100 results about "Dual fluorescence" patented technology

The invention relates to a dual emission rate type fluorescent probe for visually detecting carbon dots-Au nanoclusters of mercury ions and a preparation method. Dual-emission composite silicon dioxide nanoparticles are composite silicon dioxide nanoparticles formed by utilizing carbon dot covered silicon dioxide particles as cores and covalently coupling the surfaces of the carbon dot covered silicon dioxide particles with the Au nanoclusters after surface amination. The carbon dots located in the silicon dioxide nanoparticle cores are taken as reference fluorescence signals, the Au nanoclusters on the outer layers are taken as response fluorescence signals, and the signals are used for Hg<2+> selective recognition. The Au nanoclusters as the response fluorescence signals are connected to the surface of a silicon layer through covalent bond connection, and one stable nano-fluorescent probe is formed. When the dual-fluorescence composite nanoparticles are taken as the rate type fluorescent probes, the intensity of the carbon dot fluorescence signals in the cores basically keep unchanged, and the Au nanoclusters on the outer layers can be bonded with Hg<2+> selectively so as to result in fluorescence quenching of the Au nanoclusters on the outer layers.

The invention provides a dual fluorescence quantitative RT-PCR (reverse transcription-polymerase chain reaction) specific primer for detecting type 1 and type 3 duck hepatitis A viruses. A dual fluorescence quantitative method comprises the following steps: comparing sequences of DHAV-1 and DHAV-3 published on GenBank, and designing a pair of specific detection primers SEQ1 and SEQ2 aiming at DHAV-1 and a Taqman probe (probe1) as well as a pair of specific detection primers SEQ3 and SEQ4 aiming at DHAV-3 and a Taqman probe (probe2) respectively in a region with conservation and relatively large difference between gene sequences of the two viruses; confirming the concentration of the dual fluorescence quantitative RT-PCR specific primer and the Taqman probe aiming at DHAV-1 and DHAV-3. The dual fluorescence quantitative method built by virtue of the group of primers is good in specificity and high in sensitivity, and can be used for quick serum type identification and real-time quantitative analysis of the type 1 and type 3 duck hepatitis A viruses.

The invention discloses a dual-fluorescence functional polymer nanometer microsphere with pH response and application thereof in tumor tissue detection and belongs to the technical field of high polymer materials. The method comprises the following steps: firstly synthesizing an emulsion microsphere of a composite fluorescence molecule A, introducing a polymer with a pH response function into a shell layer of the microsphere by adopting a seed emulsion polymerization method, and thus preparing the emulsion microsphere with a core-shell structure; further compounding a fluorescence molecule B into the shell layer of the microsphere, and thus obtaining the dual-fluorescence-emission functional polymer nanometer microsphere with pH response. The microsphere presents different fluorescence intensity and fluorescence color under different pH value conditions from acidity to alkalinity. The microsphere is applied to tissue detection based on the pH response function of the nanometer microsphere fluorescent property, the microsphere presents violet fluorescence in the normal tissue (pH about 7.4), the microsphere presents pink fluorescence in the tumor tissue (pH of less than 6.5), and the microsphere can be used for preparing a fluorescent material and has wide application prospects in the fields of tumor cell and tissue detection, drug sustained release and the like.

The invention discloses a dual fluorescence PCR detection reagent, a kit and a detection method for swine fever virus and African swine fever virus, and belongs to the field of animal pathogen detection. Aiming at a 5'-UTR gene of the swine fever virus and a P72 gene of the African swine fever virus, primers and probes capable of covering all strains and suitable for double detection are separately designed and screened, including two pairs of specific primers and two specific probes; the invention also describes a kit containing the primers and the probes and a PCR detection method using theprimers and the probes; the invention, through the design of the primers and the adjustment of each component of PCR, realizes purposes of one-time analysis and simultaneous detection and differentiation of the swine fever virus and the African swine fever virus on the premise of no reduction in sensitivity and specificity, which not only reduces the workload and cost of detection, but also greatly saves the detection time, thus gaining valuable time for epidemic disease prevention and treatment.

