50 results about "Oligonucleotide chip" patented technology

A method for preparing oligonucleotide chip for transgene product detection and use are disclosed. It includes: designing specific oligonucleotide probe and related primer by heterogeneric inserting gene, species internal standard gene, specific boundary sequence in transgene product gene set, fixing probe on glass slide to form transgene product detecting chip, amplifying DNA of plant to be tested by related primmer, marking by fluorescence, and hybridizing with chip. It can be use to acquire variable information.

A high-throughput method for rapid detection of virus pathogens comprises the following steps of: (1) designing probes and spotting on a chip to obtain an oligonucleotide chip; (2) designing and synthesizing primers to obtain primers corresponding to a detected pathogen; (3) performing synchronous asymmetric PCR amplification to a detected sample with the primers corresponding to the detected pathogen to obtain a single-stranded DNA product specifically complementary with the oligonucleotide; (4) labeling a fluorescent substance on the single-stranded DNA product to obtain a fluorescent-labeled single-stranded DNA product; (5) hybridizing with the oligonucleotide chip, and cleaning the chip; and (6) scanning with a fluorescent scanner to obtain virus detection and analysis results. The invention solves the time-consuming, labor-consuming, low sensibility and specificity and inaccurate detection result problems in the detection method of virus causing porcine epidemic diseases in the prior art. The invention can be used for rapid, accurate and efficient detection of multiple genes.

The invention relates to preparation and application of a gene chip capable of detecting adenoviruses in a parting mode.A preparation method comprises the steps of preparing specific primers and probes for different types of adenoviruses, preparing oligonucleotides chips, establishing a plurality of PCR systems, and establishing a hybridization system.The gene chip prepared through the method can screen different types of adenoviruses including a 3-type adenovirus, a 7-type adenovirus, a 14-type adenovirus, an 11-type adenovirus and a 55-type adenovirus.The gene chip has the advantages of quickness, accuracy, high throughput and high specificity.A new detection means is provided for clinical diagnoses of different types of adenovirus infections and epidemiological surveys.

The invention relates to preparation and application of a rickettsia discrimination detection gene chip. The preparation method comprises the following steps: preparing universal and specific primers, preparing seven rickettsia nucleic acid typing probes, preparing an oligonucleotide chip, establishing a multiplex PCR (polymerase chain reaction) system, and establishing a hybridization system. The gene chip can simultaneously discriminate seven important rickettsiae, including rickettsia prowazeki, rickettsia mooseri, spotted fever group rickettsia, Coxiella burnetii, Orientia tsutsugamushi, Ehrlichia chaffeensis and Anaplasma phagocytophilum. The gene chip has the advantages of high speed, high accuracy, high flux and high specificity, and can provide a novel detection means for clinical diagnosis and epidemiological investigation of rickettsia infection.

The invention discloses an oligonucleotide chip capable of detecting five enteroviruses simultaneously and an application thereof. The chip comprises a substrate, probes of pathogens of five enteroviruses coated on the substrate, negative contrast, positive contrast and blank contrast, wherein the probes of pathogens are five 54-70mer of HAV-P, ROV-P, NOV-P, ASV-P and ADV-PV1 probes. The chip of the invention adopts multiple PCR to amplify a plurality of target sequences of pathogens simultaneously and uses the downstream primer Tamara labeling method to perform fluorescence labeling to PCR product, the labeled PCR product is hybridized with the chip to realize the accurate detection of pathogens, the detection sensitivity is equal to that of PCR and the specificity is high. The detectionefficiency and accuracy are obviously shortened. The technology system of the invention is applicable to fields such as sea water and marine life specimens monitoring, food hygiene surveillance, customs quarantine, related clinical detection and the like.

The invention discloses an oligonucleotide chip labeling a random primer PCR amplification based on a biotin-avidin-cy3 reaction system, which is made by a substrate, and CMV, HSVI, RV, EBV or TB pathogen probes, a negative control, a positive control and a blank control that are coated on the substrate in an array way, wherein the substrate is a surface aldehyde glass slide; the pathogen probes are 10 of CMV-1, CMV-2, HSVI-1, HSVI-2, RV1, RV2, EBV1, EBV2, TB1 and TB2 probes with 70mer; the positive control is a synthesized marker probe; and the blank control is double distilled water. The invention performs the biotin-11-dUTP amplified labeling to various pathogen target sequences by using the random primer and detects the pathogen by adopting a method of increasing the labeling efficiency through using a biotin-avidin-cy3 signal amplification system, has high detection sensitivity and obviously shortens the detection cost and the detection time. The technical system can be suitable to the fields of clinical detection, health supervision, customs quarantine control, scientific research, and the like.

