Devices and methods for nucleic acid identification in samples

A method for monitoring fluorescence associated with nucleic acid material in a sample, the method comprising:

• providing a microfluidics system;
• the microfluidics system including a movable cartridge assembly;
• the movable cartridge assembly comprising:
• a surface configured to receive the sample;
• at least one target-specific set of reagent components deposited on the surface at a target location;
• at least a second target-specific set of reagent components deposited on the surface at a second target location;
• at least one set of RCA-LAMP reaction components deposited on the surface at an RCA-LAMP location;
• the at least one target-specific set of reagent components including:
• at least one target-specific padlock probe reagent component;
• at least one target probe-associated ligase enzyme component;
• and
• at least one target probe-associated set of ligase buffer components;
• the second target-specific set of reagent components including:
• a second target-specific padlock probe reagent component;
• the at least one target probe-associated ligase enzyme component;
• and
• the at least one target probe-associated set of ligase buffer components;
• and
• the at least one set of RCA-LAMP reaction components including:
• at least one polymerase buffer component;
• at least one polymerase enzyme with strand displacement activity;
• a betaine additive;
• a poloxamine surfactant additive;
• a sequence-specific probe;
• dNTPs;
• and
• a primer mix;
• the primer mix including:
• a forward inside primer specific to a backbone of the at least one target-specific padlock probe reagent component;
• and
• a backward inside primer specific to the backbone of the at least one target-specific padlock probe reagent component;
• wherein the second target-specific padlock probe reagent component includes the backbone of the at least one target-specific padlock probe reagent component;
• transporting an aliquot of the sample received on the surface to the target location;
• transporting a second aliquot of the sample received on the surface to the second target location;
• applying heat to the target location and the second target location;
• transporting an aliquot of RCA-LAMP reaction components on the surface to the target location;
• transporting a second aliquot of RCA-LAMP reaction components on the surface to the second target location;
• applying heat to the target location and the second target location;
• providing excitation radiation to the target location and the second target location;
• and
• monitoring fluorescence emanating from the target location and the second target location in response to the excitation radiation using a camera system.