53results about How to "Increase fluorescence signal intensity" patented technology

The invention relates to a nucleic acid signal amplifying detection kit. The detection kit mainly comprises at least one first-stage signal amplifying probe and at least one second-stage signal amplifying probe or a third-stage signal amplifying probe aiming at each target gene, or mainly comprises at least one first-stage signal amplifying probe and at least one second-stage signal amplifying probe or a third-stage signal amplifying probe and a capture probe aiming at each target gene. The detection kit is designed by adopting a novel in-situ hybridization method, and fluorescent signal strength is improved through a signal amplifying system. The detection process can be completed within 8h, and a single-copied nucleic acid hybridization probe is combined with the corresponding fluorescent probe through the signal amplifying system, so that detection sensitivity of RNA in-situ hybridization is improved remarkably.

The invention relates to a digital signal processing method for detecting a microfluidic chip and an applied detection device. The microfluidic chip detection device comprises the microfluidic chip, a bifocal surface imaging fluorescence detection system, a signal processor, a display screen and a power supply, wherein a fluorescence signal of the microfluidic chip is received by the bifocal surface imaging fluorescence detection system and then is transmitted to the signal processor; the signal processor is provided with a digital signal processing display software system; and a derivative slope fit movement average space-time simple filtering algorithm software module, an image analysis and calculation module, a data query module and a storage display module are preset in the digital signal processing display software system to process data input by the bifocal surface imaging fluorescence detection system. Therefore, stepped distortion of a digital signal acquired by the invention is low; the background noise is low; and the sensitivity of biochemical analysis and molecular diagnosis nucleic acid amplification real-time fluorescence signal detection is effectively improved.

The invention relates to a nicking endonuclease-based netted rolling cycle amplification system and a use thereof. Based on the existing second-generation hyperbranched rolling cycle amplification technology, the amplification system utilizes a special endonuclease which is nicking endonuclease Nb.BsrDI so that a third-generation rolling cycle amplification technology which is a netted rolling cycle amplification (NGCA) technology is produced. Intensity of fluorescence signals produced by the netted rolling cycle amplification is increased with increasing of a target DNA concentration, is obviously stronger than that of fluorescence signals produced by linear rolling cycle amplification and hyperbranched rolling cycle amplification and has a detection limit of 0.1fM. Compared with the first-generation linear rolling cycle amplification and the second-generation hyperbranched rolling cycle amplification, the nicking endonuclease-based netted rolling cycle amplification system retains the prior art advantages in operationality, use cost and amplification time, further realizes signal amplification based on the prior art thereby providing good technical conditions for ultralow-abundance nucleic acid sample analysis and detection, and has a wide application prospect.

The invention belongs to a field of medicinal preparations, and relates to a liposome drug delivery system used for tumor imaging, wherein the system is modified by a polypeptide with an amino acid sequence of RPAKPAR and wraps a near-infrared fluorescent dye. According to the invention, the RPAKPAR polypeptide is a targeting head group and modified on surface of stealth liposome, and wraps the near-infrared fluorescent dye. After intravenous administration, through mediation effect of the RPAKPAR polypeptide, the carrier stealth liposome actively sends the near-infrared fluorescent dye to tumor positions, and penetrates tumor vasculatures into whole tumor tissues, thereby increasing distribution of the dye in the tumor positions; and then by using an in-vivo optical molecular imaging technology, the tumor imaging with relatively high fluorescent signal intensity can be provided.

The invention discloses a preparation of a detection chip, comprising the steps of: 1) designing oligonucleotide probe of bacteria; 2) preparing the chip. The invention further discloses a method for detecting pathogene in multiple food samples with the detection chip, comprising the following steps: 1) designing a primer; 2) extracting DNA from samples to be detected, and getting extracting solution; 3) multi-PCR amplifying of target gene; 4) hybridization detecting of chip; and 5) detecting and analyzing of hybridization result. The detection step of the invention is simple, easy to operate, and produces high correct rate result.

