85 results about "Locked nucleic acid" patented technology

The invention relates to the technical field of pathogenic bacterium detection, in particular to a method and a kit for detecting helicobacter pylori gene mutation in excrement. According to the technical scheme, the kit for detecting helicobacter pylori gene mutation in excrement is developed by combining multiple advanced technical means such as ARMS-PCR design, blocking primers, locked nucleic acid modification (peptide nucleic acid modification), MGB probes, nested PCR design and degenerate base design. The kit can detect nine gene mutation sites related to the drug resistance of helicobacter pylori at one time, and has a wide coverage range. The kit in the scheme can be used for detecting the drug resistance of quinolone antibiotics and / or clarithromycin antibiotics of helicobacter pylori, and the technical problem that a high-specificity and high-sensitivity excrement helicobacter pylori gene mutation detection method is lacked in the prior art is solved. The method is simple and convenient to operate and can be completed within 1-2 hours, the detection cost is reduced, and the method has ideal application and popularization prospects.

The invention relates to the technical field of biology, in particular to a cis-trans typing method for drug-resistant gene mutation EGFR T790M / C797S. The invention provides an EGFR (Epidermal Growth Factor Receptor) T790M / C797S cis-trans typing method based on a locked nucleic acid LNA (Low Nucleic Acid) probe, ultrahigh-specificity DNA (Deoxyribose Nucleic Acid) polymerase and a primer probe combination. The typing method can be used for accurately distinguishing EGFR T790M / C797S double-mutation cis-configuration and trans-configuration; meanwhile, the kit has the advantages of high sensitivity, high specificity, simple steps and low cost, is suitable for detecting low-abundance samples such as circulating tumor DNA (ctDNA) and can meet the clinical rapid detection requirement.

According to the present invention, the compound represented by the formula I is synthesized through the brand new nucleoside intermediate, the compound represented by the formula I is the important precursor nucleoside # imgabs0 #, and the definition of R1 is as defined in the specification. The preparation method provided by the invention overcomes the defects of more reaction byproducts and low yield in the prior art, and has a better application prospect.

The invention belongs to the field of molecular biological detection, and particularly relates to combined nucleic acid detection of three typical mosquito-borne arthroviruses, namely, dengue virus, Zika virus and Chikungunya virus, and four serotypes of the dengue virus are typed by using a melting curve matched molecular beacon design method. The method specifically comprises the following steps: distinguishing and detecting three typical mosquito-borne arbo viruses by using an amplification curve, and marking pathogens in each tube on different probes by using different fluorophores so as to distinguish amplification signals. A melting curve is used for distinguishing four different types of viruses of the dengue virus, and Tm value characteristic peak signals are distinguished by combining a molecular beacon and a locked nucleic acid probe aiming at each type of specific target. According to the invention, clinical differential diagnosis of the three viruses is facilitated, and an auxiliary means is provided for guiding medication for different clinical manifestations in dengue virus typing detection.

The invention provides a nucleic acid probe, a primer group, a kit, a detection method and application, and relates to the technical field of biology. The nucleotide sequence of the nucleic acid probe is as shown in SEQ ID NO. 1; wherein the 3'end of the nucleic acid probe is modified with MGB, and locked nucleic acid is modified at a base position corresponding to the difference between the swine fever wild strain and the swine fever vaccine strain in the nucleotide sequence. According to the nucleic acid probe, a specific sequence is utilized to precisely target a swine fever wild strain, and locked nucleic acid modification is introduced at a differential site, so that the recognition capability on single-base mismatch is remarkably enhanced, non-specific binding to a vaccine strain is thoroughly blocked, and false positive interference is eliminated; and 3 '-terminal MGB modification is matched to improve the binding stability and sensitivity, so that the efficient and accurate identification of the swine fever wild strain under the immune background is ensured.

