The medical field of COVID-19 diagnosis relates to methods for detecting SARS-CoV-2 nucleic acids in a sample as well as combinations of oligomers for determining the presence or absence of SARS-CoV-2 in a Sample.

The medical field of COVID-19 diagnosis relates to methods for detecting SARS-CoV-2 nucleic acids in a sample as well as combinations of oligomers for determining the presence or absence of SARS-CoV-2 in a Sample.

A method for generating region-specific amplification templates of a biological sample includes adding first oligonucleotide constructs and second oligonucleotide constructs to the biological sample. Each first or second oligonucleotide construct comprises a first or a second photoremovable cage molecule. The method further includes synthesising a complementary first strand from a template bound to target binding regions of each first oligonucleotide construct or each second oligonucleotide construct, scanning a first region of interest of the biological sample with a first focused light beam and a second region of interest of the biological sample with a second focused light beam to form uncaged first oligonucleotide constructs in the first region of interest and uncaged second oligonucleotide constructs in the second region of interest, synthesising a complementary second strand to form first amplification templates originating from the first region of interest and second amplification templates originating from the second region of interest.

A method for generating region-specific amplification templates of a biological sample includes adding first oligonucleotide constructs and second oligonucleotide constructs to the biological sample. Each first or second oligonucleotide construct comprises a first or a second photoremovable cage molecule. The method further includes synthesising a complementary first strand from a template bound to target binding regions of each first oligonucleotide construct or each second oligonucleotide construct, scanning a first region of interest of the biological sample with a first focused light beam and a second region of interest of the biological sample with a second focused light beam to form uncaged first oligonucleotide constructs in the first region of interest and uncaged second oligonucleotide constructs in the second region of interest, synthesising a complementary second strand to form first amplification templates originating from the first region of interest and second amplification templates originating from the second region of interest.

This invention generally relates to a novel RNA/mRNA production and amplification method using viral RNA replicase and/or RNA-dependent RNA polymerase (RdRp) enzymes as well as the associated mRNAs thereof. The present invention can be used for manufacturing and amplifying all varieties of RNA/mRNA sequences carrying at least an RdRp-binding site in the 5′- or 3′-end, or both. The RNA/mRNA so obtained is useful for not only producing mRNA vaccines and/or RNA-based medicines but also for generating the mRNA-associated proteins, peptides, and/or antibodies under an in-vitro as well as in-cell translation condition. Principally, the present invention is a novel RNA replicase-mediated RNA/mRNA amplification method, namely Replicase Cycling Reaction (RCR). The RNA replicases involved in RCR include but not limited to viral and/or bacteriophage RNA-dependent RNA polymerases (RdRp), particularly coronaviral and hepatitis C viral (HCV) RdRp enzymes.

This invention generally relates to a novel RNA/mRNA production and amplification method using viral RNA replicase and/or RNA-dependent RNA polymerase (RdRp) enzymes as well as the associated mRNAs thereof. The present invention can be used for manufacturing and amplifying all varieties of RNA/mRNA sequences carrying at least an RdRp-binding site in the 5′- or 3′-end, or both. The RNA/mRNA so obtained is useful for not only producing mRNA vaccines and/or RNA-based medicines but also for generating the mRNA-associated proteins, peptides, and/or antibodies under an in-vitro as well as in-cell translation condition. Principally, the present invention is a novel RNA replicase-mediated RNA/mRNA amplification method, namely Replicase Cycling Reaction (RCR). The RNA replicases involved in RCR include but not limited to viral and/or bacteriophage RNA-dependent RNA polymerases (RdRp), particularly coronaviral and hepatitis C viral (HCV) RdRp enzymes.

Provided herein is a method for detecting the presence of a COVID-19 virus RNA or other pathogenic respiratory viruses, such as an influenza virus, or other RNA of interest in a sample. Nucleic acids are obtained from the sample and are used as a template in a combined isothermal reverse transcription, RNAse H and isothermal amplification reaction to generate single stranded RNA amplicons containing sequences complementary to fluorescent labeled detector probes. The single-stranded RNA amplicons hybridize to the detector probe and to hybridization probes with sequences complementary to a sequence determinant in the COVID-19 or other virus RNAs. The microarray is imaged to detect fluorescent signals thereby identifying the virus.

Provided herein is a method for detecting the presence of a COVID-19 virus RNA or other pathogenic respiratory viruses, such as an influenza virus, or other RNA of interest in a sample. Nucleic acids are obtained from the sample and are used as a template in a combined isothermal reverse transcription, RNAse H and isothermal amplification reaction to generate single stranded RNA amplicons containing sequences complementary to fluorescent labeled detector probes. The single-stranded RNA amplicons hybridize to the detector probe and to hybridization probes with sequences complementary to a sequence determinant in the COVID-19 or other virus RNAs. The microarray is imaged to detect fluorescent signals thereby identifying the virus.

Recombinant transposon end nucleic acids are described that can incorporate barcodes, sequencing primers, or other functional biological sequences. This application also describes mixtures and uses of the recombinant transposon end nucleic acids.

Recombinant transposon end nucleic acids are described that can incorporate barcodes, sequencing primers, or other functional biological sequences. This application also describes mixtures and uses of the recombinant transposon end nucleic acids.