The invention discloses a detection kit for the expression of AR-V7. The kit includes a capture probe for detecting AR-V7 mRNA and a signal amplification system, and the signal amplification system includes an amplification probe and a labeled probe the tail end of which is modified by a fluorophore; wherein the capture probe is in a stem-loop shape and is used for connecting the target mRNA and the amplification probe; the amplification probe is used for connecting the capture probe and the labeled probe, the labeled probe is used for connecting the amplification probe and the fluorophore, each labeled probe has a P5 sequence which is complementary to and paired with the corresponding amplification probe P4 sequence, and the end is modified by the fluorophore. The detection kit has the advantages of high specificity, high signal to noise ratio, and high accuracy.

The invention discloses a fluorescent probe, a method for detecting tetracycline and an application of the fluorescent probe. The probe comprises a tetracycline monoclonal antibody and a signal carrier, wherein the signal carrier is an amino-functionalized aluminum nanosheet, and the particle size of the amino-functionalized aluminum nanosheet is 520-570nm. According to the invention, the probe isconstructed by a fluorescent metal organic framework material in immunochromatographic test strip detection for the first time, and a simple, novel, sensitive and rapid analysis system is developed for tetracycline detection of meat, milk, honey and other actual samples. The lowest detection limit of the provided test strip on tetracycline is 0.0516 ng / mL, and the value of the lowest detection limit is less than that reported by many other literatures.

The invention discloses an escherichia coli detection method based on a time-resolved fluorescence method and DNA hybridization. A capture probe DNA1 and an identifying probe DNA 2 are designed according to a DNA target sequence of escherichia coli; the capture probe DNA1 is fixedly marked on a glass sheet; and the identifying probe DNA 2 is jointed with long-life luminescent rare-earth europium complex. When the capture probe DNA1 and the identifying probe DNA 2 are connected in series and can generate base-pair complementary hybridization reaction with the target DNA (DNA of biology to be detected); and a DNA detection model based on two serial-connected probes on the surface of the glass sheet is established. The long-life luminescent rare-earth europium complex is used as a marker distinguishing the probe DNA 2; and the time-resolved fluorescence method can effectively eliminate the disturbance of fluorescent sources in a biological system and background light of a solid matrix. The detection method takes little time, is visual, flexible, accurate in reaction results and has very obvious advantages.

The invention discloses an osteosarcoma circulating tumor cell identification kit. The kit comprises capture probes for marker gene mRNAs and a signal amplification system. The marker genes comprise at least two epithelial cell marker genes selected from EpCAM, CDH1, KRT8, KRT16, KRT17, KRT18, KRT19 and KRT20 and at least two osteosarcoma metastasis-related interstitial cell marker genes selected from MMP2, MMP9, VIM, CDH2, FN1, HIF-1a, SERPINE1, SURVIVIN, TWIST1, AKT2 and SNAIL. The kit can completely detect osteosarcoma circulating tumor cells, evaluate their metastatic ability to avoid the possibility of the false negative result caused by different expression levels of CTCs in different types of cancers and further improve the accuracy of detection.

The invention relates to an ovarian cancer-related microRNA detection kit. In accordance with each to-be-detected ovarian cancer-related microRNA, the detection kit comprises at least one first-grade signal amplification probe, at least one second-grade signal amplification probe and at least one third-grade signal amplification probe, as well as a capture probe, wherein a P1 sequence of the capture probe of the ovarian cancer-related microRNA is selected from SEQ ID NO.1-SEQ ID NO.8. According to the detection kit provided by the invention, the selected capture probe and signal amplification probes, a hybridization reaction can be conducted under a uniform reaction condition, and non-specific binding can be avoided between the various probes; and the designed probes are good in specificity and high in signal-to-noise ratio in detection. Meanwhile, by combining the various probes, a system, which is complete in detection effect, is formed by the identifying kit and a detection method.

