493 results about "Molecular beacon" patented technology

A method for detecting compounds of interest in bodily fluids, including exhaled breath and blood. The present invention uses biosensors that mimic naturally occurring cellular mechanisms, including RNA oligonucleotide chains or “aptamers,” in combination with molecular beacons or nanotechnology to provide an effective and efficient method for diagnosing a condition and / or disease within a patient. The subject invention also provides a method for screening those analytes / biomarkers likely to be present in exhaled breath.

Provided are compositions that include an at least partially single-stranded nucleic acid, at least one first molecular beacon, and an enzyme comprising a helicase activity, which enzyme is capable of removing the first molecular beacon from the single-stranded nucleic acid wherein the first molecular beacon is hybridized to a first complementary subsequence of the nucleic acid. Also provided are methods of determining the sequence of a template nucleic acid that include removing molecular beacons that are hybridized to the template from the template in a sequential manner using an enzyme that exhibits a helicase activity, detecting a sequence of fluorescent signals that is produced by the removal of a molecular beacons, and converting the sequence of fluorescent signals into nucleotide sequence information. Sequencing systems in which compositions and methods of the invention can be used are also provided.

The invention belongs to the technical field of neoplastic disease diagnosis, and discloses a kit for early diagnosis of neoplastic disease, a method and applications of the kit. The kit includes a specific capturing and detecting system for a tumor exosome and a molecular beacon fluorescent detection system. The specific capturing and detecting system for the tumor exosome includes a solution ofa magnetic gold nanosphere composite modified by a tumor exosome specific ligand and adopting ferric ferrous oxide as a core, and a gold nanocage solution modified by the exosome specific ligand. Themolecular beacon fluorescent detection system includes a molecular beacon probe provided with a fluorescent group, fluorescent amplification substrate hybrid chains provided with fluorescent groups and a buffer liquid. The kit and the method combine tumor exosome specific capturing and analysis and a molecular beacon fluorescent detection method, and greatly increases accuracy and reliability of early diagnosis of tumor.

The invention discloses a cascading isothermal amplification based miRNA fluorescence detection kit and a miRNA fluorescence detection method. The target miRNA can specifically recognize and form a stable three-way crossing structure with a ternary probe through complementary hybridization of nucleic acids; a ternary primer in the three-way crossing structure performs strand displacement amplification (SDA) along a ternary template and generates a number of SDA template products; the SDA template product can open a difunctional hairpin probe to trigger nickase to perform signal amplification reaction finally; and a cyclophorase digestion molecular beacon can generate cascading amplified fluorescence signals. The kit can detect miRNA low to 100fM; and the detection method has superhigh specificity, can well distinguish highly homologous sequences at different mispairing positions and solves the difficult problem of false positive result caused by poor specificity in the existing detection method.

The invention discloses a multi-target quantum-dot mark nucleic acid chip and a preparation method and a detection method thereof, wherein the nucleic acid chip comprises a solid phase holder and an oligonucleotide probe array fixed on the surface of the solid phase holder, wherein the oligonucleotide probe array comprises at least two oligonucleotide probes which do not contain self-complementary sequences; one end of the oligonucleotide probe marks quantum dots and is fixed by the quantum dots, and the oligonucleotide probes with different sequences are marked by the quantum dots which emit fluorescence with different wavelengths; or the oligonucleotide probe array comprises at least two molecular beacons, wherein one end of the molecular beacon marks quantum dots and is fixed by the quantum dots, the molecular beacons with different sequences are marked by the quantum dots which emit fluorescent with different wavelengths, and the other end of the molecular beacon is marked by a fluorescence quenching group. By utilizing the basic-group complementation pairing principle and the FRET phenomenon, the detection of a plurality of special nucleic acid sequences in a nucleic acid sample to be detected can be simultaneously achieved; in addition, the invention has simple preparation and accurate, sensitive, simple, convenient and rapid detection and can detect a plurality of samples simultaneously.

