143 results about "Time resolved fluorescence immunoassay" patented technology

The invention belongs to the technical field of fluorescence immunoassay and relates to a photon counting multi-channel time-resolved fluorescence immunoassay system which comprises a light source module, a detection module and a time marked photon counting analysis/control module. At the same time, the invention relates to a counting method adopted by the system. The counting method of exciting a sample to be detected with pulse light, sending a fluorescence signal generated by the sample after excitation into a PMT (Photo Multiplier Tube) photon counting head and outputting a pulse signal comprises the following steps of taking cycles of the excitation pulse light as counting cycles, dividing each counting cycle into a plurality of time intervals with the same width, accumulating photons detected after sending excitation light pulses of each counting cycle into a memory corresponding to a corresponding time window, that is marking the positions of the photons in the pulse cycles with the intervals where the time windows are located, and uploading data in the memory into an upper computer after counting for the cycles. The multi-channel time-resolved fluorescence immunoassay system is low in cost and has very high sensitivity.

This invention relates to one agent case and its test method for corn zeranol, which is based on label immune reaction, wherein, the micro hole board comprises ZEN-BSA added with ZEN antibody; the free ZEN and micro board ZEN-BSA competes for ZEN antibody and the free ZEN antibody is removed with labeled sheep antipest antibody; the labeled immune reaction labeled antibody is removed; after adding strengthen liquid it uses time resolution fluorescence device to test the intensity cps with its concentration reverse to ZEN concentration to determine the ZEN content by comparing standard curve and sample.

The invention provides a kit for detecting diethylstilbertrol and detection method thereof, belonging the the time resolution fluoroimmunoassay (TRFIA) technical field, capable of detecting the content of diethylstilbestrol (DES) in animal-source food, blood, urine and feedstuff. The kit uses the TRFIA to detect the DES and the mark immune response is the base of the detection. A micropore plate is coated by DES-carrier protein and added with DES standard or sample and then added with DES antibody. The free DES and the DES-carrier protein on the micropore plate complete the DES antibody and the DES antibody which is not bonded with the free DES or the DES-carrier protein is cleared away and added with the EU3+-goat anti-rabbit antibody, after labeling the immune reaction, the non-bonded EU3+-goat anti-rabbit antibody is cleared away. After adding the reinforcement liquid, the fluorescence intensity cps is detected using a time resolution fluorescence instrument and the fluorescence intensity is inversely proportional to the DES concentration in sample and the content of DES in sample can be determined according to the standard curve. The DES detection kit has features of simple structure, convenient use, cheapness, high sensitivity up to 1ng/mL.

The invention belongs to the technical field of biology and relates to a protein chip reagent kit and a method for comprehensively detecting a lung cancer marker. The protein chip reagent kit for comprehensively detecting the lung cancer marker comprises: (1) a chip (1) and (2) a reaction agent and a detection agent, wherein a plurality of specific antibodies are simultaneously fixed on the chip, can generate antibody-antigen reaction with the lung cancer marker and are fixed on a bottom film of the chip to form a plurality of independent recognition sites; and the reaction agent and the detection agent are used for detecting whether a matter capable of generating antibody-antigen reaction with the specific antibodies exists in a sample to be detected or not through a TRFIA (Time Resolved Fluorescence Immunoassay) method. The reagent kit and the method have the beneficial effects that the six indexes of NSE (Neuron Specific Enolase), SCC (Squamous Cell Carcinoma), CEA (Carcino Embryonie Antigen), CA (Carbonic Anhydrase) 19-9, CYFR (Cytokeratin Fragment) A21-1 and pro-GRP (Glass Reinforced Plastic) can be simultaneously detected, the detection accuracy is improved, the repetitive experimental steps are reduced, and the time and the cost are saved.

The invention discloses a time-resolved fluorescence immunoassay reagent kit which is used for detecting quinoxalinone-2-carboxylic acid and a detection method of the time-resolved fluorescence immunoassay reagent kit. The reagent kit comprises a porous coated plate, a buffer solution, quinoxalinone-2-carboxylic acid standard, freeze-dried antibodies of quinoxalinone-2-carboxylic acid, europium-marked goat anti mouse antibodies, a cleaning solution and an enhancement solution. By measuring fluorescence intensity of counts per second (cps), the content of quinoxalinone-2-carboxylic acid in samples is calculated by a standard curve. The kit provided by the invention has the advantages of simple structure, easiness in assembly, corrosion resistance, light weight, low cost, wide application and the like. A valve core has good centering performance and can move flexibly and reliably and can play a good water-plugging role.

