344 results about "Fluorescence in situ hybridization" patented technology

The present invention relates to a novel method for the amplification of DNA, this method being particularly useful for the amplification of the DNA or the whole genome of a single cell, chromosomes or fragments thereof. Described is also the use of the method in DNA analysis for medical, forensic, diagnostic or scientific purposes, like comparative genomic hybridization (CGH)-, fluorescence in situ hybridization (FISH)-, polymerase chain reaction (PCR)-, single strand conformation polymorphism (SSCP)-, DNA sequence-, "loss of heterozygosity" (LOH)-, fingerprint- and/or restriction fragment length polymorphism (RFLP)-analysis.

This invention relates to methods of analyzing a tissue sample from a subject. In particular, the invention combines morphological staining and/or immunohistochemistry (IHC) with fluorescence in situ hybridization (FISH) within the same section of a tissue sample. The analysis can be automated or manual. The invention also relates to kits for use in the above methods.

Amplification and overexpression of theHER-2 oncogene in breast cancer is felt to be stable over the course of disease and concordant between the primary tumor and metastases. Therefore, patients with HER-2 negative primary tumors will rarely receive anti-HER-2 antibody therapy. A very sensitive blood test is used to capture circulating tumor cells (CTC's) and evaluate their HER-2 gene status by FISH evaluation. The HER-2 status of the primary tumor and corresponding CTC's is used to assess the ratio of CTC's as a reliable surrogate marker. HER-2 expression of 10 CTC's is sufficient to make a definitive diagnosis of the HER-2 gene status for the whole population of CTC's in patients with recurrent breast cancer.

The present invention provides methods for preparing cells with highly condensed chromosomes, such as sperm, and methods for detecting and quantifying specific cellular target molecules in intact cells. Specifically, methods are provided for detecting chromosomes and chromosomal abnormalities, including aneuploidy, in intact cells using fluorescence in situ hybridization of cells in suspension, such as sperm cells.

The invention provides a fluorescence in situ hybridization (FISH) image parallel processing and analysis method. The method employs parallel processing, detection of chromosome fluorescent labeling points and edge detection and segmentation of nucleuses can be performed simultaneously, and the processing time is reduced; and a watershed algorithm based on adaptive shape labels is employed so that the segmentation precision of the adhered nucleuses in FISH images is substantially improved, the detection accuracy of relative positions of the nucleuses in tumor cells and the chromosome fluorescent label points is improved, and online real-time detection of the tumor cells can be realized.

The invention relates to a preparation method for probes related to breast cancer molecular markers and to preparation of a breast cancer fluorescence in situ hybridization detection kit by using the probes. The breast cancer fluorescence in situ hybridization detection kit can be prepared from HER2, TOP2A and AGTR1 gene probes prepared by using the method provided in the invention, human chromosome 17 counting probe, hybridization buffer, unlabelled competitive DNA and DAPI counter strain; application of the kit enables the detection rate of breast cancers to be substantially improved and type sorting of breast cancers to be more accurate and provides guidance to formulation of an individual therapeutic schedule and selection of proper therapeutic drugs, thereby lowering down mortality, reducing recurrence risk and achieving the goal of optimizing the effect of diagnosis and treatment.

An embodiment is disclosed for performing the image processing for analyzing the results of a fluorescence in situ hybridization (FISH) microscopic automated sample analysis to determine specific chromosomal characteristics.

The invention belongs to the field of biotechnology, and discloses a FISH (fluorescence in situ hybridization) probe, a kit and a detection method for detecting Her2 (human epidermal growth factor receptor 2) gene free from repetitive sequence. The Her2 gene is used as a template to perform polymerase chain reaction on non-repetitive sequence in the Her2 gene, the amplification product is DNA (deoxyribonucleic acid) fragments with 350-800 different base pairs, the repetitive sequence in the Her2 gene is eliminated so as to form the product free from repetitive sequence; after the product is in fluorescence labeling, the Her2 gene free from repetitive FISH is obtained for the Her2 gene FISH detection. The Her2 gene FISH probe obtained by the invention can eliminate the repetitive sequence, the non-specific background signal of the Her2 gene FISH probe can be obviously reduced, and the specificity of the FISH probe is improved.