The preparation method of the locked nucleic acid intermediate D-ribofuranose derivative, provided by the invention, is simple in steps, stable in process, high in selectivity and easy for industrial production, and has a relatively good application prospect. According to the preparation method, the yield and the purity can be greatly improved, the economic cost is reduced, steps are shortened, multiple operation modes are simplified, the amount of organic waste liquid is remarkably reduced, materials and process conditions with safety risks are eliminated, operation with serious equipment loss is avoided, and the process conditions and the result are stable.

The present invention relates to hybrid full LNA oligonucleotides. The present report relates to hybridizing single stranded (ss-) oligonucleotides consisting entirely of locked nucleic acid (LNA) monomers. This document shows hybridization experiments of pairs of fully complementary ss-oligonucleotides that fail to form double strands within a given time interval. The present report provides a method of identifying such incompatible oligonucleotide pairs. In another aspect, the present report provides pairs of complementary ss-oligonucleotides capable of rapidly forming double strands. The present report also provides methods of identifying and selecting compatible oligonucleotide pairs. In another aspect, the present report provides the use of a compatible oligonucleotide pair as a binding partner in binding assays, such as receptor-based assays.

The invention discloses a primer group for identifying phthorimaea operculella based on ddPCR and a method for identifying phthorimaea operculella by detecting environmental DNA, and belongs to the technical field of pest detection. The primer group for identifying the phthorimaea operculella based on the ddPCR comprises a forward primer, a reverse primer and a specific fluorescent probe, the nucleic acid sequences of the forward primer, the reverse primer and the specific fluorescent probe are as shown in SEQ ID NO.1-SEQ ID NO.3; the C basic group at the 3'end of the forward primer, the A basic group at the 3 'end of the reverse primer, and the A basic groups and the T basic groups at the 12th site and the 13th site of the specific fluorescent probe are basic groups modified by locked nucleic acid. The primer group disclosed by the invention has very high specificity and sensitivity to the phthorimaea operculella, can be detected by taking eDNA as a template, can find the phthorimaea operculella earlier, and has an important application prospect in early detection and prevention of the phthorimaea operculella.

The invention provides a primer probe, a kit and a method for identifying A1 / A2-beta casein in milk and dairy products. The primer probe comprises an allele specific primer A1A2-F, an allele specific primer A1A2-R, a probe A2-P and a probe A1-P. The invention further discloses a kit for detecting the allele specific primer A1A2-F, the allele specific primer A1A2-R and the probe A1-P. According to the present invention, based on the 67th single nucleotide polymorphism (SNP) of the ruminant A1 / A2-beta casein gene (CSN2), the gene typing method with characteristics of high specificity and high sensitivity is constructed; according to the method, an ARMS-PCR technology, a locked nucleic acid (LNA) modification strategy and an MGB probe signal detection system are integrated, so that the A1 / A2 allele is accurately distinguished.

Provided are compositions and methods of treating Coronavirus disease 2019 (COVID-19) in a subject, the method including administering to the subject a therapeutically effective amount of a composition comprising an exogenous nucleic acid and delivering the exogenous nucleic acid into a cell, wherein the exogenous nucleic acid comprises an antisense oligonucleotide, a small interfering RNA (siRNA), or locked nucleic acid, and wherein the exogenous nucleic acid binds to a target RNA and modulates gene expression of the target RNA, thereby treating Coronavirus disease 2019 (COVID-19) in the subject.

This disclosure relates to the modified 3' region elements that confer improved therapeutic properties including, but not limited to, increased stability, increased expression, and / or a reduced innate immune response when introduced into a population of cells, even in the absence of a 5'-cap. The disclosure also provides polynucleotides (e.g., RNA molecules) containing modified 3' regions. In some embodiments of the disclosure, the modified 3' regions contain modified adenosines (e.g., 2'-0-methoxy- adenosine, 2'-fluoro-adenosine, or locked nucleic acid (LNA)-adenosine) and / or a modified thymidine (e.g., inverted deoxythymidine). In some embodiments of the disclosure, the modified 3' region may contain one or more modified internucleoside linkages (e.g., a phosphorothioate linkage) or modified nucleobase (e.g., N1-methyl-pseudouridine). The polynucleotides of the disclosure may be used to express a desired polypeptide in a subject, such as a subject having a disease or condition associated with a deficiency in the corresponding endogenous polypeptide.