The invention discloses a method for improving fluorescence signal intensity of fluorescent protein. The method includes: using a signal peptide sequence of specific protein or a mutation sequence ofthe signal peptide sequence or chimeric sequence of the signal peptide sequence or a polypeptide sequence designed through artificial simulation to guide expression of the fluorescent protein so as toimprove the fluorescence signal intensity of the fluorescent protein in host cells, wherein the signal peptide sequence of the specific protein specifically refers to a signal peptide sequence of part of naturally existing protein or the mutation sequence of the signal peptide sequence or the chimeric sequence of the signal peptide sequence or the polypeptide sequence designed through artificialsimulation. By the method, the fluorescence signal intensity of the fluorescent protein in the host cells is improved, and fusion protein of this kind can be used as novel complete fluorescent proteinand also can serve as more efficient fluorescent indicating molecule to be used for various researches or research and development of fluorescence-labeled cells or organisms for other purposes.

The invention relates to a breast cancer related microRNA (Ribonucleic Acid) detection kit. The breast cancer related microRNA detection kit comprises at least one first-stage signal amplification probe, at least one second-stage signal amplification probe, at least one third-stage signal amplification probe and a capture probe which are provided for each kind of to-be-detected breast cancer related microRNA, wherein the P1 sequence of the capture probe of the breast cancer related microRNA is selected from SEQ ID NO. 1 to SEQ ID NO. 8. According to the breast cancer related microRNA detection kit disclosed by the invention, hybridization reaction can be carried out by the capture probe and the signal amplification probes which are selected by the breast cancer related microRNA detection kit under a uniform reaction condition, and all probes are not in nonspecific binding; the designed probes have good specificity and high signal-to-noise ratio during detection; meanwhile, a system with a good detection effect is formed by an identification kit and a detection method through combined use of multiple probes.

The invention provides a fluorescent dUTP-based method for automatically detecting an SSR molecular marker. The method comprises the following steps of: (1) preparing a PCR reaction system: mixing 1.0muL of 10*buffer, 25 to 50muM of dNTP, 2.0mM of MgCl2, 0.5muM of forward primer, 0.5muM of backward primer, 1 unit of Taq enzyme, 10pmol of fluorescent dUTP and 5ng of genome DNA, and adding super-pure water into the mixture to supplement the mixture to 10muL; (2) performing touchdown PCR: treating the mixture for 4 minutes at the temperature of 94 DEG C; performing 20 cycles for 30 seconds at the temperature of 94 DEG C, 30 seconds at the temperature of between 70 and 60 DEG C or between 66 and 56 DEG C and 1 minute at the temperature of 72 DEG C, wherein each cycle reduces 0.5 DEG Cm; performing 26 cycles for 30 seconds at the temperature of 94 DEG C, 30 seconds at the temperature of 60 DEG C or 56 DEG C and 1 minute at the temperature of 72 DEG C; and finally, treating the mixture for 1 minute at the temperature of 72 DEG C to obtain a PCR product; and (3) adding 9.34muL of super-pure formamide and 0.16muL of molecular weight internal mark into 1.0muL of PCR product, treating the mixture for 5 minutes at the temperature of 95 DEG C, then quickly cooling the mixture over ice, and performing mark detection on a sequencer.

The invention discloses a novel lipid material for realizing the delivery of a breast cancer targeted drug. The novel lipid material takes lysine as a connecting group and is connected with a cholesteric part, a biotin part and a glucose part. The affinity between biotin and glucose in the novel lipid material and biotin transporter (SMVT) and glucose transporter (GLUT1) can be utilized to realizethe double targeting function to breast cancer and play a stronger breast cancer targeting therapeutic role, wherein the biotin transporter (SMVT) and the glucose transporter (GLUT1) are highly expressed on the surface of breast cancer cells. The novel lipid material can be used in different dosage forms comprising liposomes, nanoparticles, micelles and the like, and the prepared paclitaxel-loaded liposome has obvious breast cancer targeting property and a wide application prospect.

