100 results about "Locked nucleic acid" patented technology

The invention relates to the technical field of molecular biology, in particular to a specific probe, a primer probe combination for detecting mycoplasma pneumoniae drug-resistant mutation and bordetella pertussis drug-resistant mutation and a kit. The primer probe combination and the kit provided by the invention can distinguish mutation at sites 2063 and 2064 of mycoplasma pneumoniae non-mutation sensitive strains and 23s rRNA genes as well as mutation at sites 2047 of bordetella pertussis non-mutation sensitive strains and 23s rRNA genes, and are relatively high in detection sensitivity; the sensitivity of a mycoplasma pneumoniae sensitive strain, a drug-resistant mutant strain and a bordetella pertussis sensitive strain can reach 25 copies / mL, high sensitivity and specificity are achieved by using locked nucleic acid modification at a specific position on a probe and specially optimized metal catalytic ions, and meanwhile, the reagent can be stably stored for 12 months at the temperature of 2-8 DEG C.

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 present invention relates to RNA molecules, and particularly, although not exclusively, to methods for preparing RNA molecules, and to methods for translating RNA molecules into protein. The invention extends to improved methods for forming RNA by in vitro transcription, and to the resultant RNA molecules. Furthermore, the invention relates to novel methods for enhancing the expression and / or translation of RNA, i.e. protein expression, and to methods for improving the stability of an RNA molecule. The invention also involves reducing the activation of innate sensing, interferon generation and / or degradation of an RNA molecule in a host. The invention also incorporates the use of the RNA molecules in vaccines and other therapeutic pharmaceutical compositions, and their use in immunisation and therapy, such as RNAi, gene therapy, gene editing and protein replacement. In particular, the invention relates to locked nucleotides and locked nucleic acid (LNA)-RNA molecules, and the use or incorporation of such locked nucleotides and LNA-RNA molecules in any of the above applications.

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 present application relates to the field of genetic engineering technology, and specifically, to a method for preparing a transcription reaction solution, a locked nucleic acid-modified RNA, and an application of a mutated T7 RNA polymerase. A transcription reaction solution for preparing locked nucleic acid-modified RNA comprises a DNA template, a mutated T7 RNA polymerase, LNA-NTPs, and a buffer; wherein the amino acid sequence of the mutated T7 RNA polymerase is shown in SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3, SEQ ID NO.4, SEQ ID NO.5, SEQ ID NO.6, or SEQ ID NO.7. The present application can significantly improve the transcription yield of locked nucleic acid-modified RNA by adding the above-mentioned mutated T7 RNA polymerase to the transcription reaction solution for synthesizing locked nucleic acid-modified RNA, and the operation is simple, which is conducive to industrial production.

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.

Interlocking nucleic acids for target hybridization are described. The interlocking nucleic acids include individual nucleic acids having respective capture regions that hybridize to the target nucleic acid and interlocking regions that bind to each other. In an embodiment, the interlocking nucleic acids are part of a probe set that specifically binds to a target nucleic acid to permit capture of the target nucleic acid. In an embodiment, the interlocking nucleic acids may be part of a primer used for amplification of the target nucleic acid.

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.

The present application relates to the aptamer specific to diethylstilbestrol and its application, and belongs to the technical field of biology.Through the capture-SELEX technology, three aptamer sequences R3, R7 and R9 with affinity and specificity to diethylstilbestrol are screened out.The best aptamer R7 is optimized by truncation, and finally a 27 nt long diethylstilbestrol aptamer R7-3-1 is obtained.On this basis, a higher performance diethylstilbestrol non-natural nucleotide aptamer sequence R7-3-1.6 is obtained by introducing locked nucleic acid modification, which significantly improves the environmental stability of the aptamer, and the affinity to diethylstilbestrol is improved compared with the original aptamer.Combined with the signal amplification ability of CRISPR-Cas12a, a fluorescence sensor is constructed to detect diethylstilbestrol.The present application provides a new recognition element and detection method with excellent performance for diethylstilbestrol detection.

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.