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Publication Number
US-10618047-B2
Publication Date
2020-04-14
Expiration Date
Abstract
This disclosure relates to an apparatus for simultaneously filling a plurality of sample chambers. In one aspect, the apparatus comprises a common fluid source and a plurality of independent, continuous fluidic pathways. Each independent, continuous fluidic pathway comprises a sample chamber and a pneumatic compartment. The sample chamber is connected to the common fluid source, and the pneumatic compartment is connected to the sample chamber. The sample chamber comprises, in part, an assay chamber. The assay chamber comprises a monolithic substrate and a plug. In some embodiments, the assay chamber contains a magnetic mixing element. In some embodiments, the assay chamber is a double tapered chamber. In some embodiments, a ratio of a volume of the sample chamber to a volume of the pneumatic compartment is substantially equivalent for each fluidic pathway of the plurality of fluidic pathways.
Core Innovation
The microfluidic assay device and method provide an apparatus with a common fluid source containing a fluid sample comprising a nucleic acid, and a plurality of independent, continuous fluidic pathways. Each independent, continuous fluidic pathway includes a sample chamber connected to the common fluid source and a pneumatic compartment connected to the sample chamber, and each pathway is a closed system excluding the connection between the sample chamber and the common fluid source.
Concurrent filling is achieved by selecting a ratio of the fluid volume to the pneumatic volume that is substantially equivalent for each fluidic pathway of the plurality of fluidic pathways. Applying a supply pressure to the common fluid source causes the fluid sample to flow into the sample chamber of each fluidic pathway to substantially fill the sample chamber, and heating each sample chamber amplifies one or more nucleic acids in the fluid sample while maintaining the closed pathway environment.
The sample chamber comprises an assay chamber having a first bounding surface formed in a monolithic substrate and a second bounding surface formed by a transparent plug. The plug includes a body and a cap, where the body protrudes into the monolithic substrate at a depth, and the cap forms the second bounding surface of the assay chamber, supporting visual detection of amplification through the transparent plug.
Claims Coverage
The independent claim in the partial content is clm-00001, which defines a nucleic-acid amplification detection method using a microfluidic apparatus with a common fluid source, multiple independent, continuous closed fluidic pathways each with a sample chamber and a pneumatic compartment, substantially equivalent fluid volume-to-pneumatic volume ratios for substantially simultaneous filling, and amplification by heating with visual detection through a transparent plug. The dependent refinements specify ramping supply pressure, sealing after filling before heating, a plug cap internal cavity containing dried reagents, and amplification by LAMP.
Common fluid source to multiple independent closed pathways
An apparatus comprising a common fluid source containing a fluid sample comprising a nucleic acid, and a plurality of independent, continuous fluidic pathways, each fluidic pathway comprising a sample chamber connected to the common fluid source and a pneumatic compartment connected to the sample chamber, wherein each fluidic pathway is a closed system excluding the connection between the sample chamber and the common fluid source.
Substantially equivalent fluid volume-to-pneumatic volume ratio
A ratio of the fluid volume to the pneumatic volume is substantially equivalent for each fluidic pathway of the plurality of fluidic pathways, so that applying a supply pressure fills each sample chamber substantially simultaneously.
Monolithic substrate assay chamber with transparent plug bounding surface
Each sample chamber is an assay chamber having a first bounding surface formed in a monolithic substrate and a second bounding surface formed by a transparent plug, the plug comprising a body and a cap, where the body protrudes into the monolithic substrate at a depth and the cap forms the second bounding surface of the assay chamber.
Visual detection through transparent plug after heating amplification
Applying a supply pressure to the common fluid source so the fluid sample flows into the sample chamber of each fluidic pathway to substantially fill the sample chamber, heating each sample chamber to amplify one or more nucleic acids in the fluid sample, and visually detecting amplification of nucleic acid through the transparent plug.
Ramping supply pressure for filling
Ramping the supply pressure.
Sealing each closed fluidic pathway after substantially filling and before heating
Sealing the fluidic pathway after substantially filling the sample chamber of each fluidic pathway and before heating the sample chamber.
Transparent plug cap with internal cavity and dried reagents
Providing a plug cap having an internal cavity that contains one or more dried reagents.
Amplification using LAMP
Amplifying one or more nucleic acids in the fluid sample is performed using LAMP.
Across the independent claim and the identified dependent refinements, the inventive concept is a closed microfluidic system with a common fluid source feeding multiple independent continuous fluidic pathways that have substantially equivalent fluid volume-to-pneumatic volume ratios for substantially simultaneous filling, monolithic-substrate assay chambers sealed with a transparent plug for visual readout, and amplification by heating followed by visually detecting amplification through the transparent plug, with refinements specifying ramped supply pressure, sealing after filling and prior to heating, plug cap internal cavities with dried reagents, and LAMP amplification.
Stated Advantages
Reduced bubble formation supported by a sealed pressure state enabled by sealing the closed fluidic pathway after substantially filling.
Reliable assays supported by a sealed pressure state in closed fluidic pathways.
Visual detection of nucleic-acid amplification through the transparent plug.
Documented Applications
Detecting amplified nucleic acid by visually detecting amplification through a transparent plug after heating an assay chamber in a microfluidic device using multiple independent, continuous closed fluidic pathways fed from a common fluid source.
Nucleic-acid amplification and detection compatible with LAMP, including visual detection through the transparent plug.
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