57 results about "Pepsinogen II" patented technology

The invention discloses a latex enhanced turbidimetric immunoassay kit for diagnosing gastric diseases or gastric cancer, a preparation method thereof and application. Components of the kit include diluent, blank solution and a latex reagent of an antibody of pepsinogen I or an antibody of pepsinogen II and can further include a calibrator and quality control serum. The latex reagent contains nanoparticles coupled with the antibody of the pepsinogen I or the antibody of the pepsinogen II, and the particle sizes of the nanoparticles are different. The examination sensitivity cannot meet requirements when latex with the single particle size lower than 100nm is used for examination, the linear range is small when latex with the single particle size of about 200nm is used for examination, and accordingly the examination sensitivity and the linear range cannot meet requirements when latex with the single particle size is used for examination. After composite latex of the kit is used for marking, the examination sensitivity can be improved, the linear examination range can be broadened, and the latex enhanced turbidimetric immunoassay kit has the advantages of fast examination, high sensitivity and specificity, good accuracy and the like in terms of gastric disease or gastric cancer examination.

The invention discloses an enzyme linked immunosorbent assay kit for combined diagnosis of the gastrosis or evaluation of gastric cancer risks and a preparation method thereof. The kit comprises a micropore plate coated with an antibody against a pepsin antigen I or an antibody against a pepsin antigen II, an enzyme labeled antibody, a color-developing agent, a stop solution and a concentrated cleaning solution, wherein the pepsin antigen I or the pepsin antigen II is a natural protein obtained from extraction of human gastric mucosa tissue. The kit disclosed by the invention adopts a mouse immunized with pepsinogen I and pepsinogen II which are separated from human gastric mucosa to prepare immunogen of a monoclonal antibody, the used standard sample also adopts the pepsin antigen I or the pepsin antigen II separated from the human gastric mucosa, thereby the defects caused by adopting different structures of animal pepsinogen and human pepsinogen are filled. The kit can be used for accurately diagnosing the gastrosis or early gastric cancer and has the advantages of high sensitivity, strong specificity, good accuracy and the like.

The invention relates to the technical field of medical examination and particularly relates to a latex immunoturbidimetry type pepsinogen II detection kit capable of eliminating chyle interference. The kit provided by the invention comprises: (1) a pepsinogen II calibrator; (2) a reagent 1 with a preset chyle remover; and (3) a reagent 2 containing a monoclonal antibody of anti-human pepsinogen II and polyclonal antibody coated latex particles. According to the kit using the method, a conjugation reaction of substance to be detected in a sample and specific antibodies in the reagents is amplified by a latex agglutination effect, turbidity formed by the reaction is associated with the content of the substance to be detected under a given wavelength, so that the content of the substance to be detected can be calculated. The kit provided by the invention is used for detecting the content of pepsinogen II in human serum, and the kit is high in sensitivity, good in specificity, capable of eliminating chyle interference in the sample, quick and convenient to operate, strong in practicability and is wide in application range.

The invention discloses a recombinant human pepsinogen II isozyme chimeric protein, and a preparation method and applications thereof. The preparation method comprises the steps as follows: by taking gene sequences of two isozymes of recombinant human pepsinogen II as a template, carrying out PCR (polymerase chain reaction) splicing to obtain a chimeric protein coding gene sequence, constructing recombinant expression plasmids, transforming the screened positive clone plasmids into expression host cells, screening an efficiently-expressed recombinant chimeric protein strain, carrying out enlargement culture on the cells and inducing expression chimeric protein, and purifying to obtain the recombinant human pepsinogen II isozyme chimeric protein. The invention further discloses the applications of the recombinant human pepsinogen II isozyme chimeric protein in preparing monoclonal antibodies and multiresistant serums or pepsinogen II kit calibration products. A great amount of stably expressed recombinant human pepsinogen II isozyme chimeric protein can be produced by utilizing a genetic engineering technology, and one protein has two chimeric protein sequences of the human pepsinogen II.

The invention relates to the field of immunodetection, in particular to a chemiluminescent quantitative determination kit for pepsinogen II and a preparation method of the chemiluminescent quantitative determination kit. The chemiluminescent quantitative determination kit for the pepsinogen II comprises (1) a pepsinogen II antibody coated micropore plate, (2) an HRP-labeled pepsinogen II antibody, (3) series pepsinogen II calibration materials diluted by a calibration material diluent, (4) a luminescent substrate A, (5) a luminescent substrate B and (6) a solid washing liquid. The kit disclosed by the invention is capable of detecting the content of the pepsinogen II in serum; and the level of the pepsinogen II in the serum is in positively relevant to atrophic gastritis and peptic ulcer; in treatment of peptic ulcer, the treatment result can be judged by monitoring the change of the degermed pepsinogen; and the chemiluminescent quantitative determination kit is noninvasive, simple and fast in detection method, high in sensitivity, wide in detection range and the like.

