63 results about "Target enrichment" patented technology

The invention discloses a detection system and method for identifying mycobacteria and drug resistance genes of the mycobacteria based on nanopore sequencing, and relates to the field of biological medicine, the detection system comprises a specific targeted enrichment module, a nanopore sequencing module and a biological information analysis module; the specific targeted enrichment module comprises a probe combination, and the probe combination covers a mycobacterium tuberculosis complex conservative identification gene, species-specific genes of common nontuberculous mycobacteria and mycobacterium leprosy, and full-length or partial sequences of drug resistance related genes in a targeted manner; by utilizing the characteristics of nanopore length reading length and real-time sequencing and a tuberculosis specific targeted enrichment strategy, accurate identification of a mycobacterium tuberculosis complex group, synchronous typing of 42 mycobacteria, analysis of 24 drug-resistant genes, and efficient detection of structural variation and low-abundance heterogeneity drug-resistant mutation are realized, the detection period is shortened, the detection cost is reduced, and the detection efficiency is improved. And a comprehensive and reliable technical basis is provided for accurate diagnosis and treatment of mycobacterium infection.

The invention discloses a sequencing method of an immune repertoire, which comprises the following steps: 1) constructing a cDNA library of a sample, the cDNA in the cDNA library being connected with the position information of the cDNA on the sample; 2) enriching cDNA of TCR and / or BCR genes in the cDNA library in a targeted manner to obtain an immune repertoire of the sample; and (3) sequencing the immune repertoire. According to the method disclosed by the invention, targeted enrichment and high-throughput sequencing of TCR and / or BCR sequences are innovatively realized, and gene expression information and spatial position information of each cell in a space are detected at the same time.

The invention relates to a drug-resistant gene targeted enrichment method using dCas9 and application of the drug-resistant gene targeted enrichment method, and belongs to the technical field of nucleic acid enrichment. The drug-resistant gene targeted enrichment method comprises a CRISPR-dCas9 system for targeting a target gene and a target capture magnetic bead matched with the CRISPR-dCas9 system; the CRISPR-dCas9 system comprises a target DNA (Deoxyribose Nucleic Acid), a dCas9 protein and a target nucleic acid targeted sgRNA (Single Guide Ribonucleic Acid), and the targeted capture magnetic bead is a His tag protein purified agarose magnetic bead. According to the method, a CRISPR-dCas9 system and His tag protein are integrated to purify agarose magnetic beads, and dCas9 protein with a His tag is combined with the magnetic beads to construct a novel enrichment scheme; his magnetic beads are combined with dCas9 with a His tag under the condition that protein denaturation is not achieved, and drug-resistant gene targeted enrichment is achieved; the whole scheme is easy to operate, mild in condition, low in cost and suitable for common laboratories and on-site rapid detection to improve sensitivity. According to the method, the target gene is highly specifically recognized through sgRNA, magnetic bead capture is combined, and the method has the advantages of being efficient, specific and easy to operate and has application value in the fields of drug-resistant gene screening and diagnosis, third-generation sequencing and the like.

The invention provides methods for simultaneously enriching multiple target regions of interest in one reaction volume, from cell-free DNA isolated from a blood or plasma sample, followed by high-thought sequencing and sequence read analysis. The invention also provides library of target-specific oligonucleotide primers or probes for the multiplexed target enrichment.

This invention relates to the field of bioinformatics processing and discloses a method and system for mining the structure-activity relationship (SMR) of porcine neurotrophic peptides based on sequence features. The method includes constructing an original sample index table and fusing multi-source production data, extracting peptide sequence features to generate a sequence feature matrix, constructing a sequence-process joint graph containing peptide nodes and process state nodes, training a structure-activity relationship graph neural network to mine SMR relationships, and deriving a process control decision table based on a process response sample set generated by the network, thereby achieving online optimization of the porcine neurotrophic peptide preparation process. This invention solves the problem of SMR mining caused by the separation of process parameters, sequence information, and activity data, achieving accurate characterization of the synergistic effect of sequence and process and reverse optimization of process parameters, thus improving the targeted enrichment efficiency and bioactivity retention level of target neurotrophic peptides.

The present disclosure provides, among other things, a way to amplify and sequence target sequences in a low-input sample. In some embodiments, the method comprises ligating a double-stranded adaptor onto a population of fragments to produce tagged fragments, and linearly amplifying the tagged fragments.