The invention belongs to the fields of chemistry, materials and food safety, and relates to a molecular imprinting ratio fluorescent sensor, a preparation method and application thereof. The preparation method of the molecular imprinting ratio fluorescent sensor includes the steps that molecular imprinting is carried out through sol-gel polymerization, single-component dual-fluorescence emission cadmium telluride / 8-hydroxyquinoline zinc nanoparticles provide fluorescence signals, holes after eluting bright blue are recognition sites, a mesoporous structure is formed after hexadecyl trimethyl ammonium bromide is eluted, and bright blue imprinted microspheres with the mesoporous structure are obtained, that is, the molecular imprinting ratio fluorescent sensor based on the single-component dual-fluorescence emission nanoparticles. According to the molecular imprinting ratio fluorescent sensor, preparation and application thereof, the obtained sensor prepared by the preparation method candetect bright blue with high selectivity and sensitivity, and provides color evolution and self-correction functions; and in addition, by effectively adjusting the emission wavelength of the sensor,the molecular imprinting ratio fluorescent sensor can be more widely used in detection of various colored substances.

Disclosed are activity evaluation of plant pathogens and a high throughput screening method for microbicides based on double fluorescent staining and a kit therefor, which include using a microtiter plate as tool carrier, using two different fluorescent dyes, respectively fluorescently labelling spores or mycelia of plant pathogens having vitality and no vitality, and due to the different colouring of spores or mycelia having no vitality and vitality, quantitatively detecting the survival rate of plant pathogens through fluorescence microscopy or flow cytometry, and accordingly preparing the kit. The method and kit can be used in aspects such as microbicide screening, evaluation of food safety, and water quality evaluation using the plant pathogen survival rate as an indicator. The method and kit are rapid, objective, simple and easy in operation, and have a low cost, and both can be used in studying the mechanism of the action of microbicides and high-throughput screening and the toxicity evaluation of microbicides.

The invention provides a nucleic acid dual fluorescence PCR (Polymerase Chain Reaction) detection kit for an influenza A / B virus. The detection kit comprises an RNA (Ribonucleic Acid) enzyme inhibitor, an RT-PCR (Reverse Transcription-Polymerase Chain Reaction) liquid, an enzyme mixed liquor, an influenza A / B virus dual reaction liquid, a positive control and a negative control, wherein the influenza A / B virus dual reaction liquid comprises a component (1) which consists of a pair of primers for detecting the influenza A virus and a probe for detecting the influenza A virus and a component (2) which consists of a pair of primers for detecting the influenza B virus and a probe for detecting the influenza B virus; and the enzyme mixed liquor comprises a Taq enzyme and a reverse transcription enzyme. By adopting the detection kit, the influenza A and B viruses can be detected at the same time, and the problem that only one influenza virus can be detected in one reaction in the conventional product is solved. The detection kit has the advantages of easiness and convenience for operating, high repeatability, quick and objective detection result and the like, and has a great application prospect in the field of in-vitro diagnosis of influenza viruses.

The invention belongs to the technical field of intelligent materials, and discloses a multi-stimulus-response organic small-molecular luminescent material, and preparation and application thereof. By utilizing the dual-fluorescence characteristics of phenothiazine derivatives, a series of luminescent materials are designed and synthesized; and external stimulus effects are utilized to regulate the reciprocal transformation of two conformations, thereby providing a simple feasible effective design strategy for the stimulus-response luminescent material. According to the preparation method, phenothiazine, which is used as a main reaction raw material, is subjected to a series of simple reactions to obtain various target products. The obtained product has the advantages of definite molecular weight, single structure, lower glass transition temperature and greatly higher stimulus-response sensitivity. The stimulus-response principle of the material is conversion between two different conformations instead of the influence of the intermolecular acting force and accumulation mode, so that the material has the multi-stimulus-response characteristic, and is applicable to sensors of force, temperature, pH and the like.