The invention relates to a circulating gastric tumor cell identifying kit. The identifying kit, in accordance with each marker gene mRNA, comprises a capture probe, an amplification probe and a marking probe, wherein the marker gene mRNAs include three varieties: gastric tumor cell marker gene mRNA selected from at least one of CEA and HER2, epithelial cell marker gene selected from at least one of KRT7, KRT8, KRT17, KRT18, KRT19 and KRT20, and mesenchymal cell marker gene selected from at least one of AKT2, HIF-1A, TWIST1 and VIMENTIN. The identifying kit provided by the invention, which adds detection on gastric tumor cell marker gene on the basis of CTCs epithelial cell marker genes and mesenchymal cell marker genes which are common in various tumors, can avoid a false positive result caused by such factors as non-tumor epithelial cells in blood, the introduction of normal epithelial cells to a sampling process and the like, so that the detected epithelial cell marker gene and / or mesenchymal cell marker gene cells are circulating gastric tumor cells; and the accuracy and reliability of a detection result can be further enhanced.

Provided is a biochip and an apparatus for detecting a biomaterial. The biochip includes a metal thin film on the surface of a substrate, restraining autofluorescence of the substrate, and a spacer on the metal thin film, having capture molecules immobilized on the surface of the spacer and specifically bound to target molecules. The spacer has a thickness controlled to enhance the strength of a fluorescence signal emitted from a fluorophore labeled with the target molecules and immobilized on the spacer by the specific binding between the capture molecule and the target molecule.

The invention relates to a colorectal cancer early screening kit. The kit comprises capture probes, amplification probes and labeled probes for detection of target gene mRNAs. The target genes comprise colorectal cancer screening genes and non-blood cell related genes. The colorectal cancer screening genes comprise at least two of CK20, ZEB1, CEA and CD24. The non-blood cell related genes comprise at least two of EGFR, CDX2, MACC1 and LGR5. Through comprehensive assessment and statistical analysis and screening in lot of experiments, the colorectal cancer screening genes, the non-blood cell related genes and exclusion genes are selected so that detection of a single target gene is realized and simultaneous detection of multiple target genes is realized and thus the screening cells can be completely detected, leukocytes, hematopoietic stem cells and endothelial cells in blood are distinguished from the screening cells, false negative and false positive results are avoided and the accuracy of detection is further improved.

The invention provides a Twist gene expression detection reagent kit. The reagent kit comprises a false catching probe for detection of Twist gene mRNA, a catching probe and a signal amplification system, wherein the signal amplification system comprises an amplification probe and a labelling probe; the false catching probe is used for prehybridization of a biological sample, and the catching probe is a self-assembled polymerized and branched nucleic acid probe and is connected with target mRNA and the amplification probe, the amplification probe is used for being connected with the catching probe and the labelling probe, and the labelling probe is connected with the amplification probe and fluorophore. The reagent kit has the advantages of being high in sensitivity, high in specificity, high in accuracy rate and the like, and can be used for detecting the expression level of the Twist gene in the biological samples.

The invention relates to the technical field of endotoxin detection, and provides a fluorescent probe compound to solve a problem that fluorescent probes only can be used for detecting samples free from iodide ion displacement components. The fluorescent probe compound is 4-(4-(1,2,2-triphenylvinyl)phenyl)ethylpyridinium iodide, the molecular formula of the compound is C33H28NI, and the structuralformula is shown in the description. A method for detecting endotoxin by using the fluorescent probe compound comprises the following steps: (1) preparing a stock solution; (2) preparing a standard substance solution; (3) preparing a standard reaction solution and a reaction solution to be detected; (4) detecting to obtain a standard fluorescence curve and a fluorescence curve of the reaction solution to be detected; and (5) comparing the fluorescence curve of the reaction solution to be detected with the standard fluorescence curve to determine whether the endotoxin content in the reaction solution to be detected reaches a standard or not. When a system to be detected contains a component capable of displacing iodide ions, TPEPyE has a good water solubility, and cannot form an aggregatein a solution free from LPS molecules, so fluorescence cannot be emitted, thereby the interference of iodide ion substitute to detection is eliminated.