This invention discloses a mitochondrial-targeted nanomaterial for senescent cells and its application in anti-aging. The material is a nucleic acid aptamer-tetrahedral DNA, mainly assembled from four single-stranded DNA sequences, SEQ ID Nos. 1-4. Preparation involves first hybridizing the four single-stranded DNA sequences, then concentrating them to obtain aptamer-modified tetrahedra, which are then coupled with tannic acid (TA) via a Mann reaction in the presence of formaldehyde to obtain the Apt-tFNAs-TA complex. Finally, it is incubated with excess catechins to obtain the final delivery material, Apt-tFNAs@catechin-TA. This invention's material exhibits good biocompatibility and skin tissue compatibility, significantly improving uptake efficiency and mitochondrial localization ability in senescent cells. It can effectively deliver polyphenolic compounds such as catechins and protocatechuic acid, improving mitochondrial function and addressing the technical problems of low delivery efficiency of active ingredients and insufficient targeted enrichment in senescent cells.

Provided are a humanized / fully human HER2 antibody or an antigen-binding fragment thereof, and an antibody-radionuclide conjugate using same as a targeting group for radioactive labeling, showing a tumor-targeting enrichment ability.

The application discloses a kind of enrofloxacin rapid detection methods based on fluorescent covalent organic framework magnetic beads, belong to food safety detection field.The method uses the fluorescent covalent organic framework magnetic bead of coupling enrofloxacin monoclonal antibody, realizes detection in combination with immunochromatography test strip.Fluorescent covalent organic framework magnetic bead realizes target enrichment separation with inner core, shell fluorescent covalent organic framework material high-efficiency coupling antibody and exports fluorescence signal;Test strip is equipped with detection line (coated enrofloxacin artificial antigen) and quality control line (coated secondary antibody).By comparing the fluorescence intensity of detection line and quality control line, trace enrofloxacin in complex matrix can be qualitatively and quantitatively analyzed.The method is simple, rapid and sensitive, without large instrument, can be detected on site.

The invention belongs to the technical field of microbiological detection and molecular diagnosis, and discloses a primer group, a kit and a detection method for detecting salmonella enteritidis, the kit comprises an immunocapture component, an isothermal amplification component, a CRISPR / Cas12a detection component and auxiliary components; the isothermal amplification component is any one of an RPA (recombinase polymerase amplification) component or an LAMP (loop-mediated isothermal amplification) component, the RPA component comprises a specific primer group targeting 86th to 212th nucleotides in a salmonella enteritidis hilA gene coding region, and the LAMP component comprises a specific primer group targeting 58th to 250th nucleotides in the salmonella enteritidis hilA gene coding region. The primer group, the kit and the detection method for detecting the salmonella enteritidis successfully integrate three key technologies of RAP and LAMP rapid isothermal amplification, CRISPR / Cas12a high-specificity recognition and trans-cleavage and immunomagnetic bead targeted enrichment, and creatively provide the primer group, the kit and the detection method for detecting the salmonella enteritidis.

The invention discloses a multi-target enriched library construction method for simultaneously detecting DNA (deoxyribonucleic acid) and RNA (ribonucleic acid) type pathogens, which is characterized in that after total nucleic acid of a sample is extracted, DNA and RNA libraries are simultaneously constructed in a single-tube reaction, and the proportion of the pathogens in the library is enriched by capturing a probe, so that the sensitivity of pathogen detection is improved, the sequencing data volume is reduced, the library construction steps are reduced, and the detection efficiency is improved. The method has the advantages that the method is simple and convenient to operate, the condition that samples are misplaced possibly due to the fact that RNA is firstly processed into two-chain cDNA and then mixed into DNA to form cDNA / DNA compounds is avoided, only one-tube extraction can be carried out, and the method has high application value on samples such as neonatal blood and child cerebrospinal fluid under the condition that the sample size is small.

The present disclosure provides for methods of preparing nucleic acid libraries for sequencing. In particular, the present disclosure is directed to a workflow which employs a first round of amplification utilizing a universal polymerase chain reaction approach instead of a platform specific indexed polymerase chain reaction approach. Applicant has unexpectedly discovered that a workflow employing a first round of amplification which utilizes a universal polymerase chain reaction approach increases duplex recovery and / or genomic equivalent recovery as compared with a workflow employing a first round of amplification utilizing a platform specific indexed polymerase chain reaction.

