Diagnostic and sample preparation devices and methods
Inventors
Kelley, Shana O. • Bortolin, Susan • Orton, Reginald James McKenzie • Wiechula, Stefan Christopher
Assignees
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Publication Number
US-9580742-B2
Publication Date
2017-02-28
Expiration Date
Abstract
Contemplated methods and devices are drawn to preparation and analysis of analytes from biological samples. In a preferred embodiment the analytes are nucleic acids that are both released from biological compartment present in the sample and fragmented through the use of a voltage potential applied to a pair of electrodes. The nucleic acids thus prepared are subsequently characterized.
Core Innovation
The invention relates to analyzing a biological sample using an extraction zone that comprises a pair of electrodes. A first electrical signal is applied to the pair of electrodes so that a plurality of nucleic acids are released from the biological sample and fragmented within the extraction zone to produce a plurality of fragmented nucleic acids.
Selecting the first electrical signal comprises selecting at least one of a magnitude of a voltage applied to the pair of electrodes, a duration of a voltage pulse applied to the pair of electrodes, and a frequency at which a voltage pulse is applied to the pair of electrodes. Fragmentation is controlled based on a desired average length in bases of the plurality of fragmented nucleic acids, with reduced fragment lengths and average lengths at or below specified thresholds in bases.
The invention states that controlling nucleic acid fragmentation reduces subsequent analysis time for hybridization and supports direct electrochemical detection. The analysis architecture includes electrodes for sensing and reference in a hybridization-responsive reporting context using probes complementary to fragmented nucleic acids, and nanostructured sensing electrodes with a spiky, rough, or fractal surface are described as part of the electrochemical detection approach.
Claims Coverage
The independent claim defines a method of analyzing a biological sample by releasing and fragmenting nucleic acids in an extraction zone using a pair of electrodes, where fragmentation is tuned by selecting electrical signal parameters based on a desired average fragment length. The dependent claims add fragment-length constraints, a hybridization-time improvement threshold, enhancing reagent types, and specific sensing-electrode structure.
Electrode-based release and fragmentation in an extraction zone
A method of analyzing a biological sample by transferring the biological sample to an extraction zone comprising a pair of electrodes, and applying a selected first electrical signal to release a plurality of nucleic acids and fragment them within the extraction zone to produce a plurality of fragmented nucleic acids.
Selecting electrical signal parameters based on desired average length
Selecting the first electrical signal comprises selecting, based on a desired average length in bases of the plurality of fragmented nucleic acids, at least one of a magnitude of a voltage applied to the pair of electrodes, a duration of a voltage pulse applied to the pair of electrodes, and a frequency at which a voltage pulse is applied to the pair of electrodes.
Fragment-length constraints for the produced fragments
The plurality of fragmented nucleic acids have average lengths that meet specified base-length limits relative to a plurality of nucleic acids prior to fragmentation and/or to an absolute base-length threshold.
Reduced hybridization time due to fragmentation
The plurality of nucleic acids are fragmented so they can hybridize to a solid-phase bound capture probe in a hybridization time at least 25% shorter than the hybridization time before fragmentation.
Hybridization improvement with enhancing reagents
An enhancing reagent is chosen from metal ion, free radical promoting compound, chaotropic salt, ionic detergent, and nonionic detergent.
Nanostructured microelectrode sensing electrode
A sensing electrode is a nanostructured microelectrode with a spiky, rough, or fractal surface.
Overall, the claim coverage centers on applying a selected electrical signal to a pair of electrodes in an extraction zone to release and fragment nucleic acids, where the electrical signal is chosen to achieve a desired average fragment length. Dependent claim coverage adds fragment-length thresholds, a quantified hybridization-time reduction, enhancing-reagent categories, and a defined nanostructured sensing-electrode surface morphology.
Stated Advantages
Reduced hybridization time for hybridization to a solid-phase bound capture probe, at least 25% shorter than hybridization time before fragmentation.
Documented Applications
Analyzing a biological sample using an extraction zone and a pair of electrodes to release and fragment nucleic acids for downstream hybridization and electrochemical analysis.
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