74 results about "Calcein" patented technology

The invention relates to an avian infectious bronchitis virus quick detection kit based on loop-mediated isothermal amplification technology and an application method thereof. The kit is composed of a (1) reaction solution, an (2) enzyme solution, a (3) primer solution and a (4) positive control solution, wherein the primer solution comprises three pairs of specific primers, i.e. inner primers, outer primers and loop primers. Before the reaction, a calcein-manganese complex is previously added, manganese is bonded with pyrophosphate ions precipitated by dNTP (deoxyribonucleotide triphosphate)to release calcein, and the result can be visually inspected and identified, wherein greenish yellow indicates a positive result, and light yellow indicates a negative result. The invention solves the problems of long period, low sensitivity, difficulty in field application and the like in the detection of avian infectious bronchitis virus in the prior art.

The invention discloses a preparation method of a layer-by-layer self-assembled double-modified liposome. The preparation method comprises following step: lecithin and cholesterol are taken as wall materials, and a nano liposome is prepared by thin-film dispersion-dynamic high pressure microfluidization method; and layer-by-layer self-assemble technique is employed for further treatment, wherein primary amino group on chitosan molecular chains is taken as positive charge, carboxyl on sodium alginate molecular chains is taken as negative charge, polyelectrolyte films are formed layer by layer by electrostatic interaction between the positive charge and the negative charge, and the polyelectrolyte films are assembled onto the surface of the nano liposome so as to obtain the sodium alginate and chitosan double-modified liposome. Average particle size of the double-modified liposome is 330.6+-37.3nm, zeta potential is -15.79+-0.697mV, and distribution coefficient is 0.65+-0.048. It is shown by the results of in vitro digestion experiment that in vitro digestion stability of the double-modified liposome is higher than that of unmodified liposome; and release rate of fluorescent substance calcein encapsuled by the double-modified liposome after 120min of digestion is just 38+-2%, and that of fluorescent substance calcein encapsuled by the unmodified liposome is 58+-2%.

The invention relates to a mycoplasma pneumoniae rapid detection kit and a use method thereof, wherein the kit holds a loop-mediated isothermal amplification reaction tube and BstDNA polymerase; the reaction tube holds reaction buffer, dNTP, magnesium sulfate, a primer 1:5-ACCAATGCCATCAACCCG-3, a primer 2:5-TACCGGCGTAACGCAAAG-3, a primer 3:5-ATTTTCACCCGTGAGGGGGAGTTTTCGCTTAACCCCGTGAACG-3, a primer4:5-ACAGCGCTAAGGGCATCACTGTTTTTCAAAGCCGCTTCGGTTC-3, lycine, manganese chloride and calcein. The method for detecting mycoplasma pneumoniae comprises the following steps: extraction of a sample to be detected or bacterium DNA to be detected, loop-mediated isothermal amplification reaction of mycoplasma pneumoniae and colour development detection. The invention has the characteristics of accurate detection, high sensitivity, strong specificity, simpleness, convenience and rapidness.

The invention discloses a reverse transcription loop-mediated isothermal amplification (RT-LAMP) visual kit for detecting Japanese B encephalitis virus and application of the kit. The visual kit contains a primer group and color development substances, wherein primers have sequences shown as SEQ ID NO.1-4, and the color development substances are calcein and manganese chloride. A using method of the kit comprises the following steps of: preparing an RT-LAMP system containing AMV reverse transcriptase, a 1 time reaction buffer solution, strand displacement DNA polymerase, a dNTP mixture, betaine, calcein, manganese chloride, MgSO4, a forward inner primer (FIP), a backward inner primer (BIP) group, an F3 primer, a B3 primer and RNA of a sample to be detected; performing thermostatic reaction on the reaction system at the temperature of between 61 and 65 DEG C, and inactivating; and judging a result under natural light or ultraviolet or natural light and ultraviolet, wherein if the reaction product is green under the natural light, the sample to be detected contains the Japanese B encephalitis virus; and if the reaction product represents obvious green fluorescence under the ultraviolet, the sample to be detected contains the Japanese B encephalitis virus.