The invention discloses a method for improving the sensitivity of an immunochromatographic marker, which comprises the following steps of coupling an antibody with fluorescent microspheres to obtain an antibody-fluorescent microsphere conjugate, and coupling the antibody-fluorescent microsphere conjugate with the fluorescent microspheres again to obtain an enhanced antibody-fluorescent microsphereconjugate as the immunochromatographic marker. According to the method, the labeled antibody-fluorescent microsphere conjugate is coupled again, so that the same antibody can carry more fluorescent microspheres, the strength of fluorescent substances on a detection line is greatly improved, and the sensitivity of a reagent is improved. The invention also discloses an application of the method ininterleukin 6 detection.

The invention relates to an identification kit for circulating tumor cells of breast cancer. The identification kit comprises capturing probes, amplification probes and labeling probes aiming at marker gene mRNAs, wherein the marker gene mRNAs are selected from breast cancer cell marker gene mRNAs selected from at least two of HER2, SCGB2, MUC1 and MGB1, epithelial cell marker gene mRNAs selected from at least one of EpCAM, CDH1, KRT7, KRT8, KRT18 and KRT19, and mesenchymal cell marker gene mRNAs selected from at least one of FN1, CDH2, VIMNETIN, FOXC1 and TWIST1. With the identification kit, the false positive result caused by the factors including non-tumor epithelial cells in the blood and normal epithelial cells introduced in a sampling process is avoided, it is ensured that the cells in which the epithelial cell marker genes and / or mesenchymal cell marker genes are detected are circulating tumor cells of breast cancer, and thus the accuracy and reliability of the detection result are further improved.

The invention relates to a composite micro-nano structure array on a high light-transmission substrate and a method and application of the same. The invention is characterized in that micro-nano structure arrays of light guide materials on the high light-transmission substrate are taken as supporting structures of fluorescent conjugated polymers for detection. The method comprises: firstly, preparing the micro-nano structure arrays and secondary structure arrays of the light guide materials on the high light-transmission material substrate such as quartz and the like; and secondly, coating the fluorescent conjugated polymers which are sensitive to specified analyte on the surface of the nano structure substrate to form the composite micro-nano structure array which is used for detecting an object. The invention takes the micro-nano structure arrays of different light guide materials as adhesive substrates of the fluorescent conjugated polymers and utilizes the advantages of micro-nanostructures such as large specific surface area, evanescent wave effect, micro resonant cavity action and the like to improve the sensitivity of detection and the signal intensity, prolong the servicelife and improve the reutilization property. The composite micro-nano structure array on the high light-transmission substrate can be applied to the detection of the specificity and high sensitivity of solid, liquid and gas molecules.

The invention relates to a lung cancer early screening kit. The kit comprises capture probes, amplification probes and labeled probes for detection of target gene mRNAs. The target genes comprise lung cancer screening genes and non-blood cell related genes. The lung cancer screening genes comprise at least two of TTF-1, LUNX, NSE, EGFR, P63 and CEA. The non-blood cell related genes comprise at least two of CK7, CK19, hTERT, VIM, PIM-1 and Survivin. Through screening of target genes, detection of a single target gene is realized and simultaneous detection of multiple target genes is realized so that lung cancer early screening cells can be completely detected, leukocytes, hematopoietic stem cells and endothelial cells in blood are distinguished from the screening cells, false negative and false positive results are avoided and the accuracy of detection is further improved.

