Omega amplification

Inventors

Wang, Youxiang • Chen, Xin • Wang, Rong • Yang, Zhijie • Zhao, Yu

Assignees

Atila BioSystems Inc

Abstract

The present disclosure provides compositions, methods and kits for Omega amplification technologies. In addition, the present disclosure provides compositions, methods and kits for universal FQ probe and for G-quadruplex detection methods for use in isothermal amplification technologies.

Core Innovation

An omega amplification primer set is provided for isothermal amplification of a portion of a target nucleic acid sequence. The primer set includes a first foldback primer and a second foldback primer under suitable omega amplification conditions. At least one of the foldback primers comprises an extruding sequence at its 5′ terminus, where the extruding sequence is at least 2 nucleotides in length and does not anneal to a template nucleic acid or hybridize to the first strand or a complementary strand of the target nucleic acid sequence.

The extruding sequence-based design is used within foldback primer amplification, with a 5′ extruding sequence that does not hybridize to the downstream of the same strand. This arrangement prevents a particular 3′-OH extension mode while maintaining near-LAMP speed within omega amplification using foldback primers. Experimental comparisons to LAMP are described, including gel and restriction-digest behavior showing non-repetitive extruding sequence duplication.

Universal nucleic acid detection systems are also described for real-time monitoring of isothermal strand-displacing amplification. These include universal fluorescent quencher probes that comprise fluorophore and quencher complementary oligonucleotides activated by a specific detection probe, including invader/invader-kicker architectures and optional activation modes. Additional detection is described using structural-feature motifs, including G-quadruplex or aptamer motifs formed during amplification, to generate exponential signal generation in real-time monitoring formats.

Claims Coverage

The document identifies one independent claim for the omega amplification primer set, with additional dependent claims refining primer features and optional accelerator and detection-related sequence types. The inventive features are centered on the foldback primer extruding sequences at the 5′ terminus, including constraints on length and hybridization and annealing behavior.

Claim coverage is grounded in extruding-sequence foldback primer design for omega amplification, requiring 5′ extruding sequences of at least 2 nucleotides that do not anneal or hybridize to the target’s first or complementary strands, thereby enabling isothermal omega amplification of a portion of the target nucleic acid sequence under suitable omega amplification conditions.

Stated Advantages

Documented Applications