The invention discloses a preparation method and an application for a dual-fluorescence reporting system for detecting a micro-RNA (ribonucleic acid) function, wherein the preparation method comprises the following preparation steps of: A, obtaining a mCherry gene sequence via a PCR (polymerase chain reaction) amplification, inserting a rho EGFP-C1 carrier to replace the EGFP (enhanced green fluorescent protein) sequence of the mCherry gene sequence, and constructing a carrier rho mCherry-C1; B, using a RNAi-Ready rho SIREN-RetroQ plasmid as a template, obtaining a human U6 promoter gene via a PCR amplification, and inserting the carrier rho mCherry-C1 to obtain a plasmid rho hU6-mCherry-C1; and C, obtaining a gene sequence carried with a CMV (cytomegalovirus) promoter and an EGFP expression dialog sequence in an interval of 358 to 1944 on a plasmid rho Adtrack-CMV, and inserting the sequence in the plasmid rho hU6-mCherry-C1 to obtain a plasmid rho MGhU6. The plasmid rho MGhU6 is a dual-fluorescence reporting system simultaneously containing a mCherry reporting gene, an internal reference EGFP, a micro-RNA and the target sequence insertion site thereof. The reporting system can be used for detecting the inhibition function of the micro-RNA to the target sequence expression. The detection method is easy, as well as simple and convenient in operation; and a function detection for the micro-RNA can be realized by transfecting one plasmid only. The system also can be used for visually detecting the inhibition function of a micro-RNA in a single living cell to the target thereof by the aid of fluorescence microscopic imaging.

The invention provides a method for realizing detection of mercury ions in pure water solution in a ratio fluorescence form on the basis of a metal-organic framework material. According to the method, 5,5minute-(ethane-1,2-disubstituted bis(oxy)) m-phthalic acid is used as a ligand, and takes a hydrothermal reaction with hexahydrate cadmium perchlorate to obtain a three-dimensional metal-organic framework material; the material has dual fluorescence emission functions, i.e., a fluorescence emission peak based on the ligand and a charge transfer emission peak based on ligand-metal ions charge transfer are respectively included; the detection of the mercury ions by the metal-organic framework material in the ratio fluorescence form can be realized; the specificity on the mercury ion detection in alkali metal, alkaline-earth metal, transition metal and other heavy metal ions is shown; through much characterization, the mechanism of the material on the mercury ion detection is specific. Therefore the material provides a method for mercury ion ratio fluorescence detection on the basis of the metal-organic framework material, and the method has the advantages that the sensitivity is high, and the operation is simple.

The invention discloses a dual fluorescence quantitative PCR (polymerase chain reaction) detection method and a detection kit for clostridium difficile enterotoxin A and B. The method comprises the steps that 1) the DNA (deoxyribonucleic acid) of a sample to be detected is extracted; 2) the DNA (deoxyribonucleic acid) of the sample to be detected is used as a template and is subjected to fluorescence quantitative PCR reaction; 3) the fluorescence of each cyclic product in the PCR reaction is subjected to fluorescence detection, and whether the sample to be detected contains clostridium difficile enterotoxin A and B or not is judged according to the lowest Ct value and the highest fluorescence value in the fluorescence detection. The invention also discloses a specific primer, a fluorescence probe and the detection kit. The dual fluorescence quantitative PCR detection method is simple, convenient and quick to operate, can detect the clostridium difficile enterotoxin A and B simultaneously, is high in detection sensitivity and specificity and can be applicable to the laboratory emergency detection of outburst epidemic caused by clostridium difficile.