The present application relates to the field of biotechnology and molecular detection, in particular to a plant and pathogenic bacteria interaction gene expression analysis method based on UMI tag, comprising: obtaining interaction sample total nucleic acid extract; and constructing a molecular hybridization system with host-specific chimeric blocking probes containing locked nucleic acid modification and 3' end dideoxy modification, and reverse transcription primers; through temperature rising and denaturation and temperature reduction and annealing, the blocking probes are combined with host high abundance nucleic acid to form a hybridization blocking complex, and the primers are combined with pathogenic bacteria nucleic acid; template switching oligonucleotides with unique molecular identifier sequences and reverse transcriptase are added, the host extension is blocked, and UMI is introduced into the pathogenic bacteria complementary chain, and an interaction expression library is generated; after sequencing, the pathogenic bacteria effective reads and the host invalid reads are counted, and the effective interaction signal signal-to-noise ratio is output; the present application can preferentially capture pathogenic bacteria transcripts in the high background of host RNA.

Disclosed herein include systems, methods, compositions, and kits for prevention of antibody oligonucleotide extension. There are provided, in some embodiments, blocking oligonucleotides. In some embodiments, the blocking oligonucleotide comprises a binding region capable of hybridizing barcoding oligonucleotides to form a protected duplex. The blocking oligonucleotide can be a locked nucleic acid (LNA), a peptide nucleic acid (PNA), a DNA, an LNA / PNA chimera, an LNA / DNA chimera, or a PNA / DNA chimera. In some embodiments, the 3′ end of a blocking oligonucleotide is incapable of being extended by a reverse transcriptase or a polymerase. In some embodiments, the blocking oligonucleotide of a protected duplex is incapable of being removed by the strand displacement activity of a reverse transcriptase or a polymerase.

The invention provides a composition, a kit and a method for detecting African swine fever virus, and relates to the technical field of gene detection. The primer group and the probe provided by the invention are used for detecting the African swine fever virus, the probe is a ZIPNA probe, the ZIPNA probe comprises a 5 '-terminal fluorophore, a 3'-terminal quenching group and a spermine group, and the ZIPNA probe can provide higher detection specificity because non-specific binding and background signals can be more effectively reduced. The positive ions coupled with the ZIPNA probe can neutralize negative charges of target nucleic acid and reduce electrostatic repulsion, and meanwhile, the ZIPNA probe coupled with a certain number of positive ion groups can reach a preset Tm value. The ZIPNA probe can effectively replace a small groove conjugate and locked nucleic acid, and can be used in various application fields based on nucleic acid detection. The ZIPNA probe has the advantages of being high in detection fluorescence increment, high in detection specificity, good in repeatability, high in sensitivity and the like, and practical application is facilitated.

The disclosure herein includes methods, compositions, and kits for detecting a target nucleic acid sequence in a sample. The method may include generating an oligonucleotide (SGO) using a signal capable of hybridizing to a nucleic acid amplification product. The SGO may comprise a 5'subdomain, a 3 'subdomain, and a ring domain between the 5' subdomain and the 3 'subdomain. Intramolecular nucleotide base pairing between the 5'subdomain and the 3 'subdomain may be capable of forming a paired stem domain. The SGO may comprise one or more locked nucleic acid (LNA) nucleotides in the loop domain, the 5 '-subdomain, and / or the 3'-subdomain SGO may comprise a 5 '-terminal domain located 5' of the 5 '-subdomain In some embodiments, the 5'-terminal domain is not capable of hybridizing to the 3 '-terminal of the nucleic acid amplification product.

