The present invention provides, among other things, methods of quantitating mRNA capping efficiency, particularly mRNA synthesized in vitro. In some embodiments, methods according to the present invention comprise providing an mRNA sample containing capped and uncapped mRNA, providing a cap specific binding substance under conditions that permit the formation of a complex between the cap specific binding substance and the capped mRNA, and quantitatively determining the amount of the complex as compared to a control, thereby quantifying mRNA capping efficiency.

The embodiments disclosed herein utilized RNA targeting effectors to provide a robust CRISPR-based diagnostic with attomolar sensitivity. Embodiments disclosed herein can detect both DNA and RNA with comparable levels of sensitivity and can differentiate targets from non-targets based on single base pair differences. Moreover, the embodiments disclosed herein can be prepared in freeze-dried format for convenient distribution and point-of-care (POC) applications. Such embodiments are useful in multiple scenarios in human health including, for example, viral detection, bacterial strain typing, sensitive genotyping, and detection of disease-associated cell free DNA.

The embodiments disclosed herein utilized RNA targeting effectors to provide a robust CRISPR-based diagnostic with attomolar sensitivity. Embodiments disclosed herein can detect both DNA and RNA with comparable levels of sensitivity and can differentiate targets from non-targets based on single base pair differences. Moreover, the embodiments disclosed herein can be prepared in freeze-dried format for convenient distribution and point-of-care (POC) applications. Such embodiments are useful in multiple scenarios in human health including, for example, viral detection, bacterial strain typing, sensitive genotyping, and detection of disease-associated cell free DNA.

The present invention relates to the detection of target nucleic acids using enzyme-assisted nanotechnology. More specifically, the present invention provides a molecular nanotechnology in the form of a transition-state DNA-enzyme molecular switch and methods of use that enables direct and sensitive detection of viral RNA targets in native clinical samples. In one embodiment, the detection comprising steps of providing a composition comprising at least one DNA polymerase enzyme, at least one enhancer, and at least one DNA polymerase inhibitor, wherein the DNA polymerase inhibitor is recognized and bound by the DNA polymerase enzyme via its conserved region, and is complementary to a portion of the enhance via its variable region. In another embodiment, the detection method comprising steps of providing a signalling nanostructure and detecting signal development, wherein a change in the intensity of signal. In an alternative embodiment, the target nucleic acid is a SARS-CoV-2 polynucleotide.

The present invention relates to the detection of target nucleic acids using enzyme-assisted nanotechnology. More specifically, the present invention provides a molecular nanotechnology in the form of a transition-state DNA-enzyme molecular switch and methods of use that enables direct and sensitive detection of viral RNA targets in native clinical samples. In one embodiment, the detection comprising steps of providing a composition comprising at least one DNA polymerase enzyme, at least one enhancer, and at least one DNA polymerase inhibitor, wherein the DNA polymerase inhibitor is recognized and bound by the DNA polymerase enzyme via its conserved region, and is complementary to a portion of the enhance via its variable region. In another embodiment, the detection method comprising steps of providing a signalling nanostructure and detecting signal development, wherein a change in the intensity of signal. In an alternative embodiment, the target nucleic acid is a SARS-CoV-2 polynucleotide.

This disclosure provides a method of determining a sequence of nucleotides for a nucleic acid template. The method can include the steps of contacting the nucleic acid template with a conformationally labeled polymerase and at least four different nucleotide species under conditions wherein the conformationally labeled polymerase catalyzes sequential addition of the nucleotide species to form a nucleic acid complement of the nucleic acid template, wherein the sequential addition of each different nucleotide species produces a conformational signal change from the conformationally labeled polymerase and wherein the rate or time duration for the conformational signal change is distinguishable for each different nucleotide species; detecting a series of changes in the signal from the conformationally labeled polymerase under the conditions; and determining the rates or time durations for the changes in the signal, thereby determining the sequence of nucleotides for the nucleic acid template.

This disclosure provides a method of determining a sequence of nucleotides for a nucleic acid template. The method can include the steps of contacting the nucleic acid template with a conformationally labeled polymerase and at least four different nucleotide species under conditions wherein the conformationally labeled polymerase catalyzes sequential addition of the nucleotide species to form a nucleic acid complement of the nucleic acid template, wherein the sequential addition of each different nucleotide species produces a conformational signal change from the conformationally labeled polymerase and wherein the rate or time duration for the conformational signal change is distinguishable for each different nucleotide species; detecting a series of changes in the signal from the conformationally labeled polymerase under the conditions; and determining the rates or time durations for the changes in the signal, thereby determining the sequence of nucleotides for the nucleic acid template.

A method for testing a sample for the presence of one or more targets comprises multiplexed catalyzed reporter deposition (CARD) is provided.

A method for testing a sample for the presence of one or more targets comprises multiplexed catalyzed reporter deposition (CARD) is provided.

Kits, compositions, and methods for labeling target materials or target analytes are considered. The composition can include one or more affinity molecules indirectly conjugated to one or more detection moieties. The kit can be used to detect a single biomarker or multiple biomarkers. The kit can be used to perform a single round of labeling or multiple rounds of labeling.