Methods and compositions for high-throughput identification of protein / nucleic acid binding pairs are provided. In the subject methods, a nucleic acid probe array, e.g., a molecular beacon probe array, is contacted with a target nucleic acid population to produce a hybridized array. The resultant hybridized array is then contacted with a population of proteins to produce a protein bound array. Any resultant array surface bound target nucleic acid / protein complexes are then detected to identify protein / nucleic acid binding pairs. In certain embodiments, the protein and / or nucleic acid members of the identified protein / nucleic acid binding pairs are further characterized. Also provided are systems and kits for use in practicing the subject methods. The subject invention finds use in a variety of different applications.

The invention discloses a primer, a probe and a kit for fluorescence PCR (Polymerase Chain Reaction) detection of 18 types of high-risk human papilloma viruses. Different typing kits are detected by adding different specificity probes; the DNAs (Deoxyribose Nucleic Acids)) of 18 types of common high-risk human papilloma viruses internationally recognized and closely related to the cervical cancer can be detected once; and typing detection is carried out on HPV (Human Papilloma Virus)16 and 18. The application provides 6 universal primers and 18 specific molecular beacon probes. The DNAs of the 18 types of common high-risk human papilloma viruses can be amplified by the 6 universal primers; and meanwhile, the 18 probes are the specific molecular beacon probes designed by aiming at the 18 high-risk types; different types of probes are added according to the detection need and combined as the kits aiming at the detection need of the different high-risk types; and at the same time, the added probes are marked by different report genes, so that the purpose of carrying out typing detection on the high-risk HPV is achieved.

Encoded metal nanoparticles conjugated to oligonucleotides, and methods for their use are described.

A low cost benchtop molecular beacons (oligonucleotide probes) reader utilizes an array of light emitting diodes to excite the fluorescence of the reaction of the combining of the molecular beacons with actual DNA strands. If the Molecular beacons match with the DNA the reaction will fluoresce a known wavelengths. If there is no match, the reaction will not fluoresce. The molecular beacons reader uses an array of light emitting diodes at given wavelengths. The unit also incorporates the proper filters depending on the wavelength required. The unit has interchangeable cartridges with the filters and light emitting diodes that can be easily changed. The system is used after the PCR process (polymerase chain reaction) and the samples are viewed in PCR strips or PCR plates directly out of a thermocycler. This unit serves as a low cost go-no-go test with easy to use equipment.

The present invention provides novel quencher composition comprising anthraquinone quencher moieties. The anthraquinone quencher moieties are useful as quencher labels when attached to biomolecules such as natural or modified polynucleotides, oligonucleotides, nucleosides, nucleotides, carbohydrates and peptides. For example, polynucleotides can be labeled at the 3′ terminus with fluorescence quencher solid support compositions, and polynucleotides can be labeled at internally or at the 5′ terminus. The detectable probes may have a format like molecular beacons, scorpion probes, sunrise probes, conformationally assisted probes and TaqMan probes.

Methods of identifying molecular beacons in which a secondary structure prediction algorithm is employed to identify oligonucleotide sequences within a target gene having the requisite hairpin structure. Isolated oligonucleotides, molecular beacons prepared from those oligonucleotides, and their use are also disclosed.

The invention relates to an immunoliposome biochip, a preparation method thereof and an application thereof in biological detection. The immunoliposome provided by the invention is distributed on the chip in a micro-array way, and includes molecule probes, such as molecular beacon with fluorescence; and outside of the immunoliposome is loaded with various ligands. The immunoliposome biochip can specifically capture target cells in a cell mixture, and ensure messenger RNA and / or micromolecule RNA in the target cells without destroying biomarkers in the cells. In addition, the immunoliposome biochip provided by the invention can be used for capturing and selecting nanoparticles secreted by virus and cells, such as micro bubbles and exosomes. The immunoliposome biochip has advantages of simple operation, low cost, rapidness, sensitive and high selectivity, and can be used for molecular detection of various diseases, food safety and other fields.

Dual nucleic acid probes with resonance energy transfer moieties are provided. In particular, fluorescent or luminescent resonance energy transfer moieties are provided on hairpin stem-loop molecular beacon probes that hybridize sufficiently near each other on a subject nucleic acid, e.g. mRNA, to generate an observable interaction. The invention also provides lanthanide chelate luminescent resonance energy transfer moieties on linear and stem-loop probes that hybridize sufficiently near each other on a subject nucleic acid to generate an observable interaction. The invention thereby provides detectable signals for rapid, specific and sensitive hybridization determination in vivo. The probes are used in methods of detection of nucleic acid target hybridization for the identification and quantification of tissue and cell-specific gene expression levels, including response to external stimuli, such as drug candidates, and genetic variations associated with disease, such as cancer.