Disclosed herein include systems, methods, compositions, and kits for preventing antibody oligonucleotide extension. In some embodiments, blocking oligonucleotides are provided. In some embodiments, the blocking oligonucleotide comprises a binding region capable of hybridizing to the barcoded oligonucleotide to form a protected duplex. The blocking oligonucleotide may be a locked nucleic acid (LNA), a peptide nucleic acid (PNA), a DNA, an LNA / PNA chimera, an LNA / DNA chimera, or a PNA / DNA chimera. In some embodiments, the 3'terminal of the blocking oligonucleotide cannot be extended by a reverse transcriptase or polymerase, In some embodiments, the blocking oligonucleotide of the protected duplex cannot be removed by the strand displacement activity of the reverse transcriptase or polymerase.

The invention discloses a mutant gene enrichment method, a mutant gene detection method and a mutant gene detection kit based on Cas9 specific cleavage and isonucleotide magnetic beads. The method comprises the following steps: firstly, extracting free DNA; then identifying and cutting a completely matched wild type sequence by using Cas9 protein and specific sgRNA, and reserving the mutant DNA due to base mismatch; cas9 treated DNA and streptavidin magnetic beads containing a biotinylated oligonucleotide probe are incubated, guanine and cytosine in the probe are substituted by isoguanine and isocytosine respectively, and adenine and thymine are modified by locked nucleic acid, so that high-stability homodromous pairing is realized, and mutation DNA is selectively enriched; and finally, carrying out qPCR or sequencing detection on the enriched DNA. The kit has the characteristics of simplicity and convenience in operation, high specificity and high sensitivity, can be used for detecting single-base-level mutation, and is suitable for liquid biopsy, tumor early screening and genetic disease mutation detection.

A cell processing platform and method for selective isolation and activation of target cells, such as T-cells, NK-cells, and hematopoietic stem cells, from a biological sample. The platform employs aptamer-functionalized microbubbles, which selectively bind to target cell surface antigens to create buoyant target cell-microbubble complexes. A centrifugation module separates buoyant target cells from non-target cells by exploiting their distinct buoyancy properties. The system includes advanced aptamer designs, such as Locked Nucleic Acid (LNA) modifications, temperature-controlled folding buffers, and reversible binding mechanisms using complementary strands or enzymatic cleavage to release target cells. Additional features include optimized spacer configurations to enhance binding accessibility and aptamer stability for high specificity and yield. This platform also enables selective removal of senescent cells using aptamer-functionalized microbubbles, facilitating continuous processing and supporting applications in cell therapy, research, and regenerative medicine.

To provide a C6'-substituted locked nucleic acid-modified cap analog and a method of use thereof.SOLUTION: The present disclosure provides a compound represented by a formula (I), or a stereoisomer, a pharmaceutically acceptable salt or a solvate thereof. In the formula, X is O, S, or the like; R1 to R6 are each H, substituted / unsubstituted C1-C6 alkyl, C2-C6 alkenyl, or the like; Y1a to Y1f are each O, S, CH2, CCl2, or the like; Y2a to Y2d are each =O or =S; Y3a to Y3d are each OH or SH; and B1 and B2 are each a natural / modified pyrimidine nucleotide base or natural / modified purine nucleotide base.SELECTED DRAWING: Figure 4

The invention relates to a primer and a locked nucleic acid probe set for real-time fluorescent quantitative PCR detection of two goose parvovirus virulent and attenuated strains, the goose parvovirus virulent and attenuated strains comprise MDGPV virulent and attenuated viruses and SBDSV virulent and attenuated viruses, and the sequences of the primer and the locked nucleic acid probe set are respectively shown as SEQ ID NO.1-8. The LNA-TaqMan real-time fluorescent quantitative PCR detection method established by utilizing the primers and the LNA-TaqMan probe group can be used for simultaneously detecting strong and weak goose parvovirus strains (strong and weak muscovy duck gosling plague virus and / or strong and weak duck short beak and dwarf syndrome virus), identifying, diagnosing and accurately quantifying, and is high in sensitivity, strong in specificity and good in repeatability.