Disclosed are methods for preparing a non-naturally occurring composition, comprising: extracting cell-free DNA from a plasma fraction of a blood sample of a pregnant person, wherein the pregnant person is a heterozygous carrier of at least one pathogenic variant in at least one target gene associated with autosomal recessive disorders, wherein the extracted cell-free DNA comprises a mixture of maternal cell-free DNA and fetal cell-free DNA; performing targeted enrichment on the extracted cell-free DNA or DNA derived therefrom to enrich a plurality of target variant loci in a plurality of target genes and generating enriched DNA, wherein the target variant loci comprise the at least one pathogenic variant; performing high-throughput sequencing on the enriched DNA or DNA derived thereof and generating sequence reads, and determining fetal genotypes of one or more of the target genes from the sequence reads.

The invention discloses a multiple myeloma sequencing method based on a targeted capture three-generation sequencing technology, a kit and application, and relates to the technical field of gene detection, and the method comprises the steps of sample treatment, targeted probe group design and targeted enrichment capture, three-generation sequencing library preparation, bioinformatics analysis and the like to identify and analyze three types of variations. According to the method, the defects that an existing layered system lacks an accurate genomics basis, and FISH and NGS technologies and combination strategies are low in flux, incomplete in coverage, incapable of synchronously detecting various variations, high in cost and complex in process are overcome, comprehensive information of three key variation types of multiple myeloma SNV, SV and CNV can be obtained at the same time through a single experiment and one-time sequencing, and the method is suitable for popularization and application. The detection efficiency is improved, and sample consumption, operation time and cost are reduced; all hot spot areas and translocation breakpoint areas of the most important clinical related genes of the MM are covered, and a gene variation map which is more comprehensive than that of a standard FISH Panel is provided.

This invention discloses a targeted sequencing method for multiple pathogenic microorganisms, belonging to the field of biotechnology. The targeted sequencing method for multiple pathogenic microorganisms includes the following steps: sample pretreatment; targeted enrichment; PCR amplification; high-throughput sequencing; and bioinformatics analysis. This invention solves the problems of existing technologies, such as inability to accurately type pathogens, detect drug resistance genes, and analyze virulence genes, as well as poor targeting and slow analysis speed. This invention provides high-purity target nucleic acids for PCR amplification through a triple targeted enrichment system; the anti-inhibition multiplex PCR system ensures efficient amplification of target nucleic acids in complex samples; high-throughput sequencing enables rapid sequencing of target nucleic acids; and the bioinformatics analysis module enables simultaneous completion of pathogen typing, drug resistance gene detection, and virulence gene analysis, significantly improving overall detection efficiency and meeting the clinical needs for rapid, accurate, and comprehensive pathogen detection.

The application discloses a target gene for detecting pea foot rot, a primer set and a detection kit, and belongs to the technical field of plant quarantine. The application innovatively develops a rapid detection technology system based on specific molecular markers in view of the core pain points of long detection period (traditional method >= 20 days) and difficult identification of close species (such as D. pinodes and A. pisi) in the quarantine identification of Didymella pinodella. By screening a single-copy gene sequence unique to D. pinodella and combining PCR technology, a whole-process system of "direct sample detection-target enrichment-signal amplification" is established, and the pathogen detection can be completed within 8 hours, the efficiency is improved by 96% compared with the traditional method, the efficiency is greatly improved, and the close species can be effectively distinguished, the customs clearance efficiency is greatly improved, and technical support is provided for the detection of pea foot rot and the breeding of pea disease resistance.

The invention discloses a kit for detecting circulating tumor DNA (deoxyribonucleic acid) of portal vein and branch vein blood of portal vein. The kit comprises: a nucleic acid extraction assembly comprising an optimized lysate and selectively bound magnetic beads; the library construction component comprises a terminal repair / tailing fusion enzyme and a Y-shaped linker with a molecular identifier; the targeted enrichment assembly comprises a specific primer group and a probe group and is used for enriching related gene targets of the abdominal organ tumors; and necessary reaction components. Through an optimized nucleic acid extraction method, an efficient library construction system and a precise targeted enrichment technology, high-sensitivity detection of low-abundance ctDNA in portal venous blood is achieved, and the lower detection limit reaches 0.1% of mutation allele frequency. The kit is particularly suitable for early diagnosis, curative effect monitoring and prognosis evaluation of abdominal organ tumors such as intestinal cancer, liver cancer and pancreatic cancer.

