disposed in said wick upstream from said dehydrated polymer;
said polymer comprises polyethylene glycol; and
said wick does not contain a detection region.
2. The wick of claim 1 , wherein said salt comprises a salt selected from the group consisting of potassium phosphate, sodium sulfate, magnesium sulfate, ammonium sulfate, or sodium citrate.
3. The wick of claim 1 , wherein said salt comprises potassium phosphate.
4. A system for the detection and/or quantification of a bacterium, a fungus, or a virus in a sample, said system comprising:
a wick according to claim 1 ; and
a detection component disposed at the tip of said tapered region, wherein said detection component comprises a lateral flow assay.
5. The system of claim 4 , wherein:
said lateral flow assay comprises a conjugate pad, a reaction pad, and optionally a sink.
6. The system of claim 4 , wherein said lateral flow assay is configured for the detection of a bacterium.
7. The system of claim 6 , wherein said bacterium is an oral bacterium, a bacterium found in urine, a bacterium found in vaginal fluid, or a bacterium found on a vaginal swab, or a bacterium found on an endocervical swab.
8. The system of claim 7 , wherein:
said bacterium is an oral bacterium that comprises Prevotella sp., Porphyromonas sp., Streptococcus sp., Actinomyces viscosus, Lactobacillus casei, Staphylococcus aureus, Candida albicans, Lactobacillus acidophilus, Capnocytophaga gingivalis, Fusobacterium nucleatum , or Bacteriodes fortsythus ; or
said bacterium is a bacterium found in vaginal fluid where said bacterium comprises Trichomonas sp., Actinomyces sp., Gardnerella sp., Neisseria sp., Chlamydia sp., or Treponema sp; or
said bacterium is a bacterium found in urine where said bacterium comprises E. coli, Proteus sp., Trichomonas sp., Actinomyces sp., Gardnerella sp., Neisseria sp., Chlamydia sp., or Treponema sp.
9. A kit for the detection and/or quantification of a bacterium, said kit comprising:
a wick of claim 1 ; and
a collection device for collecting a biological sample.
10. The system of claim 4 , wherein said salt comprises a salt selected from the group consisting of potassium phosphate, sodium sulfate, magnesium sulfate, ammonium sulfate, and sodium citrate.
11. The system of claim 4 , wherein said salt comprises potassium phosphate.