The invention relates to an analysis and detection technology of a human ovarian cancer tumor marker (human epididymis protein 4, HE4), in particular to a time-resolved fluorescence (TRF) immunized detection method and a kit of HE4, which is used for clinical auxiliary diagnosis, curative effect observation and prognosis judgment of the ovarian cancer. The invention comprises the following contents: constructing HE4 recombinant plasmids; expressing and purifying HE4 protein; immunizing BALB/c mice by the purified HE4 protein to prepare monoclonal antibodies; matching the obtained monoclonal antibodies to obtain two hybrid tumor cell strains (5A3 and 6C2) for secreting monoclonal antibodies of different epitopes of HE4 antigens, and marking the 5A3 monoclonal antibodies by the rare earth element Eu3+; and taking the unmarked 6C2 monoclonal antibodies as capture antibodies for coating a solid phase carrier, and taking the 5A3 monoclonal antibodies marked with the Eu3+ as detection antibodies to establish a double-antibody sandwich method for detecting HE4, thereby realizing the TRF immunized analysis of the invention.

The invention relates to the field of medical biotechnology, and particularly relates to anti-human tumor specific antigen ketoreductase 1B10 (AKR1B10) protein monoclonal antibody, and a time resolved fluorescence immunoassay (TRFIA) kit used for screening, diagnosis, efficacy judgment, prognosis evaluation or recurrence monitoring of cancers.

The invention discloses a double-labeling time-resolved fluorescence immunoassay method and kit for HIV (human immunodeficiency virus) antibody-HIV-1p24 antigen. The analytical method mainly comprises the steps of preparing a solid phase carrier coated with an HIV recombinant antigen and an HIV-1p24 monoclonal antibody simultaneously; preparing biotin-labeled HIV-1p24 monoclonal antibody; preparing lanthanide 1-labeled HIV recombinant antigen; preparing lanthanide 2-labeled streptavidin; adding a calibrator containing HIV standard antibody and HIV-1p24 standard antigen or a sample to be tested into the solid phase carrier coated with the antigen and the antibody, adding the biotin-labeled HIV-1p24 antibody, incubating, washing, then adding the lanthanide 1-labeled HIV antigen and the lanthanide 2-labeled streptavidin, incubating again, washing, and adding enhancement solution for fluorescence detection. The analytical method overcomes the difficulty that the antigen and the antibody cannot be distinguished in the existing joint detection for HIV antigen and antibody, realizes simultaneous and quantitative detection of the HIV antibody and the HIV-1p24 antigen and therefore shortens the window phase of HIV detection.

A detection method includes encapsulating microhole with OTA-BSA, adding OTA standard or sample and OTA antibody, washing out unconnected OTA antibody and EU3+ yangkangtu antiody, adding intensifier to determine fluorescent intensity CPS by time resolution fluorometer, presenting inverse ratio of fluorescent intensity to OTA concentration, comparing to standard curve for obtaining OTA concentration in sample. The kit applying TRFIA to detect OTA based on labelling immune reaction is also disclosed.

The invention discloses a time-resolved fluoroimmunoassay used for detecting Dkk1 and a kit thereof. The fluoroimmunoassay includes the following steps: 1. solid-phase anti-body preparing; 2) europium ion marker antibody preparing; and 3) the assay method: based on the double anti-body sandwiched immunoreaction. The invention has the advantages of relatively high sensitivity, specificity and stability on detection of Dkk1; the invention also has a supermatic analytical system which can not only enhance the speed of clinical test results, but also significantly reduce the personal errors and improve the reliability of the test results.