The invention provides a preparation method of a sequencing chip. The preparation method comprises the following steps of removing a natural oxidization layer of a surface to be processed of a chip, to obtain a bare chip surface, and then cleaning the bare chip surface; sequentially forming an oxidization layer and an HDMS layer on the bare chip surface; and through etching of photoresist, formingarray patterns on the HDMS layer, then performing etching for removing the exposed HDMS layer, performing deposition of an amino layer on a surface layer which is exposed after the HDMS layer is removed, and after the photoresist is removed, obtaining the sequencing chip. According to the method disclosed by the invention, the oxidization layer is re-generated for replacing the natural oxidization layer, so that the intensity of fluorescence signals can be effectively strengthened. Besides, through combination with the arrangement of a reflecting layer and metal grids, the intensity of the fluorescence signals is significantly strengthened when the chip is tested, and besides, signal crosstalk between adjacent pixels is isolated; and a sequenator using the chip is high in testing right rate, and is suitable for quick and efficient sequencing.

The invention discloses a novel parathyroid gland detection device which is composed of a probe, a handle, an external optical fiber cable and a host, a probe lens is arranged at the top end of the probe, the host comprises a cassette, an internal optical fiber cable, a laser emission unit, a fluorescence detection unit and a controller, and the external optical fiber cable and the internal optical fiber cable both adopt a single optical fiber design. The cassette comprises a laser light source, a photoelectric sensor, a first lens and a second lens, the laser light source is provided with a laser filtering cover, the surface of the photoelectric sensor is covered with a fluorescent filtering film, and the laser light source is placed on the photoelectric sensor. The parathyroid gland detection device is simple in structure, laser density can be improved, fluorescence signal intensity is enhanced, and accordingly parathyroid gland detection and recognition rate is improved.

The invention discloses a method for constructing a multifunctional upconversion nano-platform. The method includes loading antibodies or multifunctional human serum albumin on the surfaces of water-soluble upconversion nanoparticles through chemical bonding, and loading one or more fluoresceins on the surfaces of active groups such as the antibodies through hydrophobic interaction between proteins and fluorescein molecules so as to construct the multifunctional upconversion nano-platform. Compared with the prior art, the method has the advantages that a nano-composite prepared by the method is highly targeted, good in biocompatibility, simple in technological process, low in cost and easy to popularize and apply on a large scale, and the loaded fluorescein molecules are high and adjustable in concentration; the method has a promising application prospect in the field of biomedicine, tumor imaging and targeted delivering of anti-cancer drugs in particular.

The invention provides an InDel site genotyping method. The InDel site genotyping method comprises the following steps: (1) carrying out T vector clone sequencing on a DNA sequence of a targeted gene to obtain a targeted gene cloning product sequence, and carrying out multi-comparison on the targeted gene cloning product sequence to obtain an InDel site; and (2) designing a PCR amplification primer according to the InDel site, wherein the PCR amplification primer comprises a forward primer and a reverse primer, and a 5' tail end of the forward primer is modified by a fluorophore; and (3) carrying out PCR amplification on DNA of a sample to be detected by the PCR amplification primer to obtain an amplification product, and detecting the amplification product by fluorescent capillary gel electrophoresis to determine the genotype of the InDel site in the sample to be detected. The provided genotyping method is low in cost, simple and flexible to operate and accurate in result.

The invention relates to a protein typing kit of circulating tumor cells. The kit comprises capture antibodies against different CTCs marker proteins and labeled antibodies against different CTCs marker proteins. The CTCs marker proteins comprise epithelial cell marker proteins and interstitial cell marker proteins. The epithelial cell marker proteins are selected from EPCAM, E-cadherin, KRT5, KRT7, KRT-8, KRT-16, KRT-17, KRT-18, KRT-19, KRT-20. The interstitial cell marker proteins are selected from N-cadherin, vimentin, fibronectin, MMP2, MMP3, MMP9, AKT2, ZEB1, FOXC1, FOXC2, SNAI1, SNAI2, SERPINE1. The marker proteins selected in the invention can realize detection of single marker protein, and a single marker protein can be used with other marker proteins for the detection. Therefore, circulating tumor cells can be fully detected, and the cell type can be distinguished. And then, false negative results caused by difference of expression levels of some marker proteins among individual circulating tumor cells are avoided.