The invention discloses a transcription factor detection method based on DNA-Ag nanoclusters molecular beacons and exonuclease III cycle signal amplification. The novel nanomaterial DNA-Ag nanoclusters molecular beacon is applied as a fluorescent probe to transcription factor detection for the first time. Also, based on the characteristic that fluorescence adjustment of DNA-Ag nanoclusters molecular beacons is carried out by an enzyme cleavable DNA fragment, exonuclease III is added to participate in signal transformation and also participate in enhancement of the detection signal. The transcription factor detection method provided by the invention has the advantages of no label, high selectivity, low cost and high sensitivity, and can realize high throughput detection of transcription factors in biological samples.

The invention discloses a fluorescence chemical sensor and a method for simultaneously detecting diversified DNA (deoxyribonucleic acid) glycosylases on single-molecular levels and application of thefluorescence chemical sensor. The fluorescence chemical sensor, the method and the application have the advantages that the fluorescence chemical sensor is based on two different molecular beacons including a molecular beacon modified by 8-hydroxyl guanine and a molecular beacon modified by deoxygenated hypoxanthine, the tail end of the molecular beacon modified by the 8-hydroxyl guanine is labeled by cyanine 3 (Cy3) and quenching groups, the tail end of the molecular beacon modified by the deoxygenated hypoxanthine is labeled by cyanine 5 (Cy5) and quenching groups, the fluorescence chemicalsensor is used for detecting 8-hydroxyl guanine DNA glycosylases and N-methylpurine DNA glycosylases and is different from the traditional molecular beacons which can be severely affected by dynamicsand thermodynamics, signal restoration of the Cy3 and the Cy5 depends on molecular beacon splitting with the DNA glycosylases used as media, the DNA glycosylases can be simultaneously sensitively detected by the aid of the method without optional signal amplification, the activity of hOGG1 and hAAG can be detected by the aid of the method in an ultra-sensitive manner without optional signal amplification, the fluorescence chemical sensor can be easily, conveniently and quickly operated, and accurate and reliable test results can be obtained.

The invention discloses a detection kit for quickly identifying donkey skin, horse skin and mule skin, which is used for accurately identifying donkeys, horses, mules and jennets by creatively applying 16SrRNA genes of CKM (creatine kinase) karyogene (nDNA) and mitochondrial genome DNA (mtDNA) at the same time from genetic background of the horses, the donkeys and the mules. The kit adopts an molecular beacon probe (MB) method to carry out a quintuple multicolor fluorescence quantitative PCR (polymerase chain reaction) detection technology, and also can detect donkey, horse, mule and jennet components; and moreover, the kit is added with exogenous internal reference as an interior label, can detect and avoid false negative results produced by PCR inhibitor contained in a sample. The quintuple multicolor fluorescence quantitative PCR is used for detecting in the same tube without opening a cover, so that the detection kit is not easy to pollute, and is accurate and stable, simple to operate, extremely high in sensitivity, strong in specificity, and the like, and therefore, a novel way is explored for identifying the donkey skin, horse skin and mule skin.

The invention discloses a recombinase-mediated nucleic acid isothermal amplification reaction detecting method. The nucleic acid isothermal amplification reaction detecting method comprises the following step that 1, an amplification reaction system is prepared. The amplification reaction system mainly comprises a, a nucleic acid sample to be detected; b, a pair of oligonucleotides primer 1 and primer 2, wherein the primer 1 and the primer 2 are cross-fertilized with the nucleic acid sample; c, a fluorescent probe, wherein the fluorescent probe is one or multiple of a molecular beacon, a fluorescent resonator, a scorpion probe and a molecular torch, and the concentration of the fluorescent probe ranges from 50-900 nM / L; d, recombinase; e, DNA polymerase; f, single-stranded DNA binding protein; g, amplification reaction buffer. The invention further provides a detection kit based on the nucleic acid isothermal amplification reaction detecting method. According to the nucleic acid isothermal amplification reaction detecting method and the detection kit based on the nucleic acid isothermal amplification reaction detecting method, the complexity of the system is reduced, reagent cost is reduced, sequence synthesis of the probe can be achieved easily, the probe can be designed easily, and actual application and popularization are facilitated.