The invention discloses a time-resolved fluorescence immunoassay kit for detecting fumonisins B1 and a detection method thereof, belonging to the technical field of time-resolved fluoroimmunoassay (TRFIA) for detecting the content of FB1 in grain, fodder and food. The prepared kit adopts the TRFIA to detect the FB1. A micropore plate is coated with FB1-BSA, is added with FB1 standard or sample, and is added with FB1 monoclonal antibody. Dissociative FB1 and the FB1-BSA on the micropore plate compete for the FB1 monoclonal antibody, unconnected FB1 monoclonal antibody is removed by means of washing, Eu3+ -sheep anti-mouse antibody is added, and the unconnected Eu3+ -sheep anti-mouse antibody is removed by means of washing after labeled immune reaction. After adding strengthening liquid, fluorescence intensity (cps) is detected by a time-resolved fluoroimmunoassay apparatus, wherein the fluorescence intensity is in inverse proportion to the concentration of the FB1, and the content in the FB1 can be ensured by contrasting a standard curve. The kit has simple structure, convenient use, low price and high sensitivity more than 0.05ng/mL.

The invention discloses a time resolution fluorescence immunity analysis (TRFIA) method for synchronously detecting AFP and AFP-IgM and a reagent kit thereof. The reagent kit comprises the following reagents: 1) an AFP monoclonal antibody of a first epitope; 2) an AFP monoclonal antibody of a second epitope marked by a lanthanide element; 3) an IgM monoclonal antibody marked by another lanthanide element; 4) AFP/AFP-IgM mixed standard products; 5) a buffer solution; 6) washing liquid; and 7) enhancing liquid, wherein the lanthanide elements are selected from europium and samarium, and the AFP monoclonal antibody of the second epitope and the AFP monoclonal antibody of the first epitope have different epitopes. The invention adopts the TRFIA with the high sensitivity, the method for synchronously detecting AFP and AFP-IgM is established, the and the invention has the advantages of high sensitivity, strong specificity, good stability and simple operation, can realize the high atomization, can improve the speed of the clinical examination, can greatly reduce the artificial error, can improve the reliability of the detected results, and greatly improves the diagnosis efficiency.

The invention relates to a method for screening single chain antibodies of Microcystin-LR and verification thereof. The method comprises the following steps: carrying out two rounds of affinity screening on the biotinylated Microcystin-LR in a human source synthetic antibody library by using an avidin labeled magnetic bead and a negative screening method; extracting total plasmid DNA from phage colonies produced in the second round, carrying out enzyme digestion with enzyme Sfi I, recycling gel to obtain single chain antibody genes, connecting the single chain antibody genes with soluble expression carrier pAK100CL which is processed by enzyme digestion in the same way, and electrically transforming the connected carrier into colibacillus XL1-Blue to obtain soluble expression single chain antibodies; and verifying the soluble expression single chain antibodies by using a competitive time-resolved fluorescence immune analytical method. The invention has the advantage of quick, simple and convenient screening, and can well expose the Microcystin-LR three-dimensional structure into the incubation system. The verification on the screening result has the advantages of high detection signal and strong anti-interference capacity against stroma.

The invention discloses a time-resolved fluoroimmunoassay (TRFIA) kit and a detection method thereof for synchronously detecting the concentration of a free kappa light chain and a free lambda light chain. The kit comprises: 1) an antibody synchronously aiming at a heavy chain and a light chain of a human antibody, 2) an anti-free kappa light chain antibody labelled by lanthanide, 3) an anti-free lambda light chain antibody labelled by another lanthanide, 4) a standard substance of the free kappa light chain and a standard substance of the free lambda light chain, 5) a buffer, 6) a washing liquid, and 7) an enhancement solution. According to the invention, the antibody synchronously aiming at the heavy chain and the light chain of the human antibody is used as a capture antibody, other two antibodies respectively aiming at the free kappa light chain and the free lambda light chain are used as the labelled antibodies, double labeling TRFIA detection principles are used, thus the novel detection method for synchronously detecting the concentration of the free kappa light chain and the free lambda light chain is established. The method provided by the invention has the advantages of high sensitivity, strong specificity and good stability, and can realize high degrees of automation.

