Abstract: Embodiments disclosed herein include devices, methods, and systems for direct, selective, and sensitive detection of single-stranded and double-stranded target DNA sequences from various sources using a Cas12a protein. When activated by binding a target DNA sequence, the Cas12a cleaves a tether releasing a reporter molecule that may then be detected. In some embodiments, the systems, methods, and devices may include a filter or membrane that may help to separate the tethered and untethered reporter molecules. These devices, systems, and techniques allow a user to rapidly process samples that may contain the target DNA, without needing to amplify the target sequences. These devices and methods may be used to assay a wide variety of samples and target DNA sources, for the presence or absence of target DNA sequences. Compositions and kits, useful in practicing these methods, for example detecting a target DNA in a biological sample, are also described.

Abstract: Embodiments disclosed herein include devices, methods, and systems for direct, selective, and sensitive detection of single-stranded and double-stranded target nucleic acid sequences from various sources in a solution-based system. When activated by binding a target nucleic acid sequence, the Cas protein cleaves a tether separating a reporter molecule from a capture moiety. The capture moiety can then be used to remove, localize, or sequester uncleaved molecule containing intact tethers. In some embodiments, the systems, methods, and devices may include a filter, a membrane, or other molecules that may help to separate the tethered and untethered reporter molecules and/or capture the reporter molecules. These devices, systems, and techniques allow a user to rapidly process samples that may contain the target nucleic acid, in some cases, without needing to amplify the target sequences, and without the need for sophisticated or expensive laboratory equipment.

Abstract: Generally, this disclosure relates to expression constructs that encode a reporter enzyme-affinity binding tag fusion protein that is produced after the construct is inserted into bacteriophage and the bacteriophage infects bacteria. In some embodiments, the fusion protein is captured and produces a detectable signal. Signal intensity may correlate with the number of bacterial cells in a fluid sample. Methods of detecting bacteria using the expression constructs, and microfluidic devices for detecting bacteria using the expression constructs are also disclosed.

Abstract: Controlling a valveless microfluidic device using rotational and pistoning motion is disclosed. A system embodiment includes, but is not limited to, a stage to support a valveless microfluidic device, the stage having a bottom surface coupled with an extension, having a threaded portion, and defining an interior volume; a first motor including a powered rotating portion operable to engage with the threaded portion to induce a z-axis motion of the stage; a second motor including a powered rotating portion coupled with a slidable coupler, the slidable coupler engaging the interior volume to induce a second motion of the stage about the z-axis; a base defining an aperture to receive the stage; and a lid having a subset of the plurality of apertures corresponding to a subset of a plurality of apertures defined by the valveless microfluidic device upon positioning of the stage by the first motor and the second motor.

Abstract: Embodiments disclosed herein include devices, methods, and systems for direct, selective, and sensitive detection of single-stranded and double-stranded target DNA sequences from various sources using a programmed Cas12a protein. When activated by binding a target DNA sequence, the Cas12a cleaves a tether releasing a reporter molecule that may then be detected. In some embodiments, the systems, methods, and devices may include a filter or membrane that may help to separate the tethered and untethered reporter molecules. These devices, systems, and techniques allow a user to rapidly process samples that may contain the target DNA, without needing to amplify the target sequences. These devices and methods may be used to assay a wide variety of samples and target DNA sources, for the presence or absence of a specific target DNA sequences. Compositions and kits, useful in practicing these methods, for example detecting a target DNA in a biological sample, are also described.

Abstract: Embodiments disclosed herein include devices, methods, and systems for direct, selective, and sensitive detection of single-stranded target RNA sequences from various sources using a programmed Cas13a protein. When activated by binding a target RNA sequence, the Cas13a cleaves a tether releasing a reporter molecule that may then be detected. In some embodiments, the systems, methods, and devices may include a filter or membrane that may help to separate the tethered and untethered reporter molecules. These devices, systems, and techniques allow a user to rapidly process samples that may contain the target RNA, without needing to amplify the target sequences. These devices and methods may be used to assay a wide variety of samples and target RNA sources, for the presence or absence of a specific target RNA sequence. Compositions and kits, useful in practicing these methods, for example detecting a target RNA in a biological sample, are also described.

Abstract: Generally, this disclosure relates to expression constructs that encode a reporter enzyme-affinity binding tag fusion protein that is produced after the construct is inserted into bacteriophage and the bacteriophage infects bacteria. In some embodiments, the fusion protein is captured and produces a detectable signal. Signal intensity may correlate with the number of bacterial cells in a fluid sample. Methods of detecting bacteria using the expression constructs, and microfluidic devices for detecting bacteria using the expression constructs are also disclosed.

Abstract: A microfluidic device for concentrating and detecting bacteria in liquids, and related methods are described. The device includes a first filter chamber for capturing bacteria and performing incubations of the bacteria with one or more reagents, and a second filter chamber for capturing and concentrating a detectable material, with little or no binding of detectable material by the first filter. In an aspect, bacteria are incubated with growth media and engineered phage that cause the bacteria to produce an enzyme. In an aspect, the enzyme is capture in the second filter chamber and exposed to a substrate to produce a detectable signal.