37 results about "Hybridization capture" patented technology

The present disclosure provides a method for detecting the methylation and mutation status of a DNA sample, specifically, a method for simultaneously detecting the methylation and mutation status of DNA molecules in a sample, the method comprising treating the sample with a methylation-sensitive restriction enzyme, performing hybrid capture on amplification products of the restriction enzyme-treated sample and a sample that has not been subjected to enzyme cleavage treatment with a probe set including a mutation capture probe and a methylation capture probe, and sequencing the captured products.

Provided are a method and a combination for one-tube simultaneous detection of tumor-specific gene mutation and methylation, and the use thereof. The method comprises searching a database to select mutation and methylation sites of a cancer driver gene and a cancer suppressor gene, synthesizing capture probes correspondingly paired with a target interval, performing hybrid capture on an amplification product of a sample treated with a restriction endonuclease by using a probe set comprising tumor-specific gene mutation and methylation capture probes, constructing a high-throughput sequencing library containing a target region, and performing high-throughput sequencing to obtain the sequence of a target fragment, so as to obtain the results of methylation, point mutations and indel variation of a cancer-associated gene. The provided method enables one-tube simultaneous detection of mutation and methylation states of a cancer-associated gene in one assay, requires a low content of a gene to be detected, can be used for methylation and mutation detection of a tumor-specific DNA, and has low detection costs and high efficiency.

The present application relates to the technical field of gene detection, in particular to a SNP molecular marker for detecting donor cfDNA of a pregnant organ transplant patient, a detection method and device. The SNP molecular marker for detecting donor cfDNA of a pregnant organ transplant patient is shown in Table 10. The present application uses free DNA in peripheral blood of a pregnant patient and genomic DNA in peripheral blood of a biological father of a fetus as detection objects, adopts liquid phase hybridization capture and high-throughput sequencing technology, uses thousands of SNP sites, obtains the genotype of the fetus through the genotype of the couple, eliminates the influence of cfDNA from the fetus on the content of cfDNA from the transplanted organ, accurately determines the concentration of cfDNA from the transplanted organ, has higher detection sensitivity, can be detected multiple times only by using peripheral blood of the patient, and achieves the purpose of long-term dynamic monitoring.

This application provides a method and system for infectious disease risk prediction based on a large language model and wastewater monitoring. The method includes: acquiring a multimodal time-series dataset of a target area in a first time period, wherein the multimodal time-series dataset includes wastewater epidemiological data sequences of the target disease, vaccination data sequences of the target disease, clinical monitoring data sequences of the target disease, environmental data sequences, and demographic data sequences obtained based on technologies such as multipathogen hybridization capture sequencing and PCR; converting the multimodal time-series dataset into a text time-series data sequence; inputting the text time-series data sequence into a preset risk prediction model to generate corresponding feature vectors, and predicting the trend of hospitalization rate changes of the target disease based on the feature vectors; and predicting the risk level of the target disease in the target area based on the trend of hospitalization rate changes and the wastewater epidemiological data sequence, thereby improving the accuracy of infectious disease area risk prediction.

The invention discloses a library construction method for non-HPV (human papillomavirus) related cervical tumor and precancerous lesion molecular subtypes thereof based on high-throughput sequencing, which comprises the following steps: ultrasonically breaking genome DNA (deoxyribonucleic acid) of a detected sample to obtain a DNA fragment, modifying the tail end, connecting a joint, and performing purification and pre-amplification reaction, namely performing hybridization reaction on a pre-amplification sequence by using a hybridization capture reagent, according to the method, a hybridization capture reagent is adopted, DNA fragments combined through non-specific capture are enriched and removed, then an amplification reaction is carried out, a library is obtained, next-generation sequencing is carried out, original data are subjected to bioinformatics analysis, variation information is obtained, and the hybridization capture reagent comprises specific primers with the sequences shown as SEQ ID NO: 1-175. According to the present invention, the integration of the non-HPV-related cervical tumor and precancerous lesion molecule detection is achieved, the detection can be completed in one experiment, and the sequencing depth is improved by specifically enriching the target region so as to improve the sensitivity of the non-HPV-related cervical tumor and precancerous lesion molecule detection.

The invention discloses a mutation back noise filtering algorithm based on next-generation sequencing data, relates to the technical field of high-throughput sequencing data analysis, and aims to solve the problems that the prior art depends on a baseline, and is poor in adaptability and high in false positive. The algorithm comprises the following steps: preprocessing sequencing data to obtain a pileup file, classifying sites according to a sequence context, calculating an error rate, constructing a layered back noise model to check mutation, qualitatively determining real mutation and back noise by combining a sample type, and evaluating sample quality. The method does not need to construct a base line in advance, adapt to hybrid capture and amplicon sequencing, can improve mutation detection specificity and repeated sample consistency, guarantees result reliability, and is suitable for scenes such as tumor gene detection.