The invention discloses a dual-fluorescence staining solution for vaginal microorganism detection. The dual-fluorescence staining solution for the vaginal microorganism detection comprises an independent dual-fluorescence staining solution A and an independent dual-fluorescence staining solution B, wherein the dual-fluorescence staining solution A is composed of a fluorescence staining agent, an auxiliary staining agent, a staining solution buffer reagent, a bacteriostatic agent and water, and the dual-fluorescence staining solution B is composed of a staining solution buffer reagent, an alkaline regulating reagent, an anti-quenching agent, a bacteriostatic agent and water. The invention has the advantages of being simple in operation process, rapid in detection and easy to observe and recognize after pathogenic microorganism dyeing. Moreover, the interference of contaminated specimens such as bloody specimens, seminal fluid specimens and basal cell specimens on detection results is overcome, and the accuracy and the detection rate are effectively improved.

The invention relates to a reporter system for researching expression regulation of a Kiss1 gene and a construction method of the reporter system. The system is a dual-fluorescence Kiss1 gene expression regulation reporter system that red fluorescence protein achieves stable expression and green fluorescence protein is regulated by virtue of a Kiss1 expression regulatory element. The construction method comprises the following steps: constructing a red fluorescence protein targeting vector Ai9-dtomato; constructing a green fluorescence protein targeting system; constructing Cas9-GT1-7-Kiss1-2A-EGFP cells that EGFP is controlled by virtue of the Kiss1 expression regulatory element; and interpolating the cells into a red fluorescence background which is stable in expression, so as to obtain the reporter system. According to the reporter system and the construction method thereof provided by the invention, quantitative evaluation of a researched object on a Kiss1 action effect is achieved.

The present invention belongs to the fields of material, analytical chemistry and food safety, in particular to a molecularly imprinted ratiometric fluorescent nanosensor based on dual fluorescence emission, a preparation method and an application thereof in visual detection of folic acid in food. The surface of silica nanoparticles is subjected to imprinting polymerization with the sol-gel methodin one step; red fluorescent cadmium telluride quantum dots (CdTe QDs) and folic acid with spontaneous blue fluorescence are embedded; holes of eluted folic acid are used as recognition sites; then the dual fluorescence emission molecularly imprinted nanosensor with a core-shell structure is obtained, which is the visualized molecularly imprinted nanosensor. The preparation method of the invention is simpler than the conventional two-step method for preparing the molecularly imprinted ratiometric fluorescent sensor; cumbersome synthesis steps are avoided; and the experimental period is shortened. In addition, the sensor prepared by the method of the invention can be used for detecting folic acid with a high sensitivity, a high selectivity and self-correction; abundant fluorescence color evolution is provided; and the visual detection for a target is realized.

The invention belongs to the field of microbiological detection, relates to the detection of a PDCoV (porcine Delta coronavirus) and a PEDV (porcine epidemic diarrhea virus), and particularly relates to a dual fluorescence RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection method for the PDCoV and the PEDV and application thereof. The detection method comprises the following steps of designing upstream and downstream primers of the PDCoV and upstream and downstream primers of the PEDV; preparing a dual real-time fluorescence system; implementing a dual real-time fluorescence detection method. The invention also provides the application of the dual fluorescence RT-PCR detection method to the simultaneous detection of the PDCoV and the PEDV. The detection method established by the invention can be used for simultaneously detecting the two viruses of the PEDV and the PDCoV quickly and specifically, and has favorable specificity, repeatability and sensitivity; the strong technical support is provided for the clinical diagnosis and prevention and treatment of the PEDV and the PDCoV; a foundation is also laid for the molecular epidemiological and prevention and control research of such epidemic diseases.

The invention discloses a detection kit for detecting A-type H5N6 subtype avian influenza virus by utilizing dual fluorescence PCR (polymerase chain reaction). The detection kit comprises a PCR primer for amplifying H5 and N6 genes and a probe, the nucleotide sequence is shown in Seq ID No: 1-6, a fluorescence reporter group is marked at the 5' terminal of the probe, and a fluorescence quenching group is marked at 3 terminal; a reaction system comprises an RT-PCR reaction liquid, an enzyme mixing liquid, a positive standard substance and a negative standard substance. The invention further discloses a method for detection by utilizing the kit. The method comprises steps such as extraction of sample nucleic acid, preparation of a reaction system, amplification through fluorogenic quantitative PCR, result reading and the like. The detection kit can rapidly detect the A-type H5N6 subtype avian influenza virus and has the advantages of simplicity in operation, high sensitivity, good specificity and the like, and an efficient detection means can be provided for monitoring and clinical diagnosis of the A-type H5N6 subtype avian influenza virus.