An assay and method of assay for optical detection of bioagents, a target nucleic acid or a target protein using a surface enhanced Raman scattering (SERS) active biomolecule molecular beacon. The present invention also provides the assay and method in a multiplexed format.

Provided herein is a library that comprises a plurality of molecular beacons (MBs), each MB having a detectable label, a detectable label blocker and a modifier group. The library is used in conjunction with nanopore unzipping-dependent sequencing of nucleic acids.

The invention relates to the field of microorganism detection, in particular to a staphylococcus aureus methicillin resistant strain PCR (Polymerase Chain Reaction) detection kit. The invention has the technical scheme of amplifying and detecting a highly conservative specific nucleic acid sequence nuc gene in the staphylococcus aureus gene by using the polymerase chain reaction and molecular beacon and fluorescent probe technologies and simultaneously amplifying and detecting the drug resistant nucleic acid sequence mecA gene to judge whether the strain is a methicillin resistant strain of staphylococcus aureus. The invention also provides PCR amplification primers of the nuc gene and the mecA gene and the sequence of the probe. The kit is applicable to the rapid detection of the staphylococcus aureus methicillin resistant strain in a clinical specimen of a suspect staphylococcus aureus infector and has the advantages of high sensitivity, high specificity, stability, timeliness, convenience for operation and the like.

This invention relates to a method of detection of food-borne pathogenic bacteria using multiple fluorescent PCR and modified molecular beacons. Several pairs of primers and modified molecular beacon probes are designed according to the specific gene sequence of the pathogenic bacteria to be tested, which is amplified using fluorescent PCR and modified molecular beacon. The amplified product is crossbred with molecular beacon, and the fluorescent intensity of the reaction system is tested with the necessary circle times for intended threshold as a judgment for results, that is, negative for Ct of 0 or 40 and positive for Ct less than 38. This invention, which is advanced in high sensibility, significant specificity, simple operation and intuitionistic observation, is applicable in simultaneous detection of any two kinds, three kinds and four kinds of randomly combined bacteria and food-borne pathogenic bacteria as well as specimen in large amounts.

The invention provides modified molecular beacons detectable by surface enhanced Raman spectroscopy (SERS) and related materials, processes, and methods of use. Examples methods provide for the determination of the presence or absence of a target nucleotide sequence in a sample nucleic acid by (a) providing a detection agent, which agent comprises: (i) a probe comprising a target complement sequence (TCS) being complementary to the target sequence and flanking the TCS, first and second oligonucleotide arms, said first and second oligonucleotide arms forming a stem duplex, and said first arm incorporating a first label moiety being detectable by SERS (e.g. a fluoroscein dye) and said second arm terminating in a second label moiety being detectable by SERS, which second arm further includes a surface seeking group (SSG-e.g. an azo-benzotriazole) capable of promoting association of the second label onto an enhancing surface ii) associated with said probe via said SSG, an enhancing surface, such that said first and second label moieties are in close proximity to each other and to the enhancing surface (b) exposing the sample to the detection agent, (c) detecting hybridisation of the TCS to any target sequence present in the sample by a change in the SERS spectra of said agent.

According to epidemic disease and electrical mirror morphological analysis to ill fish, the cause of disease is iris virus. Definite the iris virus ATPase gene conservative area sequence as augmentation target sequence, and design and synthesize double specificity primers and a molecule beacon probe to quantitative detect fish iris virus. According to the quantitative calibration curve made by different concentration gradient positive plasmid, the logarithm of different gradient quantitative mold number is correlation with Ct, and the correlation coefficient is 0.998; at the same time, the detection method has high sensibility, and it can detect at least 70 virus particles. Detecting other virus and LYCIV genes express that the method has good specificity.

