60 results about "Shellfish poison" patented technology

The invention discloses a method for rapidly detecting shellfish toxin. The method comprises the following steps: (1) extracting and purifying shellfish toxin of a sample; (2) identifying the shellfish toxin; (3) preparing a shellfish toxin-BSA coupling antigen; (4) preparing and purifying a poisoning monoclonal antibody; (5) preparing immunomagnetic microspheres; (6) preparing a glucan-antibody-fluorescein mixed marker; (7) purifying the mixed marker; and (8) preparing a fluorescence immune chromatography test strip. The method is high in specificity and low in detection limit and can realize quantitative determination of antigen components.

The invention discloses a paralytic shellfish poisoning (PSP) standard sample and a preparation method and an application thereof. Positive shellfish raw materials of PSP are initially frozen and dried after being acidized and homogenized to obtain a crude sample, and then the crude sample is frozen and dried once again after being ground and sieved to obtain the product in the invention with good uniformity and stability. The standard sample can be used for capability assurance activities of PSP test items in labs, for PSP qualitative and quantitative detection and for verification of a detection method, correction of a tester and quality control and examination of test results. The invention has low raw material cost and simple preparation method, and the obtained standard sample is a material standard sample, is even and stable and easy for storage, and is beneficial for effective monitoring on shellfish poisoning.

This invention discloses a paralysis shellfish poison surface plasm resonance rapid measurement, which belongs to environment chemistry and biology molecule detecting field. The method is the following: to add quantitative PSP anti-body to the PSP sample solvent; then to flow on the gold sputtered plate with PSP solidified; to measure the over PSP anti-body in the solvent by use of plasm resonance sensor SPR detector to indirectly measure the concentration of the PSP solvent. This method is especially applied in quantitative study and regular or emergency monitor of PSP in freshwater algae or sea red tide.

The invention provides a method for measuring paralysis shellfish poison in aquatic products. The method comprises the steps of extraction, freezing, redissolving, purification and liquid chromatography-tandem mass spectrometer detection. The method has the advantages that sample pretreatment is simple, quick and economical, and sensitivity and recovery rate are high. The method is suitable for simultaneous, qualitative and quantitative detection of multiple kinds of paralysis shellfish poison in aquatic products, and provides effective technical support for guaranteeing quality safety of shellfish aquatic products in China.

The invention relates to a detection device for detecting red tide virus. An immune colloidal gold test strip for rapidly detecting paralytic shellfish poison disclosed herein comprises a sample pad, a gold jed pad, a Sartorius NC film, an absorbent pad and a PVC backing, wherein, the sample pad, the gold jed pad, the Sartorius NC film, and the absorbent pad are attached to the PVC backing orderly, the gold jed pad is coated with a colloidal gold-labeled paralytic shellfish poison monoclonal antibody, and the Sartorius NC film is respectively coated with a detection line consisting of Saxitoxin-bovine serum albumin conjugate and a quality control line consisting of goat-anti-mouse antibodies. The test strip has the advantages of strong specificity, high detection sensitivity, rapid detection, simple pre-treatment, no need of any device, convenience in carrying, low cost of the detection, simple operation, no need of professional personnel to operate, convenient storage, good stability, and high accuracy and high precision of the detection result, and the test strip can be stored for at least 6 months at room temperature.

The invention relates to batching detecting kit for oblivion bel toxin compete enzyme coupling immune that the feature is that it includes enzyme marking board, oblivion conch toxin standard antigen liquid, covering liquid, sealing liquid, horse radish peroxide enzyme labeling antigen bonder, substrate, drying out liquid and cleaning solution. It mainly makes the enzyme marking board. It is used to test the oblivion conch toxin and has high sensitivity, good specificity, high repeatability and rapid testing. It decreases testing cost, expands sample monitoring quantity and effectively controls biotoxin in food. It has important effect in rapid identifying, testing and curing in toxicant.