The invention provides a dual-fluorescence PCR method for simultaneously detecting original components of horse and donkey, and belongs to the technical field of food detection. The method comprises the following steps: performing fluorescence quantitative PCR detection on a to-be-detected sample by using a primer of SEQ ID NO.1-6, judging whether the food contains original components of the horse and the donkey according to a Ct value. The detection method provided by the invention can fast identify and detect the original components of the horse and the donkey simultaneously within one reaction, the operation can be finished within 1-2 hours, and the method has the advantages of being accurate in detection, high in flexibility, strong in specificity, simple and fast, the lowest detection limit is 100fg, the requirements of fast identifying and detecting the original components of the horse and the donkey in large volumes can be satisfied, and the meat product adulteration can be effectively resisted, the protection on the consumer benefit is increased, and good social benefit is provided.

The invention belongs to the technical field of biology, and relates to a method and kit for detecting the copy number of CAR by dual fluorescence quantitative PCR. The primer pair for the method is selected from an arbitrary 1, 2 or 3 of the following 3 primer pairs: primer pair 1 which is a primer pair as shown in SEQ ID NO:1 and SEQ ID NO:2, a primer pair 2 which is a primer pair as shown in SEQ ID NO:4 and SEQ ID NO:5, and a primer pair 3 which is a primer pair as shown in SEQ ID NO:7 and SEQ ID NO:8. The method provides accurate technical support to quality control and curative effect monitor on CAR-T immunocyte treatment. The detection method can provide accuracy and sensitivity close to detection of single fluorescence quantitative PCR method, but dosage and operation for the methodis double saved, the experimental error caused by detection cost and operation can be remarkably reduced, and the detection efficiency and detection accuracy can be improved.

The invention discloses a dual-fluorescence quantitative PCR (Polymerase Chain Reaction) primer, kit and method for simultaneously detecting the general type and type 2 Streptococcus suis. The invention breaks through the limitation of the common PCR method, and can quantify the bacterial content in a sample in real time, has high sensitivity, strong specificity, good repeatability and obvious clinical diagnosis effects, the lowest detection limit reaches 5 copies per reaction; the Streptococcus suis and the type 2 Streptococcus suis can be specifically recognized, no response is generated to other bacteria; the variable coefficient of repeated detection is less than 1.19%; furthermore, the dual-fluorescence quantitative PCR primer for simultaneously detecting the general type and type 2 Streptococcus suis and the kit and method thereof disclosed by the invention are very suitable for detection of a large number of clinical samples and the routine epidemic surveillance of the Streptococcus suis, and extremely high in application values.

The invention discloses a mitochondria fluorescent probe as well as a preparation method and application thereof, belonging to the field of functional materials. The mitochondria fluorescent probe has a double fluorescence emission function and is a quaternary phosphonate compound with a structural formula shown by formula I. In the formula I, R is alkyl or aryl, and the anion X is halogen ion. The fluorescent probe disclosed by the invention has very efficient mitochondria specificity recognition ability, and the fluorescent probe molecule presents a double fluorescence emission phenomenon under the excitation of different lights so as to realize precise double fluorescence labeling of mitochondria. In the invention, the fluorescent probe molecule also has excellent light stability, experiences relatively little influence of the charge change on the mitochondria surface, and can realize real-time tracking of the mitochondria distribution and form in a living cell. Moreover, the mitochondria fluorescent probe disclosed by the invention has the advantages of high water solubility, simple synthesis path, simple after-treatment, batch production, relatively low cost and the like and shows a broad market prospect.

