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Publication Number
US-9174210-B2
Publication Date
2015-11-03
Expiration Date
Abstract
A self-contained apparatus for isolating nucleic acid, cell lysates and cell suspensions from unprocessed samples apparatus, to be used with an instrument, includes at least one input, and: (i) a macrofluidic component, including a chamber for receiving an unprocessed sample from a collection device and at least one filled liquid purification reagent storage reservoir; and (ii) a microfluidic component in communication with the macrofluidic component through at least one microfluidic element, the microfluidic component further comprising at least one nucleic acid purification matrix; and (iii) at least one interface port to a drive mechanism on the instrument for driving said liquid purification reagent, through the microfluidic element and the nucleic acid purification matrix, wherein the only inputs to the apparatus are through the chamber and the interface port to the drive mechanism.
Core Innovation
The invention is a self-contained apparatus for isolating nucleic acid from an unprocessed sample. It combines macrofluidic and microfluidic components, pre-filled reagent storage chambers, a nucleic acid purification matrix, and a drive line that interfaces with an instrument. The instrument supplies controlled flow, pressure, or volumetric displacement of gas or liquid to move reagent through the microfluidic element and purification matrix.
A related apparatus generates a cell lysate from an unprocessed sample using macrofluidic and microfluidic components, pre-filled reagent chambers, a purification matrix, and an instrument-driven interface. The apparatus processes samples without intermediate manual preparation and is suitable for field and point-of-care use. Fluidic configurations include macrofluidic chambers, microfluidic channels, and an integrated matrix region.
Claims Coverage
The claim coverage includes four inventive features across two independent claims directed to nucleic acid isolation and cell lysate generation.
Self-contained nucleic acid isolation apparatus
The first independent claim covers an apparatus for isolating nucleic acid from an unprocessed sample, comprising macrofluidic and microfluidic components, pre-filled reagent storage chambers, a nucleic acid purification matrix, drive lines, and an instrument interface that supplies controlled flow, pressure, or volumetric displacement.
Instrument-driven reagent movement
The apparatus has a drive mechanism interface configured to drive liquid purification reagent through a microfluidic element and nucleic acid purification matrix and back into another macrofluidic chamber. The drive line supplies controlled flow, controlled pressure, or controlled volumetric displacement of gas or liquid.
Self-contained cell lysate generation apparatus
The second independent claim covers a self-contained apparatus for generating a cell lysate from an unprocessed sample, using macrofluidic and microfluidic components, pre-filled reagent storage chambers, a nucleic acid purification matrix, drive lines, and an instrument interface that supplies controlled flow, pressure, or volumetric displacement.
Reagent chambers and purification matrix
Both independent claims specify reagent storage chambers pre-filled with necessary reagents and a microfluidic component containing at least one nucleic acid purification matrix. The second claim specifies that the apparatus generates a cell lysate; its dependent claims identify a lysis reagent and gas bubbling to stir the mixture and lyse cells.
The two independent claims cover instrument-connected, self-contained apparatus configurations for nucleic acid isolation and cell lysate generation, both defined by macrofluidic and microfluidic components, pre-filled reagent storage chambers, a purification matrix, and controlled gas or liquid movement through an instrument interface.
Stated Advantages
The apparatus processes unprocessed samples without intermediate manual preparation.
The apparatus is described as suitable for field and point-of-care use.
The apparatus is described as suitable for integration with downstream analytical instruments.
Documented Applications
The apparatus is described for isolating nucleic acids from unprocessed samples and for generating cell lysates or cell suspensions.
The apparatus is described for processing unprocessed clinical, forensic, and environmental samples, including whole blood and forensic samples such as buccal swabs, bloodstains, saliva, and touch samples.
The apparatus is described for use with downstream analytical instruments for techniques including thermal cycling, capillary or microfluidic electrophoresis, nucleic acid fragment sizing, STR and related analyses, PCR, sequencing, hybridization, microarrays, immunoassays, mass spectroscopy, and RFLP analyses, with STR electropherogram profiles depicted across multiple loci.
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