The invention discloses a phytophthora capsici LAMP (Loop-mediated isothermal Amplification) primer and a rapid detection method thereof, which are specially used for phytophthora capsici specific detection. The invention mainly designs a phytophthora capsici LAMP primer (F3, B3, FIP and BIP); the primer is subjected to isothermal amplification and a 50micromole/L Calcein-500micromole/L MnCl2 developing agent is added to develop or agarose gel electrophoresis detection is carried out, so as to observe green fluorescent light or a ladder belt with an LAMP characteristic. The phytophthora capsici LAMP primer and the rapid detection method thereof can be used for rapid, sensitive and accurate detection of plants infected by phytophthora capsici and the phytophthora capsici in the soil in production practice and can also be used for early diagnosis of field diseases and the monitoring and identifying of bacteria, so as to provide reliable technological and theoretical foundations for preventing and treating the diseases caused by the phytophthora capsici.

The invention discloses a method for continuously determining the content of calcium oxide and magnesium oxide in bentonite. According to the method, a bentonite sample is added to a polytetrafluoroethylene plastic beaker, another beaker is not filled with the bentonite sample and serves as a blank control, then hydrochloric acid and nitric acid-hydrofluoric acid are added for dissolution respectively, perchloric acid is added for smoking, then hydrochloric acid is added to heat and dissolve salt, test solutions are transferred to large volumetric flasks respectively and diluted to a constant-volume scale, each solution is split into two parts of solutions with equal quantities; triethanolamine, water and hydroxylamine hydrochloride are added sequentially to one of the sample and the blank control, a potassium hydroxide solution and an appropriate amount of calcein are added, an EDTA (ethylene diamine tetraacetic acid) standard solution is adopted for titration until fluorescent green disappearance, the volume is recorded, and the content of calcium oxide is calculated; triethanolamine, water and hydroxylamine hydrochloride are added sequentially to the other one of the sample and the blank control, an ammonia-ammonium chloride buffer solution and an appropriate amount of eriochrome black T indicator are added, the EDTA standard solution is adopted for titration until purple red turns into blue, the volume is recorded, and the content of magnesium oxide is calculated.

The invention discloses a method for measuring the content of calcium oxide and magnesium oxide in refined slag and aims to provide a method which can be used for measuring the content of calcium oxide and magnesium oxide in the refined slag quickly, accurately and efficiently. A sample is dissolved with hydrochloric acid and nitric acid-hydrofluoric acid, perchloric acid is added, the mixture smokes and is taken off and cooled, hydrochloric acid is added along the wall of a beaker carefully, and salt is heated and dissolved; a solution is transferred to a large volumetric flask with secondary water, diluted until the solution reaches a predetermined scale and shaken uniformly, and two parts of the solution are taken out and put into two small beakers; triethanolamine, water and a little hydroxylammonium chloride are added to one part of the solution, a potassium hydroxide solution is added until the pH value of the mixture is 12, a proper amount of calcein is added, an EDTA (ethylene diamine tetraacetic acid) standard solution is dropwise added until fluorescent green disappears, and accordingly, the content of calcium is obtained; triethanolamine, water and hydroxylamine hydrochloride are added, an ammonium-ammonium chloride solution is added until the pH value of the mixture is 10, eriochrome black T is taken as an indicator, an EDTA standard solution is dropwise added until the mixture is in pure blue, and the content of calcium and the content of magnesium can be obtained.