The invention discloses a kit for detecting pepsinogen II, belonging to the technical field of biological detection. The kit comprises a reagent R1 and a reagent R2, wherein the reagent R1 comprises components of the following concentrations: buffer of 10-200mmoL / L, surfactant of 0.01-5g / L, accelerator of 5-20g / L, and preservative of 0.05-2g / L, wherein a component concentration in the reagent R1 is the final concentration of a component in the reagent R1; reagent R2 comprises the components of the following concentrations: buffer of 80-160mmoL/L, stabilizer of 10-120g/L, blocking agent5-30g/L, anti-human pepsinogen II antibody latex solution of 0.5-3.0g/L, and preservative of 0.05-2g /L, wherein a component concentration in the reagent R2 is the final concentration of a componentin the reagent R2; the stabilizer is at least one of glycerin, sucrose, trehalose and glucose. The kit of the invention has the advantages of high sensitivity, high precision, stable reagents and thelike.

The invention discloses a pepsinogen II detection kit. The kit comprises a magnetic particle coated with a pepsinogen II monoclonal antibody, an enzyme diluent containing a horseradish peroxidase (HRP) labeled pepsinogen II monoclonal antibody, and a calibrator and a sample diluent containing a pepsinogen II antigen. A blocker component is contained in the sample diluent. The blocker is one or more human anti-mouse antibodies which neutralizes interfere of the HAMA. The method has the advantages that the high-sensitivity chemiluminescence technology and the magnetic particle preparation technology are combined, and an AutoLumo full-automatic detection analyzer is used in cooperation, so that a full-automatic detection is realized; meanwhile, the blocker component is added into the sample diluent, so that the heterophile antibody in the sample can be eliminated, and the anti-interference capability of the kit is greatly improved.

The invention discloses a pepsinogen I and pepsinogen II combined detection kit and application thereof. The invention discloses a pepsinogen I and II combined detection kit for the first time. The pepsinogen I and II combined detection kit comprises a coating antibody and a labeled antibody, wherein the coating antibody is an anti-PGI monoclonal antibody named as a monoclonal antibody IA and an anti-PGII monoclonal antibody named as a monoclonal antibody IIA; wherein the labeled antibody is an anti-PG I monoclonal antibody named as a monoclonal antibody IB and an anti-PG II monoclonal antibody named as a monoclonal antibody IIB. The invention further discloses application of the kit. According to the invention, the pepsinogen I and II combined detection kit is combined with a high-sensitivity inductively coupled plasma mass spectrometry detection technology on the basis of time-resolved fluorescence immunoassay. Detection of two indexes of PGI and PGII can be completed at the same time in one cycle, the time for clinically obtaining a PGI value and a PGI/PGII ratio result is shortened, and the detection cost is greatly reduced.

The invention relates to a pepsinogen II detection method and a kit, and especially relates to a dual wavelength fluorescence immunity chromatography detection method of pepsinogen II and a detection kit thereof. The method comprises the following steps: 1)preparing an immunity chromatography test strip; 2)preparing a freeze-dried probe; 3)preparing a sample weak solution; and 4)examining the sample. The pepsinogen II detection method and the kit have the advantages of high sensitivity, high accuracy, simple operation and low cost.

The invention discloses a pepsinogen II (PGII) quantitative assaying kit. The kit comprises a calibrated material, a quality control material, a resistance reagent, a magnetic particle reagent and a light emitting substrate. The invention further discloses a preparing method of the kit and a method for detecting PGII through the kit. According to the kit, the preparing method and the method, the resistance reagent is prepared from a PGII coated antibody labeled through fluorescein isothiocyanate and a PGII labeled antibody labeled through alkaline phosphatase, the magnetic particle reagent is prepared in the mode that the fluorescein isothiocyanate resistant antibody is coupled with carboxyl magnetic beads, even mixing and separation of an immunoreaction are easier accordingly, and the reaction speed is greatly increased; the novel chemical light emitting substrate APLS serves as the substrate, and therefore the sensitivity and the specificity of the kit are improved. The detecting kit is reliable in performance, high in sensitivity, wide in linear range and capable of being used in cooperation with semi-automatic and full-automatic instruments.