The invention discloses an application of dual application reaction of Exo (exonuclease) III-assisted cycle and DNAzyme cycle to Hg<2+> detection. When Hg<2+> exists, T-rich parts of H-DNA (deoxyribonucleic acid) and A-DNA form a T-Hg<2+>-T metal base pair, and H-DNA and A-DNA are hybridized to initiate a chain replacement reaction to form a Hg<2+>-mediated double-chain structure (referred as to Hg-Complex) containing an overhung protection part and a DNAzyme part. Under the action of Exo III, Hg<2+> and A-DNA are released and are hybridized with another H-DNA for cycle to obtain a large amount of DNAzyme sequences. The obtained DNAzyme sequences can cyclically cut molecular beacons to generate amplified signals. By means of effective combination of two cycles, Hg<2+> in an aqueous solution can be detected sensitively by means of this strategy, and high selectivity is realized. In the meantime, the method can be used for detecting Hg<2+> in sewage successfully and provides a potential tool for quantitative detection of Hg<2+> in an environment water sample.

The invention relates to a surface-enhanced Raman technology based on signal-off and used for detecting intracellular telomerase activity. According to the technology, graphene, carbon nitride, MoS2, SiO2 and the like are taken as carriers, nano-gole or nano-silver with controllable dimension and morphology or a composite magnetic nano-particle with a core-shell structure is supported by the carriers, and then a hairpin probe containing a telomerase primer and a Raman molecule beacon is fixedly disposed on the nano=particle surface, so that the high-sensitivity high-selectivity SERS detection method based on signal-off is established. When a target exists, the primer extends and generates a DNA chain with a repeat sequence under the effect of telomerase, and the DNA chain is hybridized with the probe molecule, then the DNA hairpin structure is opened, a Raman signal molecule with the single-chain labeled end is far away from a Raman substrate, the Raman signal is reduced, and high-sensitivity detection on telomerase is realized. The probe prepared by taking Hela cervical carcinoma cell as a model realizes intracellular telomerase activity detection and dynamic detection on intracellular telomerase activity variation under effect of a telomerase inhibiting medicament. By using multiple cells for imaging, the probe is confirmed to be capable of distinguishing tumor cells and normal cells.

The invention relates to a multiple fluorescence PCR (polymerase chain reaction) kit capable of detecting five DNA (deoxyribonucleic acid) viruses of an aquatic animal simultaneously. The kit comprises a buffer solution, and also comprises five pairs of fluorescence PCR primers and five molecular beacon probes and a PCR primer, wherein the five pairs of fluorescence PCR primers and the five molecular beacon probes are respectively corresponding to channel catfish virus, epizootic haematopoietic necrosis virus, goldfish haematopoietic necrosis virus, fancy carp herpes virus and red porgy iridescent virus, the fluorescence PCR primers and the molecular beacon probes comprise nucleotide sequences shown in SEQ ID NO.3-17, and the PCR primer comprises nucleotide sequences shown in SEQ ID NO.1-2. The multiple fluorescence PCR kit capable of detecting five DNA viruses of the aquatic animal simultaneously can rapidly detect all the DNA viruses infecting the aquatic animals once, the sensitivity is increased, and the cost for detecting single sample is reduced.

The present invention provides a method for detecting fumonisin B1 based on fluorescence resonance energy transfer. The method comprises that: NaYF4:Yb,Ho upconversion fluorescence nanoparticles and gold nanoparticles are respectively and covalently bound to both ends of the stem-like part of the specific molecular beacon, wherein the fluorescence resonance energy transfer upconversion fluorescence can be quenched by the gold nanoparticles; and a fumonisin B1 aptamer and a complementary strand thereof are subjected to hybridization immobilization on the surface of magnetic nanoparticles, an analyte fumonisin B1 is added and is specifically bound with the aptamer so as to make the complementary strand be separated from the magnetic nanoparticles, and the separated complementary strand and the molecular beacon are subjected to hybridization so as to open the ring-like part of the molecular beacon, such that the fluorescence resonance energy transfer system is broken, the fluorescence signal is restored, and the fumonisin B1 content can be determined by detecting the fluorescence signal intensity, wherein the detection limitation of the method achieves 0.01 ng.mL<-1>. According to the present invention, the fumonisin B1 content in the food sample can be rapidly, accurately e and sensitively detected.