The invention relates to a method for measuring the content of calcium ions in oil field water. Commonly, an EDTA (Ethylene Diamine Tetraacetic Acid) titration method is adopted to measure Ca2+ and an endpoint color is difficult to judge so as to cause inaccurate numerical reading. When the content of iron ions in water is over 1mg/L, the iron ions need to be removed, but the analysis step is cockamamie. Aiming at the measurement of the content of the calcium ions in the oil field water, a calcein indicator is used as an indicator; under the condition of a pH value of over 12, EDTA standard solution is adopted to titrate a water sample so as to measure the content of the calcium ions; the calcein indicator contains calcein, phenolphthalein and potassium chloride; the mass ratio of the calcein to the phenolphthalein to the potassium chloride is 20:7:2,000; and the EDTA standard solution has molar concentration of 0.0125mol/L. The measuring method disclosed by the invention is used for measuring the content of the calcium ions in the oil field water, is simple and convenient, has high accuracy and is suitable for in situ measurement. Moreover, the defects of difficulty in distinguishing a measurement endpoint and iron ion interference are overcome.

The invention discloses a primer, a kit and a method for detecting bacillus coli shiga toxin through PSR (Polymerase Spiral Reaction) isothermal amplification reaction detection. The primer comprisesa detection primer Ft and a detection primer Bt, wherein a nucleotide sequence of the detection primer Ft is shown in SEQ ID NO.1; a nucleotide sequence of the detection primer Bt is shown in SEQ ID NO.2. The invention also provides a kit for detecting the bacillus coli shiga toxin through the PSR isothermal amplification reaction. The kit comprises the primers, BstDNA polymerase and a mixed solution of calcein and manganese chloride; detection of polymerase spiral reaction on the bacillus coli shiga toxin can be carried out, time loss and short time consumption cannot be caused by the changeof a temperature; a reaction process is simple and convenient, a detection period is short, the specificity is strong, and detection results can be observed by naked eyes. According to the primer, thekit and the method for detecting the bacillus coli shiga toxin through the PSR isothermal amplification reaction detection disclosed by the invention, significance on amplification development of a novel constant-temperature amplification technology and detection on site of microorganism can be achieved.

The present document describes a screening composition comprising a marker compound, chosen from at least one of iodine, and fluorescein; eosin Y, erythrosine, ponceau S, calcein, a catalyst, chosen from at least one boron trioxide (B₂O₃), potassium (K), Gallium (III) oxide (Ga₂O₃), Nickel (II) oxide (NiO), Vanadium (V) oxide (V₂O₅), magnesium oxide (MgO), a bismuth oxide chosen from bismuth subcarbonate [Bi₂O₂(CO₃)], bismuth chloride oxide (BiClO), and bismuth oxide (Bi₂O₃), cesium bromide (CsBr), lanthanum (III) oxide (La₂O₃), molybdenum (VI) oxide (MoO₃), neodymium oxide (Nd₂O₃), Nickel (II) carbonate anhydrous (NiCO₃); and a pigment, chosen from at least one of scandium (III) oxide (Sc₂O₃), Lead (IV) oxide (PbO₂), Sulfur (S) powder, and Tungsten (VI) oxide (WO₃), chromium (III) oxide (Cr₂O₃), copper (II) oxide (CuO), copper (I) oxide (Cu₂O), iron (III) oxide (Fe₂O₃), lead (II) oxide (PbO). The document also describes method of using the same.

The invention discloses a loop-mediated isothermal amplication technology-based quick campylobacter jejunii detection kit and a using method thereof. The kit consists of (1) reaction solution, (2) a primer group, (3) sample pretreatment solution and (4) positive contrast solution, wherein the primer group is one of the following eight primer groups, and each primer group comprises a pair of primers, namely an internal primer and an external primer; and a calcein manganese complex is added in advance in the reaction, the manganese is combined with pyrophosphate radical ions separated by dNTP to release the calcein, the released calcein can be observed and identified by naked eyes, the positive result is yellow green, and the negative result is light yellow. The kit has the advantages of strong specificity, short reaction time, high sensitivity and the like.

