CN103713140A - Latex immunoturbidimetry type pepsinogen II detection kit capable of eliminating chyle interference - Google Patents
Latex immunoturbidimetry type pepsinogen II detection kit capable of eliminating chyle interference Download PDFInfo
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- CN103713140A CN103713140A CN201410010900.4A CN201410010900A CN103713140A CN 103713140 A CN103713140 A CN 103713140A CN 201410010900 A CN201410010900 A CN 201410010900A CN 103713140 A CN103713140 A CN 103713140A
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Images
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/914—Hydrolases (3)
- G01N2333/948—Hydrolases (3) acting on peptide bonds (3.4)
- G01N2333/95—Proteinases, i.e. endopeptidases (3.4.21-3.4.99)
- G01N2333/964—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue
- G01N2333/96425—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals
- G01N2333/96427—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals in general
- G01N2333/9643—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals in general with EC number
- G01N2333/96472—Aspartic endopeptidases (3.4.23)
- G01N2333/96475—Aspartic endopeptidases (3.4.23) with definite EC number
- G01N2333/96477—Pepsin (3.4.23.1; 3.4.23.2; 3.4.23.3)
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- Urology & Nephrology (AREA)
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- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
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- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
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- Investigating Or Analysing Biological Materials (AREA)
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Abstract
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.
Description
Technical field
The invention belongs to medical test technical field, relate to a kind of Serum Pepsinogen II detection kit of eliminating the latex immunoturbidimetry of chyle interference.
Background technology
Propepsin (Pepsinogen, PG) is pepsic inactive precursor in gastric juice, is a kind of L-aminobutanedioic acid proteinase precursor of gastric secretion, is the single chain polypeptide of molecular weight 42000Da, changes the activated pepsin of tool in stomach into.
According to distributing in biochemical property, immunogenicity, cell derived and tissue, propepsin can be divided into propepsin I (PG I), two subgroups of PGⅡ (PG II): wherein 1～5 component immunogenicity is approximate, be called PG I (PGA), mainly chief cell and the mucous neck cells by fundus gland secretes; 6～7 components are called as PG II (PGC), and except by above-mentioned two kinds of emiocytosises, the Brunner gland of the mucilage cell of pyloric gland, cardiac gland and duodenum epimere also can produce PG II, and the synthetic PG II of gastric mucosa is about 25% of total amount.
Under normal circumstances, the propepsin major part that gastric mucosa produces enters gastral cavity, meets the rear peptide chain of acid and decomposes activation for pepsin, brings into play the effect of its digesting protein; Fraction pepsin principle sees through gastric mucosa capillary and enters blood circulation.Serum Pepsinogen concentration can reflect the secretion level of gastric mucosa, and different propepsin concentration levels also can reflect that the function and morphology of different parts gastric mucosa changes.
Japanese health care for the aged method has been listed in the detection of propepsin in, applies this project and carries out large-area mass survey and make the rate of examining morning of Japanese cancer of the stomach bring up to 90%, is called as " the serology biopsy " of fundus gland mucous membrane in Japan and Korea S.
The method that detects at present Serum Pepsinogen content mainly contains enzyme linked immunosorbent assay (ELISA) (ELISA), radiommunoassay (RIA), chemoluminescence method and latex immunoturbidimetry etc.
ELISA method accurately, quantitatively, have very high susceptibility, but running program is loaded down with trivial details, and length consuming time can not adapt to the needs of extensive health check-up examination or emergency treatment; RIA method is highly sensitive, high specificity, but radio-immunity reaction is emulative reaction, and the value recording is relative quantity but not absolute magnitude, and has radiation and radioactive contamination; Chemoluminescence method degree of accuracy is high, and detection speed is fast, but with high costs, auxiliary facility is required high, cannot be applied to vast in, hospital and the medical and health organization of basic unit; Latex immunoturbidimetry has change fast and automatically easy and simple to handle, quantitatively accurate, susceptibility advantages of higher, can meet the needs such as Outpatient and emergency and extensive health check-up examination.
Existing latex immunoturbidimetry reagent, because it detects the turbidity that principle is the immune complex of assaying reaction formation, is therefore very easily subject to the impact that chyle disturbs.The method of this problem of reply, is mainly to use special chyle remover to carry out pre-service to sample at present.And when large-scale mass survey, health examination examination and clinical patient check; often can run into the muddy samples such as a large amount of chyles, haemolysis; now by purchasing sample process agent; these samples are processed separately; will greatly increase operating personnel's workload; improve operation cost, also for the organization arrangement of clinical examination brings a difficult problem.
Therefore, develop a kind of PGⅡ detection kit that can eliminate the latex immunoturbidimetry of chyle interference, seem very necessary.