The invention discloses an ELISA method by indirect competition for testing the Gonyautoxin GTX2 and GTX3, which comprises the preparation method of antigen pre-peridium strips and the test method of Gonyautoxin. By utilizing the gained secretory positive cells of the monoclonal antibody with the function of anti-GTX2 and GTX3, the method provided in the invention can prepare in mass production the monoclonal antibody with the function of anti-GTX2 and GTX3, which costs low and can fast, conveniently and sensitively test the content of the GTX2 and GTX3 of the samples. The invention can make fast and sensitive test on the PSP (Paralytic shellfish poison) left in the seafood like fishes and shellfishes on a large scale.

The invention provides a removal method of paralytic shellfish poison, comprising the following steps: firstly, absorbent is distributed in embedding medium evenly; secondly, the embedding medium which contains the absorbent is solidified to form gel; thirdly, the solidified embedding medium which contains the absorbent is made into gel balls or blocks with uniform size; fourthly, the balls or the blocks are put in the solution which contains the paralytic shellfish poison, and the solution is then stirred for 5-30 minutes; and lastly, the gel is removed by filtration or centrifuge method to get the water which has removed the paralytic shellfish poison. The invention has the advantages that the absorbent which is treated by embedding can absorb the paralytic shellfish poison in the water. The absorption efficiency is higher than both pure absorbent and embedding medium. The embedded absorbent is comparatively big grain which is convenient to be separated from the system. The method can be used to remove the paralytic shellfish poison in the water, can be used for the enrichment and detection of positions, the purification of shellfish and so on, and can be used for the purification of other positions and pollutants in the environment after being improved.

The invention discloses a diarrhetic shellfish poison (DSP) standard sample as well as a preparation method and an application thereof. The method comprises the following steps: shelling fresh shellfish containing DSP masculine, taking out intestinal gland, adding organic solvent, homogenizing, freeze-drying primarily to obtain a coarse sample, grinding and screening the coarse sample to obtain freeze-dried powder, dispersing the freeze-dried powder in organic solvent again, and freeze-drying to obtain the DSP standard sample. The DSP standard sample has better uniformity and stability, can be used for proficiency testing activities in DSP testing projects in laboratories and for qualitative and quantitative detection of DSP, and can also be used for verification of detection methods, calibration of testing instruments and quality control and evaluation of testing results. The invention has the advantages that the raw materials cost is low, the preparation method is simple, and the obtained standard sample is a substantial standard sample, is uniform and stable, is easy to store, and is beneficial to the effective monitoring to shellfish poisons.

The invention discloses a DSP (diarrhetic shellfish poison) detection and analysis method based on a cell image sensor. The method is characterized by comprising the following steps: establishing the cell image sensor which is high in performance and low in cost, acquiring an image, converting an acquired RGB (red green blue) image to a grayscale image, binaryzating the image, matriculating the image, detecting a peak value, detecting the variation of a pulse image of a myocardial cell when in mechanical pulse, and detecting the mechanical pulse rate, pulse range and pulse interval of the myocardial cell; establishing a standard graph by detecting the response of the cell image sensor to different standard sample working solutions of the shellfish diarrhetic poisons in different concentrations; and detecting the response of the cell image sensor to the shellfish diarrhetic poisons of unknown concentration, comparing the detected response result with the standard graph, and semi-quantitatively calculating the concentration of the poisons. Compared with the existing shellfish diarrhetic poison detection and analysis method, the method is low in cost, simple in operation and capable of directly evaluating the toxicity of the poisons for a long time.

The invention discloses capillary electrophoresis electrochemical enzyme linked immunoassay for detecting amnesic shellfish toxicity, which belongs to the technical field of assay and detection. Solution of standard substances and shellfish samples with the amnesic shellfish toxicity are assayed by combining the capillary electrophoresis technology and electrochemical enzyme linked immunoassay technology. The capillary electrophoresis electrochemical enzyme linked immunoassay comprises the following steps of: reacting sample solution and an amnesic shellfish toxicity antibody marked by a horse radish peroxidase (HRP) in a noncompetitive mode; separating an amnesic shellfish toxicity-enzyme labeled antibody compound and the rest amnesic shellfish toxicity antibody marked by the HRP in the mixed solution by capillary electrophoresis; generating a 3-aminophenazine with electrochemical activities by using aminophenol which is the oxidogenic substrate of peroxide in the presence of a catalyst; and performing electrochemical detection. The method simplifies a sample treatment process and has high selectivity and high accuracy. The linear detection range of the solution of amnesic shellfish toxicity standard substances of the method is 0.5 to 50ng / mL and a detection limit of the method is 0.2ng / mL. The method is an ideal method for detecting the amnesic shellfish toxicity of shellfish samples.