The invention discloses a method for continuous determination of the content of Ca, Mg and Ba in Si-Ca-Ba-Mg alloy. The method includes the steps that a Si-Ca-Ba-Mg alloy sample is added into one beaker, no sample is added in the other breaker which serves as blank control, nitric acid-hydrofluoric acid mixed solutions are added respectively for dissolving the sample, and sulfuric acid is added till smoking happens; afterwards, hydrochloric acid is added for dissolving slats, water and hydroxylamine hydrochloride are added and heated to be boiled till the solutions are changed to be colorless from being yellow, the solutions are filtered into a large volumetric flask, sediments are washed by dilute sulphuric acid and water, and the sediments together with filter paper are placed into a porcelain crucible; ashing, firing and weighing are carried out, and accordingly the content of the Ba is obtained; two groups of quantitative solutions are selected to be put in beakers from the constant volume solution and the blank solution, one group is sequentially added with triethanolamine, water, hydroxylamine hydrochloride and a potassium hydroxide solution, several drips of magnesium sulfate are added, a proper quantity of calcein indicator is added, and EDTA is used for titration till fluorescent green disappears, and accordingly the content of the Ca is obtained; the other group is sequentially added with triethanolamine, water, hydroxylamine hydrochloride and an ammonia-ammonium chloride buffered solution, an eriochrome black T indicator is added, EDTA is used for titration till fuchsia is turned into blue, and accordingly the content of the Mg is obtained.

The invention discloses primers, kit and method for detecting Escherichia coli Shiga toxin I by PSR (polymerase spiral reaction). The primers for detecting Escherichia coli Shiga toxin I are designedaccording to specific target sequence stx1 of Shiga toxin I and include detection primer Ft and detection primer Bt, as well as acceleration primer IF and acceleration primer IB, and their nucleotidesequences are shown as SEQ ID NO. 1 to 4. The invention also provides a kit for detecting Escherichia coli Shiga toxin I by PSR; the kit includes the above primers, Bst DNA polymerase, and a mixed solution of calcein and manganese chloride, is suitable for detecting Escherichia coli Shiga toxin I by polymerase spiral reaction; after developing with a fluorescent dye, the results can be judged witheyes. The kit is simple and quick to operate, low in detection cost and suitable for field detection.

The invention discloses a calcein visual RT-LAMP kit and application thereof. The kit is characterized by containing a primer group, calcein, manganese chloride, a novel duck reovirus RNA template, RNase-free water, a 10-time reaction buffer solution, dNTPs, glycine betaine, MgSO4, Bst DNA polymerase and AMV reverse transcriptase, wherein the primer group comprises an upstream outer primer F3, a downstream outer primer B3, an upstream inner primer FIP, a downstream inner primer BIP, a loop primer LoopF and a loop primer LoopB, and the sequences of the primers are presented by SEQ ID NO: 1-6. According to the calcein visual RT-LAMP kit, the problems that a detection method for detecting the novel duck reovirus is complicated in operation, low in sensitivity and long in consumed time and cannot meet rapid field or base-layer detection requirements in the prior art are solved.

The invention discloses a marking method for calcein of sepiella maindroni otolith and a detection method thereof. The marking method comprises the following steps: adding calcein into aquaculture water, thereby preparing into a solution in concentration of 200-400mg/L; exposing to the sun; adding mianserin and uniformly mixing, thereby acquiring a marked soaking solution; hungering the to-be-marked sepiella maindroni and then soaking in the marked soaking solution; and soaking in fresh seawater after ending the soaking in the marked soaking solution, thereby acquiring the mark of the sepiella maindroni otolith. The invention has the beneficial effects that the marking method disclosed by the invention is simple, the purpose of nondestructive marking can be achieved and the large-scale release marking action can be realized at one time; the mixed compound with high-strength fluorescent light can be formed according to the marking method, the result observation and comparison become convenient and the marking time is long-lasting; the marking method is suitable for the sepiella maindroni in different stages; and an effective method is supplied for evaluating the proliferation release effect of the sepiella maindroni.