The invention provides a breeding method for the germ plasm of a new translocation line wheat-rye T2BL.1RS, which is characterized in that, distant hybridization is performed by taking common wheat variety Xiaoyan 6 as female parent and relative plant rye variety German White as male parent; seeds are acquired after immature embryo salvation culture, colchicine half root soaking method and chromosome doubling; backcross seeds are acquired after disease resistance identification, cytological identification and backcrossing utilizing the original parent Xiaoyan 6 as male parent; then after disease resistance identification and cytological identification, individual plants with excellent comprehensive characters are selected in direction for self crossing, and after continuous self cross selective breeding for six generations, the finally acquired individual plant is confirmed to be wheat-rye T2BL.1RS translocation line after identified by continuous total genome in situ hybridization and three-probe multicolor fluorescence in situ hybridization. The breeding method for the germ plasm of a new translocation line wheat-rye T2BL.1RS has the advantages that, the wheat-rye T2BL.1RS translocation line has high resistance to stripe rust, current prevalent fungus strain, powdery mildew and other important virus strains; the invention has excellent high yield character; the method lays an excellent germplasm base for the cultivation of breakthrough new wheat variety with disease resistance and high yield.

The invention provides a method for identifying exogenous chromosomes and chromosome segments in plant distant hybrids. The method includes the steps of firstly, microscopically separating monosomes or chromosome segments of chromosomes to be tested of a plant to be tested; secondly, using the separated monosomes or chromosome segments in the first step to build a fluorescence probe of the monosomes or chromosome segments; thirdly, using the fluorescence probe built in the second step to perform fluorescence in situ hybridization to chromosomes in metaphase division of the hybrids; and fourthly, analyzing the fluorescence in situ hybridization result obtained in the third step to determine distribution of fluorescence signals of the chromosomes to be tested of the plant to be tested. Exogenous chromosomes and chromosome segments in wheat distant hybrids can be identified by the method, and the method is applicable to identification of exogenous chromosomes and chromosome segments in distant hybrids of other plants.

The invention discloses a method for obtaining an oligonucleotide probe. The method for obtaining the oligonucleotide probe comprises the following steps: downloading original sequences of pSc119.2, pAs1, pTa-535, pTa71, pAWRC.1 and CCS1 from a national center of biotechnology information (NCBI) website; analyzing the downloaded sequences by using bioinformatics software DNAMAN; seeking repeated core units in these sequences; returning to the NCBI website to compare by using a blastn tool for finding out a core oligonucleotide sequence of which the length is below 60bp to carry out probe synthesis; carrying out probe synthesis on a lot of oligonucleotide sequences which are primarily screened out; gradually carrying out a fluorescence in situ hybridization (FISH) test after probe synthesis; verifying the function of the probe, and verifying whether the synthetic oligonucleotide sequence can play the FISH effects of pSc119.2, pAs1, pTa-535, pTa71, pAWRC.1 and CCS1 or not, so as to obtain the oligonucleotide probe. By adopting the method disclosed by the invention, complicated procedures of the existing probe marking technique are removed, the problem of marking failure of the probe is removed, and the cost is reduced.

The present invention comprises a method of detecting circular tumor cells and methods of detecting, evaluating, or staging cancer in a patient, as well as a method of monitoring treatment of cancer in a patient using the claimed method. The method comprises contacting a sample with a CD45 binding agent; selecting the cells based on positive or negative CD45 staining; contacting the selected cells with a labeled nucleic acid probe, and detecting hybridized cells by fluorescence in situ hybridization; and analyzing a signal produced by the labels on the hybridized cells to detect the CTCs. In other embodiments, the method provides for directed to a method of determining the level of CTCs in a sample having blood cells from a patient by contacting a sample having blood cells from a patient, wherein the sample has not been pre-sorted into CD45-positive and CD45-negative cells.

