Blocked Nucleic Acid Cascade Assay for Rapid Target Detection

The patent extracts and eliminates the amplification step from the detection process. By using a cascade assay with two ribonucleoprotein complexes and blocked nucleic acid molecules, the system achieves detection sensitivity without requiring target nucleic acid amplification, thus reducing detection time and avoiding artifacts caused by amplification.

The patent introduces blocked nucleic acid molecules as intermediaries that mediate the detection process. These blocked molecules are cleaved by the first ribonucleoprotein complex when bound to the target, releasing unblocked molecules that activate the second complex, enabling signal amplification without target nucleic acid amplification.

The patent removes the amplification step that causes artifacts. By using a direct detection cascade assay with blocked nucleic acid molecules, the system achieves high sensitivity without the artifacts and inaccuracies introduced by conventional nucleic acid amplification methods.

The patent converts the blocking function of blocked nucleic acid molecules from a hindrance into a beneficial mechanism. The blocked molecules are designed to be cleaved only when the target nucleic acid is present, transforming the blocking state into a controlled signal activation mechanism that improves detection accuracy.

The patent segments the detection process into two distinct ribonucleoprotein complexes with specialized functions. The first complex handles target binding and initial cleavage, while the second complex provides signal amplification, enabling rapid detection through functional segmentation rather than simple sequential steps.

The patent employs preliminary action by pre-assembling the two ribonucleoprotein complexes and blocked nucleic acid molecules before target introduction. This pre-preparation allows the cascade reaction to proceed rapidly once the target is added, achieving fast detection without requiring complex real-time assembly steps.