The invention discloses an LAMP detection primer, detection kit and detection method for 16S rRNA methylase rmtB. The detection primer disclosed by the invention consists of a pair of outer primers F3 and B3, a pair of inner primers FIP and BIP, and a ring primer L. The detection kit comprises a plurality of LAMP reaction tubes loaded with reaction liquid, and the reaction liquid contains 2* constant-temperature reaction buffer liquid, Bst DNA polymerase, primers and the like. The detection method comprises the following steps: firstly extracting DNA of bacterial genomes to be detected, adding the extracted DNA to the reaction liquid of the LAMP reaction tubes, performing a constant-temperature reaction in a water bath of 63 DEG C for 45-60 minutes, and judging the result according to the phenomenon whether white precipitate exists or not or whether the reaction liquid becomes green or not after calcein is added. According to the LAMP detection primer, detection kit and detection method disclosed by the invention, the problems that the conventional PCR technique is long in time, large in workload, large in cross contamination and complex in operation are solved, and the LAMP detection primer, the detection kit and the detection method have the advantages of strong specificity, high sensitivity and high rapidness, and low cost, are simple to operate, and are suitable for on-site rapid detection.

The invention discloses a continuous analysis method of calcium fluoride and silicon dioxide in fluorite. The method includes the steps that a fluorite sample is dissolved with glacial acetic acid, filtered by means of filter paper, precipitated and placed in a platinum crucible, and careful ashing and firing are conducted in a high-temperature furnace; residues are wetted by a few drops of water, hydrofluoric acid is added, low-temperature evaporation is conducted till the mixture is dry, then hydrofluoric acid is added again, the mixture is dried through low-temperature evaporation, and the product is taken out after firing and placed in a dryer, cooled and weighed; the platinum crucible is placed in a flask, hydrochloric acid is added for extraction, perchloric acid is added after evaporation, and the product is heated till smoking dry; the flask is taken down, hydrochloric acid dissolving salt is added, the test solution is transferred into a volumetric flask to be diluted for volume fixing, triethanolamine, water, hydroxylamine hydrochloride, a potassium hydroxide solution, magnesium sulfate and calcein are added, the mixture is titrated with an EDTA standard solution till fluorescent green disappears to become a finishing point, and the volume, consumed by calcium in the sample, of the EDTA standard solution is recorded; a blank experiment is conducted to obtain the content of calcium fluoride and silicon dioxide. The method has the advantages of being quick, accurate, simple in equipment requirement and low in analysis cost.

The invention relates to a method for marking grass carp otolith by fluorescent substances. According to the method, salt calcein is dissolved in water bodies, and marked solution of salt calcein solution is obtained; and fishes to be marked are put into the solution for soaking, and the marking of the fish otolith is completed. The salt calcein is adopted as the marking solution for soaking the fishes, the marking operation is completed, and the strong stress response damage and the fish body mechanical injury caused by artificial operation in the existing marking method are overcome; the investment of labor and materials is reduced; and the problem of high external hanging marker loss rate is solved. The method has the advantages that the cost performance is high, the operation is convenient, the soaking concentration and time can also be regulated according to different fishes, specifications and numbers, and the method belongs to a marking method capable of being applied to large-scale economic fish release.

The invention belongs to the technical field of metallurgical chemical detection and analysis, particularly relates to a method for detecting the calcium content in a solid-core calcium wire, develops a calcium content detection method which is simple and convenient to operate, low in cost, rapid, efficient and high in accuracy in the solid-core calcium wire, and establishes a method for determining the calcium content in the solid-core wire by using an EDTA (Ethylene Diamine Tetraacetic Acid) titration method. The dissolving effect of glacial acetic acid on calcium in the calcium line sample is fully utilized, so that the sample treatment is simpler and more effective; as hydrochloric acid is not added for treatment in the test, the interference of iron groups during titration can be effectively reduced; meanwhile, the calcein indicator with better pertinence is adopted, color change is clear, and end point judgment is facilitated; in addition, the outer wrapping iron sheet is cleaned, dried and weighed, then the content of each component is calculated, and the method is simple, visual, real, accurate and reliable in data. In general, the method is reasonable in process, safe, reliable, easy and convenient to operate, low in cost, rapid, efficient and capable of meeting the field fast-paced production requirement.

