Apparatus and method for point-of-collection measurement of biomolecular reaction
A technology of biomolecules and reactions, used in branch equipment, biological testing, chemical instruments and methods, etc.
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Embodiment I
[0119] Example I: Quantitative Measurement of 25-Hydroxyvitamin D
[0120] In this example, using the Figure 7 A modular diagnostic testing platform ("test strip") 704 is shown. Included on the test strip 704 is a blood filter assembly 717 for filtering the raw blood sample into a plasma sample. The filter assembly 717 consists of a stack of reduced pore size commercial filters (Whatman, Kent, UK) ranging in pore size from approximately 2.7 μm (considering the approximate limit for filtering highly deformed red blood cells) to 0.7 μm (small enough to filter platelets). (It is worth noting that the pore size of these filters was not optimal for reproducibility, accuracy, and speed in this work).
[0121] The circulating concentration of a metabolite of vitamin D, 25-hydroxyvitamin D [25(OH)D], is considered the best indicator of vitamin D status. We have developed a nanoparticle-based assay and test strips that quantify 25(OH)D concentrations between 5 ng / ml and 75 ng / ml, ...
Embodiment II
[0126] Example II: Detection of Kaposi's Sarcoma-Associated Herpesvirus (KSHV) DNA
[0127] In this example, using image 3 A modular diagnostic test platform shown as 203, as described above.
[0128] BLAST primer design was used to design short DNA sequences for use as probes for KHSV DNA. Briefly, oligonucleotides were selected that were specific for KSHV DNA encoding vCyclin. Gold nanoparticles are attached to oligonucleotides via a 5' alkylthiol group. The nanoparticles have an average diameter of 15 nm, which is a compromise between the higher sensitivity of larger particles and the easier handling of the properties and stability of smaller particles. Add 100 μL of 100 μM KSHV to every 1 ml of 3 nM GNP solution, and react overnight. Then, concentrated solutions of sodium phosphate and sodium dodecyl sulfate (SDS) were added to make them respectively reach concentrations of 10 mM and 0.01% in the solution, and reacted overnight again. Next, sodium chloride was added ...
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