The invention provides the rapid extraction procedures of the DSP in a shellfish sample aiming at the problems of complicated sample pretreatment, great workload, high cost and great time consumption in the existing liquid chromatography-mass spectrometry (LC-MS) technique for detecting a DSP (Digital Signal Processor). In the method provided by the invention, the UP LC / Q-TOFMS (Flight Mass Spectrometry) detection in the LC-MS technique are utilized and the UP LC / Q-TOFMS detection is used for the actual detection of merchant marine shellfish, thereby providing an effective and accurate detection technique for poisonous red tide research and food safety inspection.

The invention provides a method for purification preparation of N-sulfocarbamoyl toxins. The method comprises the steps of culture of toxin production algae, extraction of toxins, collection and purification of the toxins and the like. The process is simple and effective, meanwhile, the purity of the prepared N-sulfocarbamoyl gonyautoxins-2 (C1) and N-sulfocarbamoyl gonyautoxins-3 (C2) can reach higher than 91%, and the N-sulfocarbamoyl toxins can be used for quality safety detection and scientific research of saxitoxin in aquatic products.

The invention discloses a method for early detection on the toxicity of the Paralytic shellfish poisoning toxin in the technical field of the biological engineering, which is to make the check solution by the on-site shellfishes and make the abdominal cavity injection to the mouse with the check solution to test the content of the acetylcholine in the mouse brain and the change on activity of the endothelial nitric oxide synthase. The Paralytic shellfish poisoning toxin can be gained from the content of the acetylcholine and activity of the endothelial nitric oxide synthase, which can make an early detection on the toxicity of the Paralytic shellfish poisoning toxin. The invention realizes the on-site and fast test and analysis to large amount of samples and the early detection on the toxicity, which reduces the time by two to ten times compared with the common mice method currently used in China, avoiding any occurrence of the individual symptom and applicable to the on-site and fast test and analysis to large amount of samples and the early detection on the toxicity. The invention is simple and convenient to operate with only test on change of common biochemical parameters required, which is applicable to promotion and use by the on-site and grass-root test departments.

The invention relates to a detection technology for detecting paralytic shellfish poisoning, in particular to a method for preparing an anti-paralytic shellfish poisoning monoclonal antibody by using protein coupling to synthesize an antigen, and relates to the use of monoclonal antibody and gonylin GTX2 / 3 antigen Combination, and the preparation method of the quick detection kit that analyzes the content of gonylin GTX2 / 3 in the biological sample. An enzyme-linked immunoassay kit for rapid detection of the main component of paralytic shellfish poisoning, gonyellin GTX2 / 3, including a 96-well microtiter plate with detachable strips, enzyme-labeled secondary antibody, and buffer salt wash containing Tween-20 solution, substrate solution and stop solution, using the prepared monoclonal antibody containing specific anti-gonitoxin GTX2 / 3, on the 96-well ELISA plate coated with the coated antigen of the synthetic protein conjugate. The kit of the invention has the advantages of simple pretreatment, easy operation, and is suitable for on-site, fast, and large-volume sample detection. The kit has low cost and low price, and the detection limit of the kit is less than 15ng / ml.

The invention discloses a method for measuring diarrhetic shellfish poisons in shellfishes by use of immunoaffinity purification-liquid chromatography-tandem mass spectrometry. The method comprises the following steps: extracting a sample by use of an 80% methanol aqueous solution, mixing and diluting with a phosphate buffer solution, and performing immunoaffinity selective special purification, and performing liquid chromatography-tandem mass spectrometry. According to the use characteristics of a diarrhetic immunoaffinity column, parameters of sample liquid, eluate, eluent and the like are optimized; electrospray anions are ionized, a multi-reaction monitoring mode is adopted, and quantification based on an external standard method is achieved. The method is less in matrix interference, strong in purification effect, high in sensitivity and applicable to analysis measurement on diarrhetic shellfish poisons in shellfishes.

