The relative abundance of bacterial species in a patient’s microbiome is quantified using DNA nanostructures that fluoresce multiple colors. Immobilizing binders have binding sites with nucleotide sequences complementary to those at a primary site on rRNA subunits of each selected bacterial species. Fluorophore binders have binding sites with nucleotide sequences complementary to those at a secondary site on the rRNA subunits. The fluorophore binders for each bacterial species are attached to nanostructures that fluoresce a particular color for each bacteria. The immobilizing binders are attached to the surface of a microscopy chamber. RNA subunits are extracted from a microbiome sample of the patient and are attached to the corresponding immobilizing binders and fluorophore binders such that the RNA subunits of each bacterial species fluoresce a color unique to the species. DNA nanostructures emitting the same color are counted to determine the relative concentration of the bacterial species in the sample.

Described herein are DNA-nanostructures that can be used in an assay to detect and/or quantify an analyte of interest. Aspects of the DNA-nanostructure can include a single DNA molecule composed of hairpin structural motifs, an anchor recognition moiety, and a signal moiety, where the anchor recognition moiety and the signal moiety are in effective proximity to each other such that the tethered diffusion of the signal molecule can be altered based upon binding status of the anchor recognition moiety. Also described herein are methods of making and using the DNA-nanostructures.

Described herein are DNA-nanostructures that can be used in an assay to detect and/or quantify an analyte of interest. Aspects of the DNA-nanostructure can include a single DNA molecule composed of hairpin structural motifs, an anchor recognition moiety, and a signal moiety, where the anchor recognition moiety and the signal moiety are in effective proximity to each other such that the tethered diffusion of the signal molecule can be altered based upon binding status of the anchor recognition moiety. Also described herein are methods of making and using the DNA-nanostructures.

The invention provides methods, compositions, kits and devices for the detection of target molecules. In some embodiments, the invention allows for multiplexed target molecule detection.

The invention provides methods, compositions, kits and devices for the detection of target molecules. In some embodiments, the invention allows for multiplexed target molecule detection.

The present invention pertains to an enzymatic measurement method using an antibody-enzyme complex or a nucleic acid probe measurement method using an enzyme-labeled nucleic acid probe, in both of which the quantification of a product of a reaction by an enzyme in the antibody-enzyme complex or the enzyme-labeled nucleic acid probe is performed by generating thio-NAD(P)H by an enzymatic cycling reaction using NAD(P)H, thio-NAD(P), and a dehydrogenase (DH), and measuring the amount of the generated thio-NAD(P)H or measuring a change in color caused by the generated thio-NAD(P)H. An enzymatic reaction system in which NAD(P) generated from NAD(P)H by the enzymatic cycling reaction is selectively reduced, is caused to coexist with the enzymatic cycling reaction. The present invention also pertains to a kit for enzyme immunoassay, and a kit for nucleic acid probe measurement. In the enzymatic cycling reaction, the detection sensitivity is increased by increasing the amount of thio-NAD(P)H generated per unit time with respect to a predetermined amount of a substrate (reduced), and combining the same with an enzyme immunoassay, etc., enables quantification, etc., of a protein or nucleic acid with high sensitivity.

The present invention pertains to an enzymatic measurement method using an antibody-enzyme complex or a nucleic acid probe measurement method using an enzyme-labeled nucleic acid probe, in both of which the quantification of a product of a reaction by an enzyme in the antibody-enzyme complex or the enzyme-labeled nucleic acid probe is performed by generating thio-NAD(P)H by an enzymatic cycling reaction using NAD(P)H, thio-NAD(P), and a dehydrogenase (DH), and measuring the amount of the generated thio-NAD(P)H or measuring a change in color caused by the generated thio-NAD(P)H. An enzymatic reaction system in which NAD(P) generated from NAD(P)H by the enzymatic cycling reaction is selectively reduced, is caused to coexist with the enzymatic cycling reaction. The present invention also pertains to a kit for enzyme immunoassay, and a kit for nucleic acid probe measurement. In the enzymatic cycling reaction, the detection sensitivity is increased by increasing the amount of thio-NAD(P)H generated per unit time with respect to a predetermined amount of a substrate (reduced), and combining the same with an enzyme immunoassay, etc., enables quantification, etc., of a protein or nucleic acid with high sensitivity.

Provided herein is a reverse transcriptase mixture comprising a reverse transcriptase and a colored dye at a concentration in the range of 0.003%-1% (v/w). The colored dye may be visually observed during transfer of the mix from one vessel to another and addition of the mix to another mix can be confirmed by eye by observing the colored dye.

Provided herein is a reverse transcriptase mixture comprising a reverse transcriptase and a colored dye at a concentration in the range of 0.003%-1% (v/w). The colored dye may be visually observed during transfer of the mix from one vessel to another and addition of the mix to another mix can be confirmed by eye by observing the colored dye.

The invention discloses a composition and kit for early detection of high-grade cervical lesions and cervical cancer, wherein the composition for early detection of high-grade cervical lesions and cervical cancer includes methylation primers, a probe corresponding to methylated sites and methylation blocking primers for FAM19A4 gene; methylation primers, a probe corresponding to methylated sites and methylation blocking primers for JAM3 gene; methylation primers, a probe corresponding to methylated sites and methylation blocking primers for PAX1 gene; and 1 pair of primers and a probe corresponding to methylated sites for internal reference gene GAPDH. The methylated sites in FAM19A4, JAM3 and PAX1 genes are accurately detected using multiple multi-channel fluorescence and blocking techniques through accurate recognition between specific primers and probes and methylated sequences, full release of methylated templates under the action of multiple blocking primers and optimized special methylation DNA polymerase.