The invention discloses a preciseness-adjustable probe hybridization capture method which is characterized in that a hybridization resolution enhancing probe is added into a system of a probe hybridization capture target area, a potential non-specific capture area is competitively combined, and the hybridization capture resolution of the probe to the target area is improved. The binding strength of the hybridization resolution enhanced probe and a potential non-specific capture region is higher than that of the probe and the potential non-specific capture region, and the binding strength of the hybridization resolution enhanced probe and a target region is far lower than that of the probe and the target region, so that the detection sensitivity of the target region is not influenced. The method realizes local fine control of hybridization capture preciseness of the nucleic acid target area, and simultaneously achieves high-sensitivity and high-specificity detection of the nucleic acid target area compared with a traditional method.

The invention belongs to the technical field of genotyping, and particularly relates to a Tan sheep 50K liquid phase chip based on targeted capture sequencing and application thereof, the Tan sheep 50K liquid phase chip is composed of independently packaged probe mixed liquid and a hybridization capture reagent, the independently packaged probe mixed liquid comprises a Tan sheep 50K probe, and site information of the Tan sheep 50K probe relates to 50000 SNP sites. The prepared Tan sheep 50K liquid chip is mixed with a Tan sheep high-throughput sequencing library, a target site is captured and amplified and purified, and after a product is subjected to high-throughput sequencing, a sequencing result is compared with a sheep reference genome, so that the genetic typing of the Tan sheep to be detected is obtained. The method is applicable to resource evaluation, genetic identification and genome selection of the Tan sheep variety, can break through key technical evaluation restricting genome selection breeding, lays a foundation for building a Tan sheep genome selection sharing platform, and accelerates the realization of the Tan sheep from phenotype selection to genome selection.

The invention belongs to the technical field of gene sequencing, and particularly relates to a library building kit and probe for detecting blood tumor related fusion genes. According to the library building kit, only a trace amount of total RNA is needed to be used for cDNA synthesis, the initial quantity of a template can be as low as 10 ng, so that a cDNA library is built, then the library is subjected to hybrid capture and enrichment by using an autonomously designed probe hematologic tumor fusion gene Panel (RA Panel), the time consumption is short, the efficiency is high, and simultaneous building and on-machine sequencing of 384 libraries can be realized. According to the rapid hybridization system of the library building kit, the shortest hybridization time can reach 30 min, the size of a finally obtained library fragment is about 200-400 bp, and the rapid hybridization system can be compatible with various Illumina sequencing platforms for sequencing. According to the invention, through a simple and easy-to-use solution of NGS sequencing while synthesis, rapid and accurate typing of fusion genes which may affect blood tumor diagnosis is realized.

The invention belongs to the technical field of library construction, and particularly relates to a DNA targeted library construction kit and application thereof. The kit comprises a hybridization capture reagent combination, the hybridization capture reagent combination comprises a hybridization reaction solution, and the hybridization reaction solution comprises a hybridization reaction solution 1 and a hybridization reaction solution 2; the hybridization reaction solution 1 is prepared from betaine, tetramethylammonium chloride, Mg < 2 + > and Triton X-100; the hybridization reaction liquid 2 is prepared from formamide and N-methyl pyrrolidone; hybridization of the probe and the library is facilitated, the capture efficiency is superior to that of an existing hybridization reaction solution, and Panel with different sizes and different hybridization durations can be compatible; a small panel can also achieve relatively high and relatively stable capture efficiency, the hybridization time is selectable within 1-20 h, fast and slow hybridization reagents are unified, and production and simultaneous operation of multiple products are facilitated.

The present application belongs to the field of medical detection, and specifically provides a detection method for solid tumor molecular residual lesions, which comprises tissue baseline detection, probe design, peripheral blood MRD detection and result analysis interpretation. Through the improvement of technical steps such as optimized pre-library construction and hybridization capture process, customized probe design and unique fusion gene probe design, the present application solves the problems of low detection sensitivity, low specificity and low accuracy in the prior art.

Provided herein are systems, kits, compositions and methods for sequencing library preparation and sequencing workflow (e.g., for the identification of mutations). In certain embodiments, provides herein systems and methods to identically barcode both strands of templates, and PCR-based enrichment of each strand that does not require hybridization capture.