The invention relates to batching detecting kit for paralysis bel toxin compete enzyme coupling immune that the feature is that it includes enzyme marking board, paralysis conch toxin standard antigen liquid, covering liquid, sealing liquid, horse radish peroxide enzyme labeling antigen bonder, substrate, drying out liquid and cleaning solution. It mainly makes the enzyme marking board. It is used to test the paralysis conch toxin and has high sensitivity, good specificity, high repeatability and rapid testing. It decreases testing cost, expands sample monitoring quantity and effectively controls biotoxin in food. It has important effect in rapid identifying, testing and curing in toxicant.

An enzyme-liked immunodetection reagent box can rapid sensitively detecting red tide toxins Amnesic Shellfish Poisoning (ASP) in environmental sample. It utilizes synthetic high polymer ratio antigen DA-KLH, through many times immunity BLAB / c mouse, cell fusion, screening, induced ascites and protein column purification to obtain monoclonal antibody of idiocratic anti domoic acid (main constituent of Amnesic Shellfish Poisoning), selecting 1 : 1000 coat proportion and 1 : 8000 antibody dilution, developing out indirection competition enzyme-liked immunity analysing Amnesic Shellfish Poisoning (ASP) reagent box with sensitivity to 10 ng / ml.

The invention relates to the detection device of detecting the red tide toxin. An immune colloidal gold test paper to detect the lienteric toxin includes the sample pad, colloidal gold pad, NC film, the suction pad and PVC backing. The PVC backing is adhibited with the sample pad, colloidal gold pad, NC film and the suction pad. The colloidal gold pad is covered with the monoclonal antibody against okadaic acid marked by colloidal gold. The NC membrane is encrusted with the detection line composed by the okadaic acid and ovalbumin and quality controlling line composed by the multi antibody of sheep against rat. The test paper has high specialty, detection sensitivity, quick detection and simple pretreatment character. It needs not any device and convenience to carry and has low detection cost; the test paper has good stability and can be stored for six months in room temperature; the detection result has high veracity and precision.

The invention discloses a novel capillary electrophoresis electrochemical immunoassay for determining a diarrheic shellfish poison in a shellfish sample. After the diarrheic shellfish poison in the shellfish sample and an enzyme labeled diarrheic shellfish poison competitively react with limited antibodies, the diarrheic shellfish poison and the enzyme labeled diarrheic shellfish poison are separated in a capillary tube. The separated enzyme labeled diarrheic shellfish poison and the enzyme labeled diarrheic shellfish poison-antibody composite respectively react with substrates in reaction tubes and enter an electrochemical detection cell for end-column electrochemical detection. The content of diarrheic shellfish poison in the shellfish sample is calculated according to the drop of the peak of the enzyme labeled diarrheic shellfish poison-antibody composite. The invention has the advantages that the sample processing processes are simplified, the selectivity is good, the accuracy is high and the method is the ideal method for detecting the diarrheic shellfish poison in the shellfish sample.

The invention discloses a diarrhetic shellfish poison sample high-flux preprocessing device. The device comprises a control circuit, a switch power supply, a motor speed regulation plate, a peristaltic pump, a power supply interface, power supply sockets, a communication interface, a liquid tube interface, corresponding connecting tubes and a cabinet, wherein the power supply interface is connected with the switch power supply to input commercial power in order to supply power for the device; the power supply sockets are connected to the control circuit, and supply power for independent working devices; the communication interface is connected to the control circuit; the motor speed regulation plate and the control circuit are fixedly arranged above the switch power supply, and are connected with corresponding peristaltic pumps to control the working of the peristaltic pumps; the peristaltic pumps are fixedly arranged on the cabinet through a pedestal; the inner side of the front panel liquid tube interface is connected with the peristaltic pumps through Teflon tubes, and the interface is fixed through a band; and the outer side of the liquid tube interface leads out liquid flow through a Teflon tube. The device realizes high-flux preprocessing of a shellfish meat sample in the diarrhetic shellfish poison detection process, and improves the sample preprocessing efficiency.