The invention provides a genetic variation and chimera detection method, system and kit. The method comprises the following steps: (a) extracting high molecular weight DNA from a biological sample; (b) labeling the high molecular weight DNA by using a linker containing a UMI sequence to form a UMI labeled DNA library; (c) performing targeted enrichment on the DNA library marked by the UMI by using a probe aiming at a target gene locus to obtain an enriched long-fragment DNA library; (d) performing long read length sequencing on the enriched long fragment DNA library to obtain sequencing data; and (e) carrying out bioinformatics analysis on the sequencing data, wherein the bioinformatics analysis comprises error correction and variation detection based on a UMI sequence. The method is used for simultaneously detecting structural variation and low-frequency chimeric variation in a target genome region.

To provide a method for enrichment of at least one target nucleic acid in a library of nucleic acids.SOLUTION: A first oligonucleotide is hybridized to a target nucleic acid in library of nucleic acids having first and second adapters. The hybridized first oligonucleotide is extended with a first polymerase, thereby producing a first primer extension complex including the target nucleic acid and the extended first oligonucleotide. The first primer extension complex is captured, enriched relative to the library of nucleic acids, and a second oligonucleotide is hybridized to the target nucleic acid. The hybridized second oligonucleotide is extended with a second polymerase, thereby producing a second primer extension complex including the target nucleic acid and the extended second oligonucleotide, and further liberating the extended first oligonucleotide from the first primer extension complex.SELECTED DRAWING: Figure 1

The application provides a circulating tumor DNA-based mutant gene detection method and application thereof, and relates to the technical field of biological detection.The application provides a circulating tumor DNA-based mutant gene detection method, adopts a customized panel technical route, adopts a single-stranded library construction method, preliminarily amplifies free DNA, and then performs target enrichment based on UMI multiplex PCR, so that the detection sensitivity and specificity are effectively improved.It is proved through tests that the method can simultaneously analyze multiple gene mutation results of the same patient, has a sensitivity of more than 70% and a specificity of more than 90% on the mutation allele ratio (AF), and has a good application prospect for MRD monitoring.

The application discloses a kind of magnetic nanomaterials for targeting enrichment protein and protein sample processing method using the magnetic nanomaterials, and nanomaterial has shell-core structure, inner core is ferroferric oxide magnetic nanoparticle, i.e.magnetic core, outer shell is mesoporous titanium dioxide, mesoporous titanium dioxide is coated on the surface of magnetic core, the mesoporous titanium dioxide has multiple nanopores, can be modified by molecular glue, can specifically bind target protein, when processing protein sample, magnetic nanomaterial is introduced into the complex biological sample containing protein, nucleic acid and salt, and magnetic nanomaterial selectively binds target protein, both can separate the required target protein, obtain peptide segment after digestion elution, and be used for proteomics data acquisition and analysis, the material and method of the application can improve protein fixation recovery efficiency before proteomics determination, save sample, save time and reduce cost.

The invention provides methods for simultaneously enriching multiple target regions of interest in one reaction volume, from cell-free DNA isolated from a blood or plasma sample, followed by high-thought sequencing and sequence read analysis. The invention also provides library of target-specific oligonucletotide primers or probes for the multiplexed target enrichment.

The present disclosure is directed to compositions, kits, and methods of target enrichment by unidirectional primer extension, whereby the compositions, kits, and methods utilize both poison primers and target capture primers.

The invention discloses a PCR (Polymerase Chain Reaction) primer group for detecting bovine nodule skin disease virus based on multiple amplicon targeted enrichment and nanopore real-time sequencing as well as application and a detection product thereof, and relates to the technical field of biological detection. The PCR primer group comprises primers of which the nucleotide sequences are as shown in SEQ ID NO. 1-90. According to the invention, a multiple PCR primer group is designed based on a bovine nodular skin disease virus genome sequence, and a detection method for combining multiple amplicon targeted enrichment and nanopore real-time sequencing aiming at bovine nodular skin disease pathogens and a corresponding detection product are established according to the multiple PCR primer group. By adopting the detection product provided by the invention, nanopore real-time sequencing can be carried out by aiming at the specific amplified fragment of the primer group of the bovine nodular dermatosis pathogen, so that on-site accurate and rapid detection of the bovine nodular dermatosis pathogen is realized, pathogen detection is carried out from a sequence level, and three viruses, namely LSDV, SPPV and GTPV, are accurately distinguished.