The invention belongs to the technical field of molecular detection, and discloses a nucleic acid hybridization capture immunofluorescence detection kit as well as a use method and application thereof. In the kit provided by the invention, guanidine hydrochloride, guanidine isothiocyanate, triton X-100, NaCl, protease K and lauryl sodium sulfate are compounded according to a specific concentration to obtain the nucleic acid release liquid, so that pathogen nucleic acid in a clinical sample can be fully dissociated, and the nucleic acid release liquid can cooperate with a probe aiming at a specific target pathogen; the solution system can be used for optimizing the recognition and combination process of DNA and RNA, the specific recognition and combination capacity between the probe and specific pathogen nucleotide is improved, and under the condition that a common nucleic acid stabilizer, namely Tween, is not contained, the solution system can further strengthen differential structural epitopes existing in a DNA / RNA complex compared with DNA and RNA, so that the specificity of the DNA / RNA complex is improved, and the specificity of the DNA / RNA complex is improved. The kit can be better recognized and combined by an anti-DNA / RNA complex antibody, and has excellent detection specificity, sensitivity and precision.

This invention relates to the field of pathogen detection technology, specifically to an intelligent adaptive virus detection and optimization method and system. The method includes: acquiring raw sequence data generated by probe capture sequencing; performing real-time analysis of the sequence data using a deep learning model, outputting analysis results including the virus's presence state and sequence characteristics; dynamically adjusting control parameters of the hybridization capture process based on the analysis results through an adaptive control system; and completing hybridization capture according to the adjusted parameters and outputting a detection report. The system includes a sequencing data acquisition module, an analysis module, a sensor data acquisition module, an adaptive control module, and a detection report generation module. This invention, by combining artificial intelligence analysis with real-time process control, achieves intelligent optimization of the virus detection process, significantly improving the accuracy, sensitivity, and adaptability of detection. It is particularly suitable for detecting highly variable viruses and low-viral-load samples, and has significant clinical application value.

The invention belongs to the technical field of gene sequencing, and particularly relates to a library building kit and a probe for detecting hematologic tumor chemotherapy related gene polymorphism by using a hybrid capture method. According to the present invention, the genotype of the hematologic tumor chemotherapy related gene, the drug treatment effect, the toxicity and the metabolic capability related information can be rapidly obtained through the one-time detection; meanwhile, the library building kit is simple and convenient to operate and short in consumed time, and automatic detection is extremely easy to realize. The detection result completed by using the library building kit is accurate, and the library building kit has important reference significance for treatment and prognosis of hematologic tumor patients needing chemotherapy.

This invention provides a DNA hybridization capture device with a positioning component. The DNA hybridization capture device with a positioning component includes: a rectangular column; sliding components are installed on the outer walls of the left and right sides of the rectangular column; the sliding components are connected to a supporting component; the supporting component is connected to a clamping and positioning component; and the clamping and positioning component is connected to the capture device body. The DNA hybridization capture device with a positioning component provided by this invention allows the rotating shaft to rotate via a worm gear, which in turn drives a worm wheel to rotate. The worm wheel then moves a rectangular plate along a rack, allowing the capture device body to be moved to a suitable height to accommodate workers of different heights. The worm gear and worm wheel also provide a self-locking function when the handle is not rotated. When the capture device body is placed on the supporting plate, the output of the motor drives a bidirectional threaded rod to rotate, causing two sets of threaded blocks to simultaneously move the clamping plate towards the outer wall of the capture device body until the capture device body is stably clamped.

The invention relates to an ovarian cancer targeted medication related gene NGS detection kit and application thereof, and the kit comprises a probe group used for detecting ovarian cancer targeted medication related gene variation conditions and corresponding reagents based on a next generation sequencing technology. By means of a hybrid capture method, variation site conditions of 12 genes (BRAF, NTRK1, NTRK2, NTRK3, RET, MLH1, MSH2, MSH6, PMS2, BRCA1, BRCA2 and KRAS) related to important targeted medication of ovarian cancer can be detected at a time. The probe used in the invention has the advantages of wide coverage, high sequencing depth and strong specificity to ovarian cancer patients, and compared with polygene solid tumor large panel detection, the resource waste caused by irrelevant gene detection is reduced, and the reduction of the detection cost and the improvement of the detection efficiency are realized.