The invention relates to the technical field of microbiological detection, in particular to an LNA-qPCR accurate detection method for bifidobacterium longum subsp.infantis CICC 6069, a specific primer, a probe and a kit for the method and application of the specific primer, the probe and the kit. The invention provides application of a nucleotide sequence as shown in SEQ ID NO: 1 in precise identification and quantitative detection of a bifidobacterium longum subsp.infantis CICC 6069 strain. The method comprises the following steps: constructing a bifidobacterium longum generic genome, excavating a specific SNP (Single Nucleotide Polymorphism) site of CICC 6069, designing a locked nucleic acid (LNA) modified specific probe SEQ ID NO: 14 and primers SEQ ID NO: 4 and SEQ ID NO: 5, and combining a real-time fluorescent quantitative PCR (Polymerase Chain Reaction) technology, so that the rapid and accurate quantitative detection of the strain on the strain level is realized. The method has the advantages of strong specificity, high sensitivity (the quantitation limit is about 500 CFU / mL), short detection period and the like, and is suitable for quality control and quantitative analysis of samples such as foods, daily chemicals, complex microbial inoculants and the like containing CICC 6069.

To provide effective nucleotide or chemical motifs for dsRNA molecules, which are advantageous for inhibition of target gene expression, while having reduced off-target gene silencing effects; and RNAi compositions suitable for therapeutic use.SOLUTION: A double-stranded RNA (dsRNA) molecule capable of inhibiting expression of a target gene comprises a sense strand and an antisense strand, each strand having 14 to 40 nucleotides, where the antisense strand has sufficient complementarity to a target sequence to mediate RNA interference, where the antisense strand comprises at least one thermally destabilizing modification of a duplex within first 9 nucleotide positions of the 5' region or a precursor thereof, where the sense strand comprises an ASGPR ligand, and where the destabilizing modification is selected from a modified unlocked nucleic acid (mUNA) and a glycol nucleic acid (GNA).SELECTED DRAWING: None

This invention relates to the field of microbial detection technology, specifically to a precise LNA-qPCR detection method for *Bifidobacterium longum* subsp. *infantii* CICC 6069, as well as specific primers, probes, kits, and their applications for this method. This invention proposes the application of the nucleotide sequence shown in SEQ ID NO:1 in the precise identification and quantitative detection of *Bifidobacterium longum* subsp. *infantii* CICC 6069. By constructing a pan-genome of *Bifidobacterium longum*, mining SNP sites unique to CICC 6069, and designing specific probes SEQ ID NO:14 and primers SEQ ID NO:4 and SEQ ID NO:5 containing locked nucleic acid (LNA) modification, combined with real-time quantitative PCR technology, rapid and precise quantitative detection of this strain at the in vitro level is achieved. This method has advantages such as high specificity, high sensitivity (limit of quantification approximately 500 CFU / mL), and short detection cycle, and is suitable for quality control and quantitative analysis of samples containing CICC 6069, including food, daily chemical, and compound microbial agents.

The application discloses a method for detecting thyroid cancer driving gene mutations based on multiplex fluorescence PCR, and belongs to the technical field of molecular biology detection. The method simultaneously detects nine mutation sites, namely BRAF V600E, NRAS Q61R, NRAS Q61K, HRAS Q61R, HRAS Q61K, KRAS G12V, KRAS G12D, TERT C228T and TERT C250T in a single tube reaction. The method enhances the distinguishing ability of wild type and mutant types by using a locked nucleic acid modified allele-specific primer, suppresses wild type amplification by using a peptide nucleic acid clamp, introduces a competitive internal reference probe to correct amplification efficiency difference, and reduces the sample usage to one fifth of that of a traditional multi-tube scheme. The mutation detection lower limit reaches 0.5% allele frequency, and the total time consumption is not more than 2.5 h.

The present invention discloses a nucleic acid probe combination for detecting rifampicin resistance gene mutation of Mycobacterium tuberculosis, comprising: a plurality of probes for a nucleic acid amplification reaction, each of the plurality of probes being completely complementary to a corresponding one of a plurality of mutant sequences in a drug resistance determining region of an rpoB gene, and at least one of the plurality of probes has a locked nucleic acid (LNA) modification.