The invention relates to a pepsinogen I, pepsinogen II and gastrin-17 composite quality control product. The composite quality control material comprises a protein preserving solution, and pepsinogenI, pepsinogen II and gastrin-17 which are dissolved in the protein preserving solution, wherein the total volume of the protein preserving solution is calculated; wherein the protein preserving solution contains 0.8 to 3.5 g/L of AEP-HBC, 0.002 to 0.006 g/L of fibrinogen, 0.05 to 0.15 g/L of gelatin, 2 to 8 g/L of trehalose, 0.05 to 0.25 g/L of tetrapolyphosphate, 5-25 mmol/L of 2-(N-morpholine) ethanesulfonic acid, 0.5-4 g/L of ethylenediaminetetraacetic acid or ethylenediaminetetraacetic acid salt, 1-20 mmol/L of cysteine, methionine or arginine and 0.05-0.10% of a preservative,. The pepsinogen I, pepsinogen II and gastrin-17 composite quality control product is initiated at home, and can be stably stored for 14 months at the temperature of 2-8 DEG C. The invention further discloses a preparation method of the pepsinogen I, pepsinogen II and gastrin-17 composite quality control product. The invention also provides a detection kit containing the composite quality control product, andthe detection kit can be used for clinical detection of gastric functions.

The invention discloses a combined detection method for pepsinogen I and pepsinogen II. The method comprises the steps of immunochromatography test strip assembly, antibody coating, sample diluent preparation, sample detection and the like. The combined detection method for pepsinogen I and pepsinogen II utilizes a fluorescence immunochromatographic technique and adopts a double-antibody sandwichmethod, can detect the content of pepsinogen I/II in serum or plasma 10 minutes after sample adding, has the advantages of simplicity, convenience and rapidness, and avoids the damage of X-ray to human body and the inconvenience of gastroscope; and meanwhile, through the detection of samples with different concentrations, the sensitivity is higher, and the correlation is good.

The invention belongs to the technical field of bioengineering and particularly relates to soluble secretory expression of a fusion protein of a human pepsinogen II (PG II) and a maltose binding protein (MBP) in escherichia coli, provides a preparation method of the fusion protein, and also discloses application of a recombinant BMP-PG II fusion protein to preparation of monoclonal antibodies and calibration materials of pepsinogen II kits. Through the adoption of elements such as a Ptac promoter, an malE signal peptide and the maltose-binding protein, the soluble secretory expression of PG II from periphery to inside of escherichia coli is implemented; the immunogen activity of the recombinant human PG II protein from a prokaryotic expression system is greatly improved; and the preparation cost of the recombinant PG II is effectively reduced.

The invention discloses a pepsinogen I and pepsinogen II combined detection kit and a preparation method and detection method thereof. On the basis of enzyme immunoassay, a high sensitivity chemiluminescence detection technology is adopted, PG I and PG II can be simultaneously measured in one circulation test; compared with a method that measures PG I and PG II individually in sequence, the PG I/PG II detection time is shortened in clinic; furthermore, universal reagents are used during whole detection process, the detection cost is largely reduced; a Hamliton automatic detection and analysisinstrument is used, automation (from sample loading to result analysis) is realized; the interference of manual operation is avoided maximally; the time and error of operation such as sample loading,and the like are both reduced; the repeatability is strong, the detection becomes faster, and the results are more stable and reliable.

The invention relates to the technical field of biology, in particular to a pepsinogen I monoclonal antibody and application thereof, and discloses a group of pepsinogen I monoclonal antibodies which can be used in a matched mode. The pepsinogen I monoclonal antibodies have high specificity and stability when being combined with PGI, cannot generate cross reaction with pepsinogen II, and are suitable for various methodologies, and high-sensitivity and high-precision detection of PGI is realized.

The invention relates to the technical field of biology, in particular relates to a pepsinogen II monoclonal antibody and application thereof, and discloses a group of pepsinogen II monoclonal antibodies which can be used in cooperation, which can be applied to different methodologies, accurate measurement of PGII is achieved, interference of other impurities can be avoided, and high-specificity binding is achieved. The monoclonal antibody provided by the invention can assist in research and development of medical diagnostic reagents, and has great significance.

The invention provides a stable protein solution. The protein solution comprises a buffer solution and a protein containing a heme auxiliary group or a protein conjugate of the protein containing theheme auxiliary group and other proteins. Ferrous ions in the heme auxiliary group are combined with carbon monoxide, so that the protein or the protein conjugate can be prevented from being oxidized and denatured in the storage process, and the stability of the protein solution is improved. The protein solution may also include other stabilizers. The protein solution disclosed by the invention canbe stably stored for at least one year at 2-8 DEG C, has good stability, and can be used for preparing a chemiluminescence detection kit or an enzyme-linked immunosorbent assay kit, such as a detection kit for detecting gastrin 17, pepsinogen I and pepsinogen II.