The invention belongs to the field of biotechnology, and discloses a FISH (fluorescence in situ hybridization) probe, a kit and a detection method for detecting BCR/ABL fusion gene free from repetitive sequence. A BCR gene and ABL gene are used as templates to perform polymerase chain reaction on non-repetitive sequence in the BCR gene and ABL gene, the amplification product is DNA (deoxyribonucleic acid) fragments with 350-800 different base-pairs, the repetitive sequences in the BCR gene and ABL gene are eliminated so as to form the product free from repetitive sequence; after the product is in fluorescence labeling, the BCR gene and ABL gene free from repetitive FISH are obtained for the BCR gene and ABL gene FISH detection. The BCR/ABL fusion gene FISH probe obtained by the invention can be used for eliminating the repetitive sequence, the non-specific background signal of the FISH probe can be obviously reduced, and the specificity of the FISH probe is improved.

The invention belongs to the application field of fluorescence in situ hybridization probes and discloses a probe set for diagnosing Xp11.2 ectopic perivascular epithelioid cell neoplasm and application of the probe set. The probe set includes BAC clonal fragments RP11-918B12, RP11-916D13, RP11-416B14 and RP11-344N17. The probe set is capable of diagnosing the specificity and the sensibility of Xp11.2 ectopic PEComa 100%, is convenient, quick and reliable, and can ensure a high success rate. And moreover, the probe set can be used for preparing an Xp11.2 ectopic PEComa diagnostic kit.

The invention discloses a method for detecting polymorphism of a flora of a prawn culture water body. The method comprises the following steps: (1) extracting total genomic DNA of mixed microbes in a prawn culture water body sample; (2) designing a specificity T-RFLP-PCR universal primer and performing PCR circulating reaction; (3) purifying a product and performing enzyme cutting on DNA by a specific restriction enzyme Hae III; (4) performing ionophortic separation on DNA segments by 1 percent agarose gel, performing fluorescent scanning on the DNA segments; (5) analyzing a polymorphism structure of a microbe flora; and (6) performing quantitative detection on predominant bacteria of the flora through fluorescence in situ hybridization technology. The method improves detection technology combining T-RFLP-PCR with FISH, uses fluorescence to mark a specific primer; and compared with the modern technology, the method has the characteristics of high repeatability, sensitivity, rapidness, accuracy, stability and the like, and can qualitatively and quantitatively analyze the ecological diversity of the microbes in the prawn culture water body along with time change and the dynamic change of a composite structure of the predominant flora.

Gene probes for specific regions of chromosomes 1, 3, 9, 10 and 17 are now shown to be useful in the diagnosis and prognosis of smoking related cancers such as non-small cell lung cancer (NSCLC). For example, these probes can be used with fluorescence in situ hybridization (FISH), and used to stratify smokers into high and low risk groups, to determine susceptibility to the development of smoking related cancers, to predict cancer progression and treatment efficacy, and to rule out other diseases.

The method relates to a method for the highly fine three-dimensional modeling of complex earth surface, belonging to the technical field of analogue computation of rock-soil values. The technical scheme is that the method comprises the following steps of (1) obtaining an earth surface contour map and elevation data and establishing a three-dimensional earth surface wireframe; (2) importing the earth surface contour map and elevation data and the three-dimensional earth surface wireframe into 3DMine (Three-dimensional Mine) mineral engineering design software to build an earth surface solid model; (3) dividing earth surface grid points according to actual engineering demands to obtain grid point coordinate data; (4) exporting the grid point coordinate data and grouping and listing the grid point coordinate data in FLAC3D (Fast Lagrangian Analysis of Continua 3D); and (5) formulating a circulation program by utilizing a programming language FISH (Fluorescence In Situ Hybridization) embedded in the FLAC3D, reading a grid point coordinate data list and build an earth surface model. The method for the highly fine three-dimensional modeling of the complex earth surface has the advantages and effects that the self modeling function of the FLAC3D software is fully utilized, the simple practical modeling program is formulated through the utilization of the FISH language, the modeling procedure is simplified greatly, the modeling efficiency is improved, the modeling precision can be flexibly adjusted according to the actual engineering situation and the modeling fineness is high.