The invention discloses a nucleic acid detection method based on patterning, and belongs to the technical field of molecular diagnosis and biosensing. According to the method, a spatial position coded DNA framework is used as a detection carrier, specific capture probes are fixed at a plurality of pre-designed independent sites, a compound formed by target nucleic acid and a biotinylation report chain is subjected to specific hybridization capture, and then a recognizable signal is formed through combination of streptavidin; and the synchronous digital detection of multiple target nucleic acids is realized by reading patterned signals through an atomic force microscope. The position programmability and the single molecule detection sensitivity of the DNA nanotechnology are combined, the multiple detection capacity is only limited by the number of independent sites of a DNA frame, the expandability is extremely high, and the method is suitable for high-specificity detection of multiple nucleic acids in complex biological samples.

The present disclosure provides a method for enriching a nucleic acid molecule.

The invention discloses a site, a primer, a kit and a method for detecting cervical cancer DNA methylation, a DNA methylation site LRFN5 is obtained through library construction, hybrid capture, high-throughput sequencing and qMSP detection on a cervical cancer tissue sample through screening, and it is found that the LRFN5 site has differentiated expression in the early stage (1-2 stages) of CIN compared with cervical cancer samples in other stages, so that the DNA methylation site LRFN5 can be used for detecting cervical cancer DNA methylation. Therefore, the site can be used for early recognition of the cervical cancer, and early diagnosis and early treatment of the cervical cancer are facilitated.

The application relates to a target nucleic acid capturing method and a detection method, which comprises the following steps: concentrating a nucleic acid library to be detected to obtain a nucleic acid library to be detected concentrate; mixing and dissolving the nucleic acid library to be detected concentrate with a probe group freeze-dried product to obtain a hybridization system, and performing a hybridization reaction; and after the hybridization reaction, adding capturing magnetic beads to capture the probe group, and obtaining a target nucleic acid library; wherein the probe group contains a base sequence which is at least partially complementary to the target nucleic acid. The method can improve the performance of hybridization capturing.

The application discloses a probe and a method for detecting micro residual lesions. The probe for detecting micro residual lesions is a four-layer encrypted imbricated hybrid capture probe, wherein two layers of probes are wild-type probes, and two layers are mutant probes. The nucleic acid sequence of the wild-type probe comprises the sequence described in SEQ ID NO. 1-SEQ ID NO. 74, and the nucleic acid sequence of the mutant probe comprises the sequence described in SEQ ID NO. 75-SEQ ID NO. 149. The four-layer encrypted imbricated hybrid capture probe for detecting micro residual lesions is designed, a micro residual lesion molecular detection method with high sensitivity and good specificity is provided, the low-frequency mutation capture efficiency is high, and the detection limit of the ct DNA mutation frequency containing micro residual lesions can be as low as 0.005%.

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.

The application discloses a mutation background noise filtering calculation method based on second-generation sequencing data, and relates to the technical field of high-throughput sequencing data analysis, to solve the problems that the prior art depends on a baseline, has poor adaptability and high false positives. The algorithm first preprocesses sequencing data to obtain a pileup file, classifies sites according to sequence context and calculates error rates, constructs a hierarchical background noise model to test mutations, combines sample types to qualitatively determine real mutations and background noise, and simultaneously evaluates sample quality. The algorithm does not need to construct a baseline in advance, is suitable for hybrid capture and amplicon sequencing, can improve mutation detection specificity and consistency of repeated samples, guarantees reliable results, and is suitable for tumor gene detection and other scenes.

The invention discloses a probe group for targeted capture and detection of 65 gene variation related to oral and maxillofacial tooth / bone-derived lesions. The probe group comprises sequences as shown in SED ID No. 1 to SED ID No. 130. The invention also provides a kit containing the probe group, and application of the kit in preparation of products for targeted capture and detection of oral and maxillofacial tooth / bone-derived lesion related genes. The invention also provides a method for constructing an oral maxillofacial tooth / bone-derived lesion related gene detection model by using the probe group, and the method comprises the following steps: preparing a pre-library, preparing a probe hybridization capture library, sequencing, analyzing a sequencing result, and classifying obtained mutation sites to construct the model. According to the obtained sequencing result and model, the oral and maxillofacial tooth / bone-derived lesions which are difficult to distinguish in histomorphology can be accurately classified, accurate pathological diagnosis can be conveniently obtained, and help is provided for subsequent treatment and prognosis evaluation of patients.