The invention discloses a measuring method of a calcium content in a silicon-calcium wire. The method comprises the steps of adding a silicon-calcium wire sample with mass of m into a polytetrafluoroethylene beaker, taking another polytetrafluoroethylene beaker without a sample as a blank control, then adding a nitric acid-hydrofluoric acid mixed solution to dissolve the sample, adding perchloric acid after the sample is dissolved, continuing to heat till sulfuric acid smokes, adding hydrochloric acid and heating to dissolve salt after no smoke emits and a test solution is dry and cooled, transferring the test solution to a high-capacity bottle, diluting to a scale for constant volume, taking a certain amount of solution from the solution after the constant volume into the beakers, sequentially adding triethanolamine, water and hydroxylamine hydrochloride, adding a potassium hydroxide solution to keep pH (potential of hydrogen) to be not less than 12, adding a plurality of drops of magnesium sulfate and an appropriate amount of calcein indicator, titrating with an EDTA (Ethylene Diamine Tetraacetic Acid) standard solution with a concentration of C till fluorescent green disappears as an end point, recording the volumes as V and V0, and obtaining the calcium content as follows: W<Ca>% is equal to C(V-V0)M<Ca>100/mK*100.

The invention relates to a method for preparing mesoporous silica nanoparticles loaded with calcein and wrapped by cationic liposome. The Calcein is loaded into mesoporous silica; the cationic liposome with uniform particle size is prepared to encapsulate the outside of the mesoporous silica nanoparticles loaded with the calcein. The cationic liposome prepared by the method encapsulates the calcein-loaded mesoporous silica nanoparticles, the prepared mesoporous silica nanoparticles loaded with calcein and wrapped by the cationic liposome have particle diameters of 80-130 microns, after cell addition, the cell survival rate is 80%-90%, the efficiency of viable cell labeling observed by an inverted fluorescence microscopy is about 90%-95%, the positions of viable cells can be judged by observing the distribution of the calcein in the nanoparticles, and high-efficient viable cell labeling is achieved.

The invention discloses a method for detecting S<2-> based on a calcein-rhodamine B-gold ion system. The method comprises the following steps: adding S<2-> solutions with different volumes into aqueous solutions of calcein-rhodamine B-gold ions; testing fluorescence intensity of the system under a condition that the excitation wavelength is 450nm; realizing detection on S<2-> by carrying out ionic quenching on fluorescence of the aqueous solutions of calcein-rhodamine B-gold ions through S<2->. The method is simple in operation and has low detection limit; moreover, the detection on S<2> can be quickly and accurately realized, and interference from other ions can be eliminated.

The invention discloses a determination method of the calcium content in a calcium containing ferroaluminium-manganese alloy. The method comprises the following steps of adding a calcium containing ferroaluminium-manganese alloy sample with the mass m into one beaker, enabling the other beaker to be empty so as to be used as blank control, later respectively adding hydrochloric acid, dissolving the sample on a low-temperature electric hot plate, taking out after the sample is dissolved, respectively and sequentially adding triethanolamine, water and hydroxylamine hydrochloride after cooling, adding potassium hydroxide solution to keep the pH not to be lower than 12, adding a few drops of magnesium sulfate and appropriate amount of calcein indicator, titrating by utilizing EDTA standard solution with the concentration C until the fluorescent green is disappeared as a terminal point, recording the volume as V and V0 respectively, and obtaining the calcium content as WCa% =C(V-V0)MCa100/m*100.