The invention provides a multicolor fluorescence in situ hybridization (MFISH) method for quickly analyzing and identifying alien chromosome of wheat. The method comprises the following steps: (1) preprocessing actively dividing tissues in meristematic zone of wheat root tip by N2O and conducting enzymolysis on the actively dividing tissues to obtain a metaphase specimen with clear images and uniform distribution of chromosomes; (2) marking a fluorescent probe of fundamental genome of wheat by nick translation method; (3) conducting fluorescence in situ hybridization on chromosomes at mitosis metaphase obtained in step (1) by using the constructed fluorescent probe in step (2); and (4) analyzing the fluorescence in situ hybridization results obtained in step (3). According to the method provided by the invention, not only can alien chromosomes and chromosome segments in distant hybrid wheat be identified, but also alien chromosomes and chromosome segments of distant hybrids of other crops can be identified.

The invention discloses a fluorescence in situ hybridization probe and kit for detecting HER-2 (human epidermal growth factor receptor-2) gene amplification and relates to an FISH (fluorescence in situ hybridization) kit. The fluorescence in situ hybridization kit for detecting HER-2 gene amplification comprises an LSP HER-2/CSP 17 probe and DAPI redyeing liquid; and the probe is marked by adopting a random primer method and is a fluorescence in situ hybridization double-color probe, an HER-2 gene is indicated by a red signal, and the internal control CSP17 is indicated by a green signal. An FFPE tissue slice is used as a detection sample, the FISH method is adopted for detecting the HER-2 gene amplification state, and the probe and the kit can be used for diagnosis of molecular markers for invasive breast cancer and stomach cancer and assisting in clinical medication selection.

The present invention relates to a gene probe composition and a kit for acute lymphocytic leukemia detection. The gene probe composition comprises a TEL gene probe, an AML1 gene probe, a PAX5 gene probe, a P16 gene probe and an IKZF1 gene probe. The present invention further provides an acute lymphocytic leukemia fluorescence in situ hybridization detection kit, wherein the kit comprises the gene probe composition. With the kit, concurrent five gene detection can be performed on the same sample even the single leukemia cell, and detection capability and efficiency are significantly improved.

The invention discloses a fluorescence in situ hybridization method for positioning the 45S rDNA on a plant chromosome. The fluorescence in situ hybridization method mainly comprises the following steps of preparation of a chromosome specimen, preparation of 45S rDNA probes, fluorescence in situ hybridization of the chromosome and hybridization signal detection. The provided method for probe marking and hybridization is simple, convenient, quick and accurate, and mainly solves the problem about complex and cumbersome marking of the existing probes for fluorescence in situ hybridization. The probes provided by the invention are oligonucleotide probes which are not marked any more, and a fluorophore for modification can be added during synthesis of the probes, and compared with the prior art, the method is simple and the cost is low. By utilizing the technology, FISH positioning of the 45S rDNA on the chromosome can be completed within 3 hours.

The invention provides methods for the rapid determination of the antibiotic susceptibility of a microorganism, such as, an infectious microorganism in a biological sample, using fluorescence in situ hybridization (“FISH”). Methods of the invention may be applied to the rapid identification, typing, antibiotic susceptibility determination, and/or antibiotic minimum inhibitory concentration (MIC) determination for any infectious microorganism, such as a Gram positive bacteria, a Gram negative bacteria, or a yeast.