The invention provides a method for preparing a diarrhetic shellfish poisoning okadaic acid monoclonal antibody. The method comprises the following steps: firstly, preparing an okadaic acid complete antigen; obtaining a hybridoma cell strain stably secreting an anti-OA (okadaic acid) antibody; finally preparing a large amount of monoclonal antibodies. According to the method provided by the invention, the preparation process is optimized, so that large-batch preparation of the diarrhetic shellfish poisoning okadaic acid monoclonal antibody becomes possible, and the foundation is provided for further developing an OA ELISA (enzyme-linked immuno sorbent assay) rapid detection kit which is high in specificity, free of cross reaction, sensitive and low in cost.

The invention discloses a method for simultaneously detecting AZA1, AZA2 and AZA3 in bivalve aquatic products. According to the method, liquid-liquid extraction and hybrid anion exchange reversed phase adsorption solid-phase extraction and purification are utilized to perform sample treatment on scallops, oysters, variegated clams and other bivalve aquatic products, and an analysis and detection method for simultaneously determining three main azaspiracid shellfish poisons AZA1, AZA2 and AZA3 in a sample through a liquid chromatography-tandem mass spectrometry (LC-MS / MS) method is established; and furthermore, the average recovery rate of AZAs is more than 80%, and the relative standard deviation (RSD) is less than 12%. The method disclosed by the invention has the characteristics of high speed, high efficiency, sensitivity and accuracy.

The invention relates to a method for preparing canned mactra veneriformis with primary taste, which comprises the steps of material selection, acceptance, selection, sand spitting, grading and selection, detection of mud clams, adding of concentrated clam soup and fine rice wine, vacuum sealing, sterilization and cooling, inspection under a constant temperature, boxing up and the like, specifically, selecting the mactra veneriformis of which heavy metal content, microbial indicators, saxitoxin and the like meeting relevant standards to carry out sand spitting and purification: wherein the water temperature is 20 DEG C to 30 DEG C, the salinity is 1.5% to 3.5%, and the time for spitting sand is 24 hours; then added 30g according to the proportion of 2:1 of the concentrated clam soup to the fine rice wine; packaging and sealing at a 700mm Hg vacuum level, wherein the sterilization formula is 5-5-5min / 125 DEG C; cooling down to a water temperature of 10 DEG C; and storing in a refrigerated warehouse after reaching commercial sterility through inspection. According to the method provided by the invention, the mactra veneriformis meat, not subjected to strong flavor conditioning but added with the special fine rice wine mixed with the primary taste soup, is processed into the special canned food with primary taste and has the advantages of simpleness in processing, convenience for eating and delicious taste, which provide a reference for promoting the utility value of mactra veneriformis and other low-value conches.

The invention relates to a preparation method and application of a neurotoxic shellfish poisoning immunoaffinity column, which belong to the technical field of shellfish poisoning detection. The preparation method of the neurotoxic shellfish poisoning immunoaffinity column is technically characterized in that the NSP immunoaffinity column is prepared by purifying a PbTX-2 monoclonal antibody and taking agarose gel as a carrier. According to the application of the neurotoxic shellfish poisoning immunoaffinity column in neurotoxic shellfish poisoning detection, cultured and sea-caught mussels, clams, oysters and scallops are taken as samples, the samples are extracted with an aqueous solution containing 80% of methanol and are diluted with a PBS buffer solution, and after purifying and enriching with the NSP immunoaffinity column, and LC-MS / MS test analysis is carried out. According to the method, PbTX-2 is taken as a research object, column packing of the NSP immunoaffinity column is self-developed and prepared, a sample extracting solution purified by the immunoaffinity column can be directly used for LC-MS / MS analysis, and the purposes of rapid detection and improvement of the sensitivity and accuracy of the detection method are achieved.