The invention discloses a human serum albumin (HSA) quantitative detection kit based on a time-resolved fluorescence immunoassay (TRFIA) technology. The kit comprises: a 96-pore microporous reaction plate coated with anti-HSA antibody, a calibrator, europium-labeled anti-HSA antibody, an assay buffer, a washing solution, and an enhancing solution. With a double-antibody sandwich immune reaction, a monoclonal antibody composition with coated anti-HSA antibody-HSA molecule-europium-labeled anti-HAS is formed; the enhancing solution is added, and value measuring is carried out by using a time resolution instrument. The method provided by the invention has the technical advantages of TRFIA, and can satisfy the requirement of quantitative detection of high-concentration (microgram-grade) HSA in biological product production. Therefore, the dilution of a product requiring detection during application is avoided, such that working time is greatly reduced, and working efficiency is greatly improved.

A reagent kit for detecting ochratoxin A and aflatoxin B1 at the same time and method thereof are provided, which belongs to fields of time-resolved fluoroimmunoassay. Micro plate is coated with OTA-BSA, AFB1-BSA, adding OTA, AFB1 standard or sample, and then adding OTA monoclonal antibody, AFB1 polyclonal antibody. Dissociated OTA, AFB1 and OTA-BSA, AFB1-BSA on micro plates competing corresponding antibody, and antibody which haven't be connected is eliminated after washing, adding Sm3+-goat anti rat and Eu3+- goat anti rabbit, labeled antibody haven't be connected after immune reaction are eliminated after washing. After adding enhancement solution, detecting Eu and scythe fluorescence intensity cps respectively which are inversely proportional to intensity of OTA and AFB1 in sample respectively by time resolved luminoscope, comparing standard curve and the content of OAT and AFB1 in sample is determined. The reagent kits of present invention has merits of simple structure, convenient operation and low cost, two examining results of OAT and AFB1are obtained by one operation, the present invention is used for detecting of grain, feed and food.

The invention relates to a homogeneous time resolved fluorescence immunoassay chelating agent and the preparation method thereof. The chelating agent is N, N, N', N'-((2, 6-di(3'-aminomethyl)-1-pyrazole)-4-(4-isothiocyanate(phenylacetylene)-pyridine))-tetraacetic acid. A luminous group is 2, 6-di(pyrazole) pyridine; the invention has high triplet-state energy level, the excitation wavelength after the combination with rare earth ions is 320nm, the emission wavelength is 520 to 620nm, the luminous service life is 2.99ms, the quantum yield is 0.58, the invention is in line with the requirements of homogeneous TRFIA to be combined with phenylacetylene at the pyridine 4-position, and the acetylene bond can block protein macromolecules from consuming luminous energy. The phenyl 4-linking isothiocyanate at the pyridine 4-position can be coupled with protein without injury, so as to be conductive to the high-efficient measurement of the luminescence of the chelating agent. The polyammonia polycarboxyl structure of 11 coordination site can exclude the quenching function of water molecules on rare earth luminescence and improve the solubility of the chelating agent in water and the dynamic stability of the chelating agent in the water r solution.

The invention discloses a method for detecting olaquindox by a directly-competing TRFIA (Time Resolved Fluorescence Immunoassay). The method comprises the following steps of: respectively adding standard olaquindox solution or treated sample solution and europium-labeled monoclonal antibodies into each micropore of an antigen precoating strip, mixing to be uniform, incubating for 1 hour at the temperature of 37 DEG C, and washing the micropores for 3-5 times by using washing liquid; adding 200muL of enhancement solution into each micropore, and carrying out light-proof oscillating incubation for 10 minutes at the temperature of 37 DEG C; determining the fluorescence intensity value cps by using a time resolution meter; drawing a standard curve; calculating out the corresponding concentration of the olaquindox from the standard curve according to the cpsx/cps0 value of each sample, multiplying by the corresponding dilution ratio and calculating out the actual concentration of the olaquindox in the sample. The method disclosed by the invention has the advantages that the operation is simple and convenient, the sensitivity is high, the stability is good and the lowest detection limit can reach 0.83ng.mL<-1>.

The invention provides a microfluidic time-resolved fluorescence immunoassay device. The device comprises a sample injection unit (101), a microfluidic control unit (103), a detection unit (104) and a microfluidic chip (102) adopting a membrane pump drive mode, and the microfluidic chip (102) is positioned in a microfluidic chip carrier on an analysis workbench of the microfluidic control unit (103). The microfluidic time-resolved fluorescence immunoassay device has the advantages that by adopting the microfluidic technology, a detection system is simplified; and a microplate oscillator and a microplate washer are omitted, a microfluidic control system is simple in structure, optimized, free of mechanical movements and structures of cleaning, vibration, transmission and the like, and automation is achieved conveniently.