The invention relates to a gene chip for detecting important respiratory pathogenic viruses. A preparation method of the gene chip comprises the following steps of: preparing a specific primer; preparing a virus specific probe; preparing an oligonucleotide chip; establishing an RT-PCR (Reverse Transcription-Polymerase Chain Reaction) system; establishing a hybrid system; preparing a visual detection reagent; and establishing a developing method. The gene chip prepared by the invention can be used for simultaneously discriminating nine general respiratory pathogenic viruses comprising A and B type influenza viruses, parainfluenza viruses type 1 and 2, human metapneumovirus, respiratory syncytial virus, measles virus, rubella virus and mumps virus, provides a new solution for quickly detecting the general respiratory pathogenic viruses at high throughout and can provide guidance for monitoring, clinically diagnosing and treating the respiratory pathogenic viruses.

The invention discloses a space transcriptome sequencing technology device based on tissue and chip space position information. The device comprises a cover slip and a silicon-based oligonucleotide chip, wherein an anchoring probe is arranged on the silicon-based oligonucleotide chip; a gap between the cover slip and the silicon-based oligonucleotide chip is used for accommodating a tissue freezing section with the thickness of 10-50 um. Meanwhile, the invention further provides a space transcriptome sequencing technology method adopting the device. According to the device and the method, sufficient RNA (ribonucleic acid) samples can be effectively acquired from a small quantity of cells, uniform or overall characterization of a sequencing result is avoided, further, difference of different phenotypes of cell transcript information in tissue can be distinguished according to cell heterogeneity, and transcript information with low abundance is mined, so that that more accurate transcript information with high specificity is provided for fields of biology, medicine and agriculture.

The invention relates to a sol-gel process for the functionalisation of a surface of a solid substrate in order to provide said surface with chemical functions that can react with chemical or biological molecules, i.e. chemical functions that can immobilise, attach, graft biological or chemical molecules. The invention can be used, for example, in the preparation of chemical and biological sensors, such as for the production of chips comprising DNA, oligonucleotides, sugars, peptides and small organic molecules.

The invention relates to a diagnostic kit for identification of mycobacterium strains and detection of drug resistance and a preparation method thereof. The kit can quickly and accurately distinguish between the identification of mycobacterium strains and the detection of drug resistance in large quantities by using an oligonucleotide chip. Point mutations in the rpoB gene of Mycobacterium tuberculosis associated with drug resistance. The diagnostic kit for the identification of mycobacterium strains and the detection of drug resistance of the present invention contains an oligonucleotide chip, and the chip contains a complex composed of mycobacterium tuberculosis mycobacterium strain identification probes and drug resistance detection probes. Probe, mycobacterium tuberculosis rpoB gene, and fluorescent material, wherein the fluorescent material contains biotin-binding protein, which is used as the label of biotin and the corresponding probe to detect the hybridization of the sample amplification product.

A process for preparing oligonucleotide chip used in DNA surface calculation to solve the NP question is disclosed, which features that the coding and arranging of the oligonucleotide sequenes on DNA surface calculation chip are in such manner that the data in data pool is converted to the binary form, the data pool is converted to an array, and each data in the array is mapped to an oligonucleotide sequence, that is, a point on the chip.

The invention relates to a hemorrhagic fever associated pathogen identifying gene chip; the preparation method comprises preparation of a specific primer, preparation of a pathogen specific oligonucleotide probe, preparation of an oligonucleotide chip, establishment of an RT-PCR (reverse transcription-polymerase chain reaction) system and establishment of a hybrid system and a signal detection method. The gene chip prepared by the invention can be used for simultaneously identifying 16 hemorrhagic fever associated pathogen microorganisms, including Zaire Ebola virus, Sudan Ebola virus, marburg virus, lassa virus, junin virus, Machupo virus, rift valley fever virus, Crimea-Congo hemorrhagic fever virus, plasmodium, hantaan virus, SFTS (severe fever with thrombocytopenia syndrome) virus, dengue virus, yellow fever virus, Chikungunya virus, influenza A virus and influenza B virus. The gene chip has the characteristics of being rapid and accurate, high in throughput and high in sensitivity; and a new technological means is offered for the diagnosis of hemorrhagic fever pathogen, health supervision and the control and prevention of infectious diseases.