The invention belongs to the technical field of food safety inspection, and specifically relates to a diarrhetic shellfish poison immunoaffinity column, a preparation method and application thereof. The preparation method comprises the following steps: firstly using agarose gel activated by using an epoxy chloropropane activation method as a carrier, then coupling gene recombinant protein G onto the carrier to obtain protein G-sepharose, then connecting a diarrhetic shellfish poison antibody onto the protein G-sepharose so as to obtain antibody-protein G-sepharose, then using a cross-linking agent for cross-linking so as to obtain an antibody-protein G-sepharose filler, and finally packing the antibody-protein G-sepharose filler so as to form the diarrhetic shellfish poison immunoaffinitycolumn with high purity and affinity. The specific binding characteristic of the gene recombinant protein G and the diarrhetic shellfish poison antibody is sufficiently utilized, and the Fab segment of the diarrhetic shellfish poison antibody is exposed outside, so that the antibody combining capacity of the gene recombinant protein G, the capturing capacity of the diarrhetic shellfish poison andthe purification efficiency of the diarrhetic shellfish poison are greatly improved.

The invention discloses a method for detecting 13 saxitoxins in a biological sample. The method comprises the following steps of performing extraction treatment on a biological sample to obtain a sample extracting solution; by taking water as a solvent, respectively preparing standard substance solutions by using standard substances of 13 saxitoxins, namely C1, C2, GTX1, GTX2, GTX3, GTX4, GTX5, STX, NEO, dcSTX, dcNEO, dcGTX2 and dcGTX3; respectively preparing standard solutions by using the standard substance solutions of the 13 saxitoxins; respectively preparing a mixed standard working solution by using standard substance solutions of 13 saxitoxins; preparing a quality control sample; respectively sucking the sample extracting solution, the blank sample extracting solution, the added sample extracting solution, the standard solution and the mixed standard working solution, detecting by adopting a liquid chromatography-mass spectrometer, analyzing and calculating a detection result. The method has advantages of being simple in determination method, easy to operate, good in repeatability, high in sensitivity and the like, is suitable for the field of forensic science, and realizes high-sensitivity simultaneous determination of multiple saxitoxins in a biological sample.

The invention discloses malic acid-chitosan nanopore hydrogel microspheres as well as a preparation method and application thereof, and belongs to the technical field of guaranteeing the safety of aquatic food and preparing paralytic shellfish toxin bio-adsorbents. The invention discloses a preparation method of malic acid-chitosan nanopore hydrogel microspheres. The preparation method specifically comprises the following steps: preparing malic acid and chitosan into hydrogel; preparing a malic acid-chitosan nano-pore hydrogel microsphere, adding nano-silica and glycerin into the prepared hydrogel, adding sodium hydroxide, completely dissolving the nano-silica under an alkaline condition to form uniformly distributed nano-pores, performing washing, performing freeze-drying, and performing grinding and sieving to obtain the malic acid-chitosan nano-pore hydrogel microsphere. The malic acid-chitosan nanopore hydrogel microspheres prepared by the invention can be used as an efficient adsorbent for adsorbing and removing paralytic saxitoxin in a water body. The preparation method disclosed by the invention is simple, convenient to use and easy to store after being dried, and has great application significance on shellfish toxin pollution and improvement of product safety.

The invention belongs to the technical field of mass spectrometric detection, and discloses a high-resolution mass spectrometric detection method for lipophilic toxins in shellfish. The method comprises sample treatment and qualitative and quantitative detection, and the sample treatment method comprises the following steps: weighing a sample, putting the sample into a centrifuge tube, adding methanol, carrying out vortex uniform mixing, carrying out ultrasonic extraction, carrying out centrifuging, taking the supernatant, and transferring the supernatant into another centrifuge tube; repeatedly extracting residues once by using methanol, merging the supernatant, fixing the volume by using methanol, adding anhydrous magnesium sulfate and C18, violently performing oscillating, then carryingout centrifuging, and filtering the supernatant by using a filter membrane. The qualitative and quantitative detection comprises the step of performing determination by using a liquid chromatography-quadrupole rod-electrostatic orbitrap high-resolution mass spectrum after the sample is treated. The method can be used for qualitatively and quantitatively analyzing various lipophilic shellfish toxins at the same time, and has important reference significance for knowing the pollution condition of the shellfish toxins.