The invention provides a lanthanide-series compound and a preparation method and an application thereof; the lanthanide-series compound has the chemical structural formula described in the description. The lanthanide-series compound has no inversion center, is beneficial for the compound to generate higher-brightness fluorescence, and enables a strongest emission peak to be used in a fluorescence analysis peak of time-resolved fluorescence immunoassay; and at the same time, the compound has relatively strong stability, and still can be used for marking and testing after being placed for a certain time.

The invention provides a kit for detecting chlorpromazine (CPZ) and a detecting method thereof, belonging to the technical field of time resolved fluoroimmunoassay (TRFIA) and used for detecting the content of CPZ in meat, fish and other animal derived foods. The kit prepared by the method is used for detecting CPZ by using TRFIA, and the detection is based on a labeled immunoreaction. A microporous plate is coated with CPZ-OVA (ovalbumin), a CPZ standard or sample is added, and a CPZ antibody is added; free CPZ competes for the CPZ antibody with the CPZ-OVA on the microporous plate, the CPZ antibody which is not connected is washed off and removed, Eu3+- antiMIgG is added, and the Eu3+- antiMIgG which is not connected is washed off and removed after the labeled immunoreaction; after an enhancing solution is added, the fluorescence intensity cps of the enhancing solution is detected by using a time-resolved fluorescence spectrofluorometer, the fluorescence intensity is in inverse proportion to the concentration of CPZ in the sample; and the content of CPZ in the detected sample can be determined in comparison with a standard curve. The kit for detecting CPZ provided by the invention has the advantages of simple structure, convenience in use, low cost and high sensitivity.

The invention provides a time resolution fluorescence immunoassay method based on magnetic separation; the method mainly comprises the following steps of firstly, coupling magnetic beads with an antibody to form immunomagnetic beads; meanwhile, enabling time resolution fluorescent microsphere to be coupled with antibody to form immunofluorescent microspheres; then enabling the immunomagnetic beadsand the immunofluorescent microspheres and the antigen in a sample to be oscillated and incubated in a reaction tube to form an immunomagnetic bead-antigen-immunofluorescent microsphere compound; andfinally, testing the fluorescence intensity, emitted by excitation of the compound at 360 nm excitation, by a time resolution instrument, wherein a standard curve is used as reference for determiningthe amount of the antigen in the sample. According to the analysis method, the reaction time is greatly shortened, and the detection efficiency and sensitivity are improved.

The invention relates to a test strip, a kit and a method for detecting AMH (anti-mullerian hormone) with TRFIA (time-resolved fluorescence immunoassay). The test strip comprises a PVC bottom plate, awhole blood filter pad, an antibody carrying membrane and absorbent paper. The kit comprises the test strip for detecting the AMH with the TRFIA, a sample buffer solution and an ID card containing anAMH standard curve. The kit can meet the requirement of single-person and small-batch detection of AMH and has the advantages of high sensitivity, high specificity, small blood amount, short detection time, operation convenience and detection result accuracy and reliability.

The aflatoxin B1 detecting kit and its detecting method belongs to the field of time-resolved fluorescent immunoassay (TRFIA) technology, and is used in the detection of aflatoxin B1 (AFB1) content in food and feed. The kit of the present invention detects AFB1 in TRFIA based on marker immune reaction. The microporous board has coated AFB1-HRP, added AFB1 standard or sample, and added AFB1 antibody. The free AFB1 and the AFB1-HRP in the microporous board compete AFB1 antibody, un-connected AFB1 antibody is washed off, EU3+ sheep anti-rabbit antibody is added and un-connected EU3+ sheep anti-rabbit antibody after marker immune reaction is washed off. After reinforcing solution is added, the fluorescence strength cps, which is proportional inversely to AFB1 concentration in the sample, is determined with time-resolved fluorescent instrument to determine the AFB1 content in the tested sample via comparison with standard curve.