Apparatus and method for fast digital detection
Inventors
Handique, Kalyan • Lebofsky, Ronald • Shinoff, Josh
Assignees
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Publication Number
US-12290813-B2
Publication Date
2025-05-06
Expiration Date
Abstract
Methods and systems for sample target molecules are provided.
Core Innovation
The disclosed invention is a method of detecting a target nucleic acid in multiple samples by contacting different samples comprising sample nucleic acids, optionally contained in vehicles, with different pluralities of solid supports to form different mixtures. The solid supports comprise a plurality of capture molecules that bind to the sample target nucleic acids or to vehicles comprising the target nucleic acid, and the solid supports contacted to different samples have a characteristic distinguishable from other solid supports contacted to other samples.
The method binds the capture molecules to the sample target nucleic acids or vehicles comprising the target nucleic acids, with a number of solid supports sufficiently high such that at least some solid supports are not bound to target nucleic acids or vehicles comprising target nucleic acids. The different mixtures are combined to form a bulk mixture, and the solid supports comprising bound sample target nucleic acids or vehicles are introduced into an array of wells.
Detection is performed with a scanner in each well to identify the presence or absence of the sample target nucleic acids and, in one variant, a signal and/or the characteristic of the solid support or the position of the wells. A computer processor determines the number of sample target nucleic acids per sample, wherein the determining comprises determining the number of wells having detected sample target nucleic acids, and the sample identity is determined by the detected characteristic of the solid support or by the detected position of the wells.
Claims Coverage
The partial content provides three independent claims, each covering digital detection across multiple samples using capture-molecule solid supports, combined loading into an array of wells, scanner-based detection, and computer-processor determination of target nucleic acid quantity per sample. The independent claims differ mainly in how sample identity is determined and whether detection uses presence or absence alone or presence or absence with signal intensity.
Distinguishable capture-solid supports for sample-specific mixtures
The method contacts different samples with different pluralities of solid supports that comprise capture molecules binding to the sample target nucleic acids or vehicles comprising the target nucleic acids, and the solid supports contacted to different samples have a characteristic distinguishable from other solid supports contacted to other samples.
Sufficient unbound solid supports to support digital well counting
The method binds the capture molecules to sample target nucleic acids or vehicles, wherein the number of solid supports is sufficiently high that at least some solid supports are not bound to target nucleic acids or vehicles comprising target nucleic acids.
Single-support well loading with scanner detection and processor-based quantification
The method combines the different mixtures to form a bulk mixture, introduces the solid supports into an array of wells where each well can contain one but no more than one solid support, detects in each well the presence or absence of the sample target nucleic acids and the characteristic of the solid support with a scanner, and determines with a computer processor the number of sample target nucleic acids per sample based on the number of wells having detected sample target nucleic acids, with sample identity determined by the detected characteristic.
Recorded well position linking wells to samples
The method introduces the solid supports into an array of wells where each well can contain no more than one solid support, records the position of wells receiving the sample beads such that wells corresponding to different samples is recorded, detects in each well the presence or absence of the sample target nucleic acids and the position of the wells with a scanner, and determines with a computer processor the number of sample target nucleic acids per sample based in part on the number of wells comprising detected sample target nucleic acids, where sample identity is determined by the position of the wells.
Multi-support wells with signal intensity and characteristic-based identity
The method introduces the solid supports into an array of wells where the wells can contain more than one solid support, detects in each well the presence and intensity or absence of signal indicative of the sample target nucleic acids and the characteristic of the solid support with a scanner, and determines with a computer processor the number of sample target nucleic acids per sample by determining the number of wells having detected sample target nucleic acids and different signal intensities together with presence or absence of signal in each well, where sample identity is determined by the detected characteristic.
Across the independent claims, the shared inventive core is digital nucleic-acid detection in multiple samples using capture-molecule solid supports, selecting a sufficiently high number of supports so some remain unbound, introducing supports into an array of wells, performing scanner-based well-level detection, and using a computer processor to determine the number of target nucleic acids per sample while assigning sample identity either from a distinguishable solid-support characteristic or from recorded well position. The variant coverage includes single-support-per-well methods using presence or absence, and multi-support-per-well methods using signal intensity in addition to presence or absence.
Stated Advantages
Enables sample identity determination by the detected characteristic of the solid support or by the detected position of the wells.
Enables determining the number of sample target nucleic acids per sample based on the number of wells having detected sample target nucleic acids.
Supports quantification using detected presence or absence and, in one claim variant, signal intensity across an array of wells using scanner detection and computer-processor analysis.
Documented Applications
No documented applications found
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