The invention discloses a detection method synthesizing microfluidic chips in situ, which comprises the following steps that: 1) a sequence of non-coding small RNAs reported on the prior model plant is utilized, sequence information of miRNA is preferably selected, and complementary oligonucleotide hybridization probes are designed according to the sequence of a sense strand or an antisense strand of miRNA; 2) an oligonucleotide chip is prepared by an in-site synthetic method; 3) small RNAs smaller than a 50 oligonucleotide group are extracted from a plant to be detected, then are hybridized with the probes in the chip after making end labeling, and the result of chip hybridization is obtained to perform qualitative and quantitative analysis. The detection method can be used for the validation of miRNA (and other sNC-RNAs) obtained through the forecast by a bioinformatics method, can also be used for the detection and identification of miRNA (and other sNC-RNAs) in new plant species, and can also be used for the determination of miRNA (and other sNC-RNAs) variation relation under different treatment conditions (eg. under the condition of virus infection).

An oligonucleotide chip for detecting the mutation sites of hepatitis B virus is composed of glass substrate, oligonucleotide probes array and reference point-type coated laeyr. The said probes arrayhas 72 probes, including 12 known mutation sites and relative wild sites. The process for detcting mutation sites of hepatitis B virus includes such steps as mixing the probes as solid primer, hepatitis B virus DNA and template, and three pairs of liquid primers, PCR amplification via fluorescence labeled Cy3-dCTP, and analyzing mutation sites according to intensity of fluorescent signal. Its advantages are high speed, efficiency and correctness.

The invention relates to a preparation method and application of a VRE/MRSA/KPC/NDM-1 detection gene chip. The preparation method includes: preparing specific primers, preparing specific probes, preparing an oligonucleotide chip, and building a multiple PCR system and a hybrid system. By the prepared gene chip, 5 drug-resistant genes and 3 related pathogenic bacteria can be detected at the same time, and the 5 drug-resistant genes and 3 related pathogenic bacteria include Vancomycin-resistant Enterococcus VRE drug-resistant genes VanA and VanB, an encoding New Delhi metallo-beta-lactamase-1 NDM-1 drug-resistant gene, a methicillin-resistant staphylococcus aureus MRSA drug-resistant gene mecA, an encoding carbapenemase KPC gene, an enterococcus faecalis specific gene Fddl, an enterococcus faecium specific gene Sddl and a staphylococcus aureus specific gene nuc. The chip which provides a new detection manner for the confirmation of the drug-resistant genes of common antibiotics resistance phenotypes has advantages of quickness, accuracy, high flux and the like.

The present invention relates to a β-catenin oligonucleotide microchip for detecting mutation in the mutational hot spot regions of β-catenin gene, a manufacturing process thereof and a method for detecting the β-catenin mutation employing same, wherein specific oligonucleotides are selectively designed to detect various missense mutations and in-frame deletion at the mutational hot spots of β-catenin gene. The β-catenin oligo chip of the present invention can be used in studies to detect β-catenin mutations and unravel the signal transduction mechanism and tumorigenesis related to β-catenin gene.

The invention relates to a guiding gene chip for HCV infection individual treatment. The preparation method of the guiding gene chip comprises the following steps: preparing a universal primer; preparing an HCV hypotype nucleic acid parting probe and a human IL-28B rs12979860 investigation of polymorphism probe; preparing an oligonucleotide chip; building a polyfunctional RT-PCR system; building a hybrid system. The gene chip prepared by the method can discriminate five hypotypes of 1b, 2a, 3a, 3b and 6a of HCV at the same time, can detect polymorphism of CC, TT and CT three genes of human rs12979860, has the advantages that the gene chip is quick, accurate, and high in flux and specificity, and can provide guiding for HCV clinical diagnosis and individual treatment.

The invention relates to a gene chip, and discloses an oligonucleotide chip for detecting a whole genome CpG island. The invention consists of solid-phase carriers and probes, wherein, the probes are hybridized with nucleotide sequences and/or complementary sequences thereof of the CpG island in the whole genome and 2kb areas of the upstream and downstream of the CpG island. The invention also discloses a method of detecting the position of the whole genome CpG island with the oligonucleotide chip. The oligonucleotide chip of the invention can carry out the CpG island detection of methylate and/or methylate DNA rapidly and accurately with high flux, and provides a powerful tool for correlation studies on the whole genome CpG island.