The invention discloses a chemiluminiscence detection kit for diagnosing gastropathy in a combined manner. The detection kit is mainly used for performing pepsinogen I detection and pepsinogen II detection. On the basis of the detection kit, the invention further provides a marker combined standard value, which is applicable to the detection kit and can provide reference for gastropathy detection, for detection. The detection kit has the advantages that the detection kit is wide in linearity range, high in specificity, convenient to operate, and the like in terms of gastropathy diagnosis and early gastric cancer screening; in addition, the detection kit is accurate and detailed in gastropathy diagnosis and high in market popularization value, and damage, caused by X-rays, on human bodies and the inconvenience of a gastroscope are avoided.

The invention discloses a CHO-K1 cell strain, which contains a gene capable of efficiently secreting and expressing PG II recombinant protein. The invention also discloses a method for preparing the CHO-K1 cell strain, and the method comprises the following steps: 1) cloning a gene sequence as shown in SEQ ID NO.1 into an eukaryotic expression vector to obtain a recombinant plasmid containing the gene encoding the PG II recombinant protein; (2) transfecting the recombinant plasmid into a CHO-K1 cell, so as to obtain a CHO-K1 cell strain; and 3) culturing, screening and domesticating the CHO-K1 cell strain in the step 2) to obtain the cell strain capable of efficiently secreting and expressing the PG II recombinant protein. The invention also discloses an anti-PG II monoclonal antibody, wherein the antigen epitope combined with a monoclonal antibody 1 is located at aa78-aa90 of the pepsinogen II; and the antigen epitope combined with a monoclonal antibody 2 is located at aa280-aa291 of the pepsinogen II. The protein provided by the invention is high in expression quantity, good in quality and low in cost; the monoclonal antibody can be paired and used for detecting the PG II protein, and is good in specificity and high in sensitivity.

The invention discloses a pepsinogen immunochromatographic detection kit based on carbon quantum dots, and a preparation method thereof. The pepsinogen immunochromatographic detection kit comprises aplastic plate and an adsorption layer fixed on the plastic plate, wherein the adsorption layer is composed of a sample pad, a glass cellulose film, a nitrocellulose film and a water absorbent pad which are in lap joint with each other from the test end in sequence; the glass cellulose film is coated with carbon quantum dot-labeled pepsinogen I and pepsinogen II; the nitrocellulose film is providedwith a detection line 1, a detection line 2 and a control line, the detection line 1 is coated with pepsinogen I secondary antibodies, the detection line 2 is coated with pepsinogen II secondary antibodies, and the control line is coated with goat-anti-mouse IgG; and the carbon quantum dots are respectively modified by means of ethylenediamine and N-hydroxysuccinimide. The pepsinogen immunochromatographic detection kit of the invention can be used for combined detection of the pepsinogen I and the pepsinogen II, and has the advantages of detecting pepsinogen in serum efficiently in a low-toxicity manner.

The invention provides a pepsinogen II (PGII) detection kit. The pepsinogen I detection kit comprises a reagent R1 and a reagent R2 which are independent to each other, wherein the reagent R1 comprises the following components: 1 to 10g/L of a trihytdroxy methyl aminomethane buffering solution, 1 to 20g/L of sodium chloride, 0.1 to 10mg/L of Tween 20, 10 to 50g/L of polyethylene glycol 20000 and 0.1 to 10g/L of sodium azide; the reagent R2 comprises the following components: 1 to 10g/L of the trihytdroxy methyl aminomethane buffering solution, 10 to 100g/L of trehalose, 2 to 10ml/L of a surfactant, 10 to 1000mg/L of a pepsinogen II (PGII)antibody and 0.1 to 10g/L of the sodium azide. The pepsinogen II (PGII) detection kit is ready to use and does not need to be prepared; and the pepsinogenII (PGII) detection kit has high detection sensitivity and good measurement repeatability and has relatively good stability when being stored for a long period. The invention further provides a production technology of the pepsinogen II (PGII) detection kit.

The invention relates to a stomach function and stomach cancer risk detection device and a preparation method thereof, and belongs to the field of medical detection equipment. The device is prepared by adhering a nitrocellulose membrane with a solid phase containing high-specificity gastrin 17, pepsinogen I, pepsinogen II antibody, anti-human IgG antibody, anti-human IgM antibody and goat-anti-mouse IgG polyclonal antibody, a glass fiber membrane adsorbed with colloidal gold labeled G-17, PGI and PGII antibodies and HP urease antigens, a sample pad, absorbent paper and other auxiliary materials. According to the invention, on the basis of ensuring complete release of immune colloidal gold, the reaction sensitivity is effectively improved; under the same threshold value, the dosage of immune colloidal gold can be reduced, the cost is saved, five gastric function evaluation and gastric cancer risk markers of G-17, PGI, PGII, HP urease IgG antibodies and IgM antibodies in a specimen can be detected at the same time, and the complexity of production operation is not increased. The test paper is high in sensitivity, strong in specificity, simple and convenient to operate, time-saving and strong in practicability.