The invention belongs to the technical field of nano probes, and discloses a double-organelle-targeted nano probe as well as a preparation method and application thereof. The double-organelle-targetednano probe is mainly prepared from a compound of formula I and a photo-sensitive agent II; and the photo-sensitive agent II is phthalocyanines or porphyrins photo-sensitive agent with negative charges. The compound of formula I is a mitochondria-targeted chemotherapy reagent with an aggregation-induced luminescent effect. The nano probe of the invention not only can monitor the release process ina cell in real time by virtue of a double-fluorescent illumination way, but also can effectively improve the killing efficiency for cancer cells by virtue of the synergism of the mitochondria-targeted chemotherapy and lysosome-targeted photodynamics therapy. The invention also relates to application of nano probe in preparing an antitumor drug and / or fluorescent imaging. (The formulas are shown in the description).

The present invention discloses a dimer prodrug and a preparation method and an application thereof. A structural formula of the dimer prodrug is shown as follows. The dimer prodrug has a dual fluorescence effect, quenches fluorescences of CyNH2 and DOX through ICT and FRET effects, well monitors an activation process of a drug, also reduces toxicity, also utilizes a synergistic effect of the CyNH2 and DOX, and can also enhance a treatment effect on tumors.

The invention discloses a preparation method and application of dual-emission nitrogen-doped fluorescent carbon dots, the dual-emission nitrogen-doped fluorescent carbon dots are prepared by a solvothermal method, the fluorescent carbon dots have dual fluorescence emission wavelengths of 440 nm and 580 nm simultaneously under ultraviolet excitation, the fluorescence intensity proportion of the dual wavelengths is adjustable, and the dual-emission nitrogen-doped fluorescent carbon dots can be used for white light emission and temperature sensors. The fluorescent carbon dot disclosed by the invention has good fluorescence performance, particularly, the white carbon dot can achieve emission of pure white light (CIE coordinate 1931 (0.33, 0.33)), and is expected to be applied to the field of illumination. And the temperature range of 15-65 DEG C can be detected, and the temperature sensing performance is good.

The invention discloses a primer probe group for combined detection of Batai virus and Taina virus based on a dual fluorescence PCR method. The primer probe group contains specific primers and probesaiming at the Batai virus as well as primers and probes aiming at the Taina virus, wherein the specific primers aiming at the Batai virus include a forward primer and a reverse primer, the nucleotidesequence of the forward primer aiming at the Batai virus is represented by SEQ ID No.1, and the nucleotide sequence of the reverse primer aiming at the Batai virus is represented by SEQ ID No.2; and the specific primers aiming at the Taina virus include a forward primer and a reverse primer, the nucleotide sequence of the forward primer aiming at the Taina virus is represented by SEQ ID No.3, andthe nucleotide sequence of the reverse primer aiming at the Taina virus is represented by SEQ ID No.4. The primer probe group and the kit utilizing the primer probe group can be used for effectively detecting the Batai virus and the Taina virus, and the detection sensitivity can reach 1*10<3>copies / ml, so that the detection blanks of the Batai virus and the Taina virus in the prior art are effectively overcome, and the primer probe group and the kit have high industrial utilization values.

The invention discloses a dual-fluorescence plasmid carrying enhanced green fluorescent protein (EGFP) and red fluorescent protein (RFP) genes, wherein the EGFP and RFP genes are positioned in the same reading frame. The constructed dual-fluorescence plasmid can realize co-expression of two fluorescence proteins by placing two fluorescence protein genes in the same reading frame. Meanwhile, the dual-fluorescence plasmid is applied to deoxyribonucleic acid (DNA) mismatch repair functional activity analysis in a living cell to improve the verification accuracy at unicellular level, and the method is simple and has high sensitivity of indicating parameters and high accuracy. Therefore, the invention provides a novel effective detecting method for DNA mismatch repair functional activity detection in the living cell. The invention also provides application of the dual-fluorescence plasmid in preparing reagent for detecting the DNA mismatch repair functional activity in the living cell.