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.

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.

The invention relates to the technical field of nucleic acid detection and discloses a novel coronavirus nucleic acid rapid hybrid capture immunofluorescence detection kit, and a preparation method and a detection method thereof. The kit includes a COVID-19 reaction solution, wherein the COVID-19 reaction solution is prepared from a COVID-19 fluorescence marker and a COVID-19 probe solution; and the COVID-19 probe solution includes: an ORFlab section probe, an N section probe and an E section probe. Compared with general fluorescence PCR and sequencing detection, the kit has the advantages of stronger signal intensity, better specificity, shorter detection time, no need of professional technicians for operation, no need of refrigeration in transportation and storage, no need of a matched laboratory and a matched PCR instrument, and convenience and rapidness in use.

The invention relates to the technical field of nucleic acid detection and discloses a novel coronavirus nucleic acid rapid hybrid capture immunofluorescence detection kit, and a preparation method and a detection method thereof. The kit includes a COVID-19 reaction solution, wherein the COVID-19 reaction solution is prepared from a COVID-19 fluorescence marker and a COVID-19 probe solution; and the COVID-19 probe solution includes: an ORFlab section probe, an N section probe and an E section probe. Compared with general fluorescence PCR and sequencing detection, the kit has the advantages of stronger signal intensity, better specificity, shorter detection time, no need of professional technicians for operation, no need of refrigeration in transportation and storage, no need of a matched laboratory and a matched PCR instrument, and convenience and rapidness in use.

This specification discloses a novel methodology for labelling RNA via enzymatic incorporation of a minimally perturbing fluorescent tricyclic cytosine analogue. This analogue is shown to be 100% incorporated in example transcripts and is fully compatible with both in vitro and in cell transcription. Spectroscopic characterization shows that the incorporation rate of the cytosine analogue is on par with its natural counterpart. Using live cell imaging and flow cytometry, labelled mRNAs are efficiently and correctly translated upon transfection into living cells and cell-free systems. The spectral properties of the modified transcripts and their correct translation product allow for their straightforward and simultaneous visualization. This technology therefore offers a general route to understanding the biological behaviour of RNA of interest, including RNA based drugs. The fluorescent tricyclic cytosine analogue has formula (I):

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This specification discloses a novel methodology for labelling RNA via enzymatic incorporation of a minimally perturbing fluorescent tricyclic cytosine analogue. This analogue is shown to be 100% incorporated in example transcripts and is fully compatible with both in vitro and in cell transcription. Spectroscopic characterization shows that the incorporation rate of the cytosine analogue is on par with its natural counterpart. Using live cell imaging and flow cytometry, labelled mRNAs are efficiently and correctly translated upon transfection into living cells and cell-free systems. The spectral properties of the modified transcripts and their correct translation product allow for their straightforward and simultaneous visualization. This technology therefore offers a general route to understanding the biological behaviour of RNA of interest, including RNA based drugs. The fluorescent tricyclic cytosine analogue has formula (I):

##STR00001##

The present invention provides oligonucleotides and methods for their use in the detection and/or differentiation of target nucleic acids. The oligonucleotides and methods find particular application in amplifying, detecting, and/or discriminating multiple targets simultaneously.

The present invention provides oligonucleotides and methods for their use in the detection and/or differentiation of target nucleic acids. The oligonucleotides and methods find particular application in amplifying, detecting, and/or discriminating multiple targets simultaneously.

The present invention provides a method of quantification of a target nucleic acid, using at least any two of the genes SYT10, EPHA3, PLEKHF1 and KBTBD4 as control genes. In particular, the combination of the genes SYT10, EPHA3, PLEKHF1 and KBTBD4, known as the 4Plex, is provided as a control for nucleic acid quantification. The 4Plex has particular utility as a control for nucleic acid quantification by methylation-specific droplet digital PCR.

A method for matching a three-dimensional first image of at least one discrete entity with a three-dimensional second image of the at least one discrete entity is provided. The at least one discrete entity includes a biological sample and a plurality of constituent parts of a marker. The method includes: generating a first representation of the marker from the first image; generating a second representation of the marker from the second image; and based upon the representations matching, matching the first image with the second image; or based upon the representations not matching, rejecting the match. Generating the representations includes determining vectors from at least one reference item to at least some of the constituent parts of the marker, determining for the vectors at least one value of a property, and generating the representations of the marker based on a frequency of the at least one value of the property.