Unitary biochip providing sample-in to results-out processing and methods of manufacture

Inventors

Selden, Richard F. • Tan, Eugene

Assignees

Ande Corp

Abstract

A biochip for the integration of all steps in a complex process from the insertion of a sample to the generation of a result, performed without operator intervention includes microfluidic and macrofluidic features that are acted on by instrument subsystems in a series of scripted processing steps. Methods for fabricating these complex biochips of high feature density by injection molding are also provided.

Core Innovation

The invention provides an integrated, unitary, stationary biochip for conducting a multiplexed PCR analysis of at least two nucleic acids in an instrument that includes a process controller and a pneumatic subsystem. The biochip uses a pneumatic subassembly with a pneumatic plate aligned with a fluidics assembly, where drive lines contain only air and pressure is delivered to top ends of preloaded elution reagent storage chambers. The delivered pressure causes first and second foils to burst, and scripted controls from the process controller release contents from the preloaded reagent storage chambers.

The biochip includes a macrofluidic block with an inlet for receiving at least two nucleic acids and at least one preloaded elution reagent storage chamber that has a top end and a bottom end, with a first foil seal bonded to the bottom end and a second foil seal bonded to the top end. A fluidics subassembly is provided with at least one reconstitution chamber preloaded with lyophilized multiplexed PCR reagent and positioned in fluid communication with the elution reagent storage chamber, and at least one thermal cycling chamber. A primary flow path places the reconstitution chamber in fluidic communication with the thermal cycling chamber so that eluted nucleic acids are followed by reconstitution of the lyophilized multiplexed PCR reagent to generate PCR mix solution that travels along the flow path to the thermal cycling chamber.

In a unitary implementation, at least one combination reconstitution/thermal cycling chamber is preloaded with lyophilized multiplexed PCR reagent and positioned in fluid communication with the elution reagent storage chamber, allowing direct amplification after elution and reconstitution. A higher multiplexing architecture provides at least five preloaded elution reagent storage chambers, at least five reconstitution chambers, at least five thermal cycling chambers, and at least five primary flow paths to deliver PCR mix solutions to corresponding thermal cycling chambers.

Claims Coverage

The provided material includes three independent claims. Across these claims, the core inventive coverage centers on the pneumatic foil-burst release interface with scripted pressure control, the macrofluidic preloaded elution reagent chambers, and the microfluidic reconstitution and thermal cycling flow path arrangements for PCR mix generation and amplification.

The independent claims collectively require an instrument-driven pneumatic interface that bursts foil-sealed, preloaded elution reagent storage chambers under scripted controls, followed by microfluidic elution of nucleic acids and reconstitution of lyophilized multiplexed PCR reagent to generate PCR mix solution. They further require routing of the PCR mix to thermal cycling chambers, either via separate reconstitution and thermal cycling chambers, via direct amplification in a combination reconstitution/thermal cycling chamber, or via a higher multiplexing architecture providing at least five parallel sets of elution, reconstitution, and thermal cycling chambers and flow paths.

Stated Advantages

Documented Applications