The invention relates to the technical field of electron microscope sample preparation, and discloses a charge density enhanced hydrophobin HFBI mutant mesh film for a cryoelectron microscope and a preparation method and application thereof. The invention provides a surface charge enhanced HFBI mutant. Targeted enrichment and orientation control on different samples are enhanced by designing charge distribution on an electrostatic interface. According to the hydrophobic protein HFBI mutant carrier network disclosed by the invention, a plurality of key bottlenecks in preparation of a cryoelectron microscope sample are systematically overcome by virtue of a synergistic effect of three aspects of resolution improvement, orientation regulation and enrichment enhancement, and a powerful tool is provided for analyzing a high-resolution three-dimensional structure of biological macromolecules.

This invention discloses a method for preparing DNA samples for SNP chip detection, comprising the following steps: obtaining at least one library sequence, wherein the library sequence includes a capture sequence complementary to the target fragment of the DNA sample to be tested; amplifying the library sequence and then digesting it with enzymes to obtain a single-stranded library sequence; mixing the amplification product of the DNA sample to be tested with the single-stranded library sequence, denaturing and annealing; and digesting the product with Exonuclease I. The DNA sample prepared by this invention is more purified, with targeted enrichment of the target fragment and removal of a large number of non-target fragments. When applied to SNP chip detection, it improves the hybridization efficiency with the SNP chip, thereby increasing the detection signal value and accuracy.

The invention discloses a chlamydia psittaci liquid-phase probe targeted capturing and library building method and a kit thereof, and belongs to the technical field of virus detection, a specific probe is designed to enrich pathogen DNA in a targeted manner, a sample treatment process is optimized, targeted enrichment and high-throughput sequencing are combined, and the method has the advantages of wide coverage, good flexibility, high sensitivity, high accuracy and the like. The method has the characteristics of high sensitivity, high specificity and the like, and can be used for quantitative, qualitative and deep analysis and other multi-dimensional information output. By designing the specific probe and optimizing the sample treatment and library building process, the sensitivity, specificity and efficiency of chlamydia psittaci detection are remarkably improved, and the blank in the aspects of low-load detection, whole-genome analysis and high-throughput adaptation in the prior art is filled; the method has important application value in the fields of clinical diagnosis, poultry epidemic disease prevention and control and public health safety.

This invention provides a method for targeted enrichment of target sequences in a spatial transcriptome cDNA library. The method includes: using a hybridization capture probe set to hybridize and extend a target nucleic acid sequence or its complementary sequence on the target sequence in the spatial transcriptome cDNA library to obtain an extension product; wherein the hybridization capture probe set includes two or more probes, and any two probes in the hybridization capture probe set do not overlap in the hybridization region of the target sequence; the extension direction of any probe in the hybridization capture probe set is from the target nucleic acid sequence or its complementary sequence to the spatial sequence or its complementary sequence; the extension product includes: the spatial sequence or its complementary sequence, and part or all of the target nucleic acid sequence or its complementary sequence; the extension product is isolated to obtain an enriched library. This method solves the problem in existing technologies that only unbiased sequencing can be performed on spatial transcriptome libraries, and targeted sequencing cannot be achieved, making it applicable to the field of spatial transcriptome library enrichment.

Embodiments include methods and systems for quantifying the efficiency of enriching for a target nucleic acid in a nucleic acid sample. Some embodiments include methods and electronic systems for implementing such methods. Some embodiments include adding control polynucleotides to a nucleic acid sample; hybridizing a probe set to the control polynucleotides and nucleic acid sample; amplifying or separating any hybridized nucleic acids, thereby providing an enriched library; sequencing the enriched library; and quantifying the efficiency of the enrichment for the targeted regions based on the proportion of targeted control sequence reads to untargeted control sequence reads.

The present disclosure provides a method of enriching a variant nucleic acid molecule from a population of nucleic acid molecules, said method comprising contacting a population of nucleic acid molecules with a primer in a reaction mixture, incubation of the reaction mixture for hybridization of the primer with a nucleic acid molecule, incubation of the reaction mixture for primer extension though the action of a polymerase, contacting the population of nucleic acid molecules in the reaction mixture with a nuclease, purifying duplex nucleic acid molecules comprising the variant nucleic acid molecule. The present disclosure also provides a method of designing a primer.