The invention belongs to the technical field of biology, and relates to a detection method for nucleic acid, in particular to a method for designing an oligonucleotide probe. The method for designing the oligonucleotide probe comprises the following steps of: (1) designing a target gene oligonucleotide probe with length of 50mer aiming at target genes; (2) designing general sequences, the homologies of which are not more than 40 percent compared with the target genes and the lengths of which are 10mer; and (3) connecting the general sequences at two ends of the target gene oligonucleotide probe to construct the oligonucleotide probe respectively. The sequence of the human immunodeficiency virus oligonucleotide probe constructed by adopting the design method of the invention is SEQ ID No: 1. Fluorescence labeled complementary sequences are obtained by fluorescence labeling of the complementary sequences of the obtained general sequences. The fluorescence labeled complementary sequences can be used for detecting the quality of an oligonucleotide chip, have the advantage of high sensitivity, and are particularly suitable for detection chip design of clinical pathogen genes.

High-throughput detection for the interesting base or the mutation site in the nucleic acid sample can be achieved by the linear test probe pairs P1 and P2. The test probe pairs P1 and P2 respectively comprise either of the flanking complementary sequences which are adjacent to the interesting base or the mutation site in the nucleic acid sample. When the test probe pairs P1, P2 are annealed and hybridized to the nucleic acid sample, a gap will be generated at the interesting base or the mutation site position between the probe pairs and the sample. Divide the annealed hybrid sample into four equal reaction systems to which add dATP, dTTP, dCTP, dGTP, respectively. The test probe pairs P1 and P2 will be ligated into one single probe when adding the complementary nucleotide system under the DNA polymerase or ligase. After purified and amplified, the generated single probes are hybridized to the corresponding area in a common oligonucleotide microarray. The generated single probe will give a signal in the hybrid area, and therefore detect and analyze the hybrid signal to determine the base type or the mutation genotype at the detection position. The invention can be applied to the re-sequencing the target nucleic acid sequence, the detection and analysis for the mutation, insertion, or deletion sites of a known nucleic acid sequence, and the genotyping of the pathogenic microorganism.

The invention provides an oligonucleotide chip for synchronous detection of four porcine diarrhea viruses. The oligonucleotide chip comprises a solid phase carrier and oligonucleotide probes fixed onthe solid phase carrier, wherein the oligonucleotide probes comprise a probe shown in SEQ ID No. 1-2, a probe shown in SEQ ID No. 3-4, a probe shown in SEQ ID No. 5-6, and a probe shown in SEQ ID No.7-8; the probes are respectively used for detecting porcine epidemic diarrhea virus (PEDV), porcine transmissible gastroenteritis virus (TGEV), porcine group A rotavirus (GAR) and porcine delta coronavirus (PDCoV). The invention also provides a kit and a method for the synchronous detection of the four porcine diarrhea viruses. The chip and the method can realize the high flux co-detection of thefour porcine diarrhea viruses, i.e., PEDV, TGEV, GAR and PDCoV, and can be applied to mass detection of clinical diseases, thus having a wide application prospect.

The invention discloses a method for detecting a transgenic tomato by using an in-situ synthetic microfluidic chip. The method comprises the following steps of: 1) designing a chip probe for detecting an exogenous gene of the transgenic tomato; 2) preparing an oligonucleotide chip; 3) extracting genome deoxyribonucleic acid (DNA) in a tomato sample to be detected; 4) marking a specific segment of the genome or the entire genome by a fluorescent agent; 5) chip hybridization; 6) judging whether the sample belongs to the transgenic tomato variety. By adopting the method, an oligonucleotide probe is designed according to the sequence information of the known tomato exogenous gene which has been publically reported; the microfluidic gene chip is prepared by utilizing an in-situ synthesis method, so as to achieve simultaneous detection of existence of the exogenous genes related to insect resistance, antiviral action, herbicide resistance, delayed fruit, softening resistance, storage resistance and the like on the same chip. Thus, an effective method is provided for rapid detection and identification of the transgenic ingredients of the tomato and highly processed products; a foundation is provided for building and perfecting a transgenosis detection system and a supervisory system.

The invention discloses an information storage method based on oligonucleotide chip hybridization, which comprises the following target information writing steps: (1) converting target information into a binary code, then segmenting the binary code into a plurality of fields, each field corresponding to a site on an oligonucleotide chip, and each site comprising N binary digits; (2) mapping the 1/0 value on the N binary digits of each site into a presence/absence combination of N oligonucleotide coding chains to obtain a combination list of oligonucleotide coding chains needing to be added to all the sites on the oligonucleotide chip; and (3) fixing the oligonucleotide coding chains in the sites on the surface of the oligonucleotide chip in a covalent bond manner according to the oligonucleotide coding chain combination list, thereby completing the writing of the target information on the oligonucleotide chip. According to the invention, the access of various complex file formats is realized, the parallelism is good, the signal-to-noise ratio is high, the access speed is high, and the accuracy is good.