The invention discloses a double-end modified probe for detecting single nucleotide polymorphism as well as a design method and application of the double-end modified probe, and through the synergistic effect of thio-modification and locked nucleic acid modification and in combination with SNP sites and 3 '-end artificial mutation design, the distinction degree and detection sensitivity of the probe to wild types and mutant types are improved. Experiments show that the probe can detect a mutation proportion as low as 0.1%, the melting temperature difference between a wild type and a mutant type is greater than 4 DEG C, and the probe is suitable for multiple detection of drug-resistant genes or strain identification of pathogenic microorganisms such as mycobacterium tuberculosis and the like.

An oligonucleotide probe for detecting a genomic alteration includes a first complementary region, a second complementary region, and a semi-complementary region. The first complementary region includes at least three nucleotides that are completely complementary to a mutant allele of a portion of a genomic sequence, the includes at least three nucleotides that are completely complementary to the mutant allele and to the wild type allele, and the semi-complementary region is adjacently flanked by the first complementary region and the second complementary region. The nucleotides of the semi-complementary region are completely complementary to the mutant allele and are not completely complementary to the wild type allele, the first complementary region comprises a first locked nucleic acid adjacent to the semi-complementary region, the second complementary region comprises a second locked nucleic acid adjacent to the semi-complementary region, and the semi-complementary region comprises a third locked nucleic acid.

The present invention relates to RNA molecules, and in particular, but not exclusively, to unlocking nucleotides (UNAs) and unlocking nucleic acid (UNA)-RNA molecules themselves, as well as to methods of making UNA-RNA molecules, and to methods of translating UNA-RNA molecules into proteins. The present invention relates to an improved method of forming UNA-RNA by in vitro transcription, and to the resulting UNA-RNA molecules. Furthermore, the present invention relates to a novel method for enhancing the expression and / or translation of RNA, i.e. Protein expression, as well as a method for improving the stability of RNA molecules. The invention also relates to reducing the activation of innate perception, interferon production, and / or degradation of RNA molecules in a host. The invention also relates to the use of said UNA-RNA molecules in vaccines and other therapeutic pharmaceutical compositions, as well as their use in immunization and therapy, such as RNAi, gene therapy, gene editing and protein substitution. Furthermore, the present invention relates to a novel method of enhancing RNA translation, i.e. Protein expression.

The invention belongs to the technical field of biological detection, and particularly relates to a DNA polymerase activity detection method, a detection kit and application thereof. The method is based on a qPCR technology, and efficient and accurate detection of DNA polymerase activity is realized by designing a special reaction system and optimizing an experimental process. In the incubation stage, an oligonucleotide fragment which is annealed in advance and contains mismatched basic groups is used as a primer and a template of DNA polymerase to be detected to carry out polymerization reaction; in a detection stage, a polymerization reaction product is quantified by virtue of a qPCR technology through a locked nucleic acid enhanced TaqMan probe competitive binding strategy. The method is simple and convenient to operate, good in safety, high in sensitivity and wide in linear range, and can be applied to the scenes of activity detection of different types of DNA polymerase, enzyme preparation screening, hot start polymerase performance evaluation, polymerase activity quantification in complex samples and the like, so that the method has good practical application value.

The present application relates to the technical field of biological detection, and a double-path apple codling moth identification system integrating RPA-LFS rapid screening and TaqMan qPCR real-time quantification, a locked nucleic acid probe primer set, a kit and a method thereof. COI Through gene screening and primer screening, the present application designs recombinase-mediated isothermal amplification (RPA) and TaqMan qPCR primers and probes for the barcode of the cytochrome oxidase subunit I (COI) gene of apple codling moth, and establishes an RPA-LFS method in combination with immunolateral flow. The RPA-LFS method can complete amplification and visual judgment within 20 minutes at 39 DEG C, and is suitable for on-site rapid detection. The TaqMan qPCR method can realize high-sensitivity quantitative analysis under the condition of crude extract. Through double-path complementary design, the consistency of the results of on-site screening and laboratory confirmation detection is realized, and the method is simple to operate and suitable for pest quarantine at ports, epidemic monitoring in agricultural production areas and quarantine at storage and transportation links.