According to the kit for detecting the inherited metabolic disease related genes based on the hybridization capture method, the hybridization capture method and the application, the kit comprises a probe pool of capture probes, and the capture probes are used for hybridizing and capturing the inherited metabolic disease related genes. The kit disclosed by the invention has the beneficial effects that when the kit is used for constructing a DNA library, the initial quantity of a template can be as low as 10ng, and enzyme digestion breaking, terminal filling and A tail adding are completed in one system, so that the consumed time is short, and the efficiency is high; secondly, the number of index pairs of the kit is up to 384, and 384 libraries can be constructed at the same time and can be used for sequencing on a machine together; in addition, according to a rapid hybridization system of the kit, the shortest hybridization time can reach 15 min, and the efficiency of common overnight hybridization can be achieved. The size of the finally obtained library fragment is about 300-500bp, and the library fragment can be compatible with various sequencing platforms of illumina for sequencing.

The preparation of the rheumatic immune disease drug therapy gene detection kit comprises four steps of nucleic acid extraction, library construction, hybridization and capture, in the nucleic acid extraction, a nucleic acid extraction and purification kit is adopted to carry out genome DNA extraction on a human EDTA anticoagulant peripheral blood sample; library construction comprises six sub-steps of enzyme digestion breaking, terminal repairing, 3'end A addition and joint connection, purification after connection, PCR amplification and purification after PCR amplification. The hybridization comprises two sub-steps of preparation before test and library and probe hybridization; the capture comprises five sub-steps of preparation before an experiment, target area DNA capture, PCR amplification after capture, purification after amplification, and capture library quality control. The detection method for preparing the rheumatic immune disease drug therapy gene detection kit comprises the following steps: firstly, carrying out on-machine sequencing, and then carrying out biological information analysis. The kit prepared by the invention can be used for detecting the variation of the gene loci including the probe based on a human peripheral blood sample, so that the diagnosis efficiency of medication of the rheumatic immune disease is effectively improved.

Provided are a micro-satellite instability detection method and apparatus based on a next-generation sequencing technology, belonging to the technical field of biological detection. The method comprises: selecting MSI sites, and designing capture probes and / or amplicon primers on the basis of any combination of the sites; selecting a plurality of samples, and calculating the proportion of deletion fragments and the proportion of normal fragments at each MSI site for each sample; selecting a plurality of samples that are determined to be in an MSI-L or MSS state by a PCR method as baseline samples, and setting different depth levels to construct baselines for each baseline sample; performing MSI state detection; selecting samples with an MSI state of MSI-H, and determining whether the samples are abnormal samples by using a Spearman rank correlation coefficient; and if yes, adjusting the baselines and re-determining the MSI state. The provided method has high stability and high accuracy, does not require a control sample, and can be simultaneously applied to micro-satellite instability detection for hybrid capture and amplicon sequencing data.

The invention discloses a pseudo-ginseng genome breeding chip based on a liquid phase hybridization capture technology and a development method thereof. The development method comprises the following steps: S1, confirming a target area and screening sites; s2, designing an intelligent probe; s3, efficient capture detection and verification; and S4, application service. The invention provides a liquid phase chip technical solution which is low in cost, high in flexibility, high in precision, high in usability and concentrated in function. According to the technology, through a self-developed probe design strategy, the capture efficiency of a complex genome region is remarkably improved, and the technical indexes that the coverage degree is not lower than 97% and the uniformity is not lower than 90% are achieved. And by combining a DNBSEQ-T7 high-throughput sequencing platform, the overall sequencing throughput and data quality are effectively improved. Meanwhile, the scheme supports site customization design, capture sites can be flexibly configured according to different varieties, characters or breeding targets, and good expansibility and adaptability are achieved.

The invention discloses a library construction method for detecting gastrointestinal stromal tumor molecular typing and a gastrointestinal stromal tumor molecular typing method, and belongs to the technical field of biology. Comprising the following steps: fragmenting a DNA (Deoxyribonucleic Acid) extract of a gastrointestinal stromal tumor patient specimen, modifying a terminal, connecting a joint, purifying and pre-amplifying, and carrying out hybridization reaction on a hybridization capture enrichment probe and a pre-amplified sequence to enrich a target sequence; the hybridization capture enrichment probe comprises a sequence as shown in SEQ ID NO: 1-109; the method comprises the following steps of: performing amplification after non-specific capture and binding DNA fragments are eluted and removed, performing nucleic acid concentration and fragment length quantification on a product to obtain a library, and performing bioinformatics analysis on gastrointestinal stromal tumor patient specimen data by next-generation sequencing to obtain gastrointestinal stromal tumor molecular typing information. According to the invention, in-vitro non-diagnostic gastrointestinal stromal tumor molecular typing joint detection is realized, multi-gene variation form identification is completed in one experiment, the detection cost is saved, and the sensitivity is high.