Summary of the invention
The present invention is in order to overcome above-mentioned prior art defect, the kit of PGⅡ content in a kind of human body serum is provided, do not need again chyle sample to be processed separately, saved manpower, resource, used simple, quick, highly sensitive, the demand that not only can adapt to large-scale crowd generaI investigation, also can meet daily clinical fast, the requirement that checks of high flux, be applicable to industrialization, be convenient to clinical expansion.The technical scheme that the present invention takes is a kind of latex immunoturbidimetry PGⅡ detection kit, and its composition comprises:
Reagent 1, be a kind of for kit provides appropriate reaction environment, eliminates chyle and disturb, make antigen to keep native conformation, and control reaction and reach the time of terminal and the reagent of speed, comprise electrolyte, chyle remover, set accelerator, stabilizing agent, surfactant, antiseptic and damping fluid;
Reagent 2, is a kind of milky white liquid of the homogeneous that contains the coated latex particle of anti-human PGⅡ antibody, comprises anti-human PGⅡ antibody coated latex particle, antiseptic and damping fluid;
Calibration object, is the human serum matrix damping fluid that adds variable concentrations PGⅡ antigen, is used for relatively carrying out result calculating with sample, comprises electrolyte, antiseptic, stabilizing agent, PGⅡ antigen, human serum and damping fluid.
Described electrolyte is selected from inorganic salts, preferably sodium chloride, potassium chloride, magnesium chloride or magnesium sulfate.
Described chyle remover, is selected from polyglycol-sulfuric acid chrysanthemum glycan azoviolet of lipase or 1～5%.
Described set accelerator is selected from PEG-4000, PEG-6000, PEG-8000, sodium dextran sulfate.
Described stabilizing agent is selected from bovine serum albumin(BSA) (BSA), casein, fishskin gelatin albumen (Fish skingelatin), sorbierite, disodium ethylene diamine tetraacetate (EDTA).
Described surfactant is selected from tween series, fatty alcohol polyglycol ether, polyoxyethylene phenyl ether, polyoxyethylene laurel ether series.
Described antiseptic is selected from Sodium azide, phenol, P-hydroxybenzoic acid, ethyl-para-hydroxybenzoate, Proclin series.
Described damping fluid is selected from Tris-HCl damping fluid, glycocoll-NaOH damping fluid, phosphate buffer, HEPES-NaOH damping fluid, MOPSO damping fluid.
Described antibody is monoclonal antibody and the mixing of polyclonal antibody, and the specificity of monoclonal antibody reactive is stronger, but sensitvity constraint is added a small amount of polyclonal antibody and can be improved the sensitivity of reaction.Antibody type is selected from IgG antibody or IgY antibody, and antibody sources is selected from that rabbit is anti-human, goat is anti-human, sheep is anti-human, mouse-anti people, chicken is anti-human, duck is anti-human.
Described latex particle is polystyrene latex, and diameter range is 150～250nm.The finishing of latex particle has functional group, by this functional group, combines with antibody again, can alleviate the obstruction that steric configuration forms covalent bond, and antibody is more easily linked on latex particle, and the activity of antibody is higher, can significantly improve sensitivity for analysis.The functional group of modifying can be selected a kind of of carboxyl, amino, hydroxyl, hydrazides, chloromethyl.
The present invention's coated antibody used is chemical crosslink technique to the method on latex particle surface, the method is carried out in crosslinked damping fluid by chemical cross-linking agent, described chemical cross-linking agent is selected a kind of in carbonization imines (EDAC), N-hydroxy-succinamide (NHS), N-hydroxy thiosuccinimide (Sulfo-NHS), hydrazides, isocyanates, described crosslinked damping fluid is selected from different amino damping fluids, MES, MOPSO, MOPS, HEPES damping fluid, pH is between 6.0～8.0.
Damping fluid in described PGⅡ detection kit reagent 1, requires its surge capability for regulating pH between 7.0～9.0, can select above-mentioned damping fluid wherein a kind of, and concentration range is 10～100mmol/L.Chyle remover requires to have stronger chyle to remove ability and certain reactionlessness, can select above-mentioned chyle remover wherein a kind of, preferred fat enzyme, concentration range 5～50KU/L.Electrolyte is selected from inorganic salts, can maintain solution osmotic pressure, is conducive to keep the stability of Proteins In Aqueous Solutions, can select above-mentioned electrolyte wherein a kind of, preferred sodium chloride, and concentration range is 1%～5%.Set accelerator can be accelerated immune response speed, shortens detection time, can select above-mentioned set accelerator wherein a kind of, preferred PEG-6000, and concentration range is 2%～6%.Stabilizing agent can provide well stablizes reaction environment, makes antigen keep native conformation, can select aforementioned stable agent wherein a kind of, preferred BSA, and concentration range is 0.5%～5%.Surfactant can be when reagent 2 joins reaction system, be adsorbed onto rapidly the surface of latex particle, reduce latex particle surface tension, make latex particle be difficult for assembling, effectively improve dispersiveness and the stability of latex particle in solution, can select above-mentioned surfactant wherein a kind of, preferred polysorbas20, concentration range is 0.1%～1.0%.Select to add foregoing preservatives wherein a kind of, concentration range is 0.05%～0.1% again.
Damping fluid in described PGⅡ detection kit reagent 2, requires its surge capability for regulating pH between 7.0～9.0, can select above-mentioned damping fluid wherein a kind of, and concentration range is 10～100mmol/L.Anti-human PGⅡ antibody, wherein the ratio range of monoclonal antibody/how anti-is 5～10, and antibody type is selected from IgG, IgY antibody, and antibody sources can select above-mentioned antibody wherein a kind of, preferably the anti-human IgY antibody of duck.Latex particle finishing has functional group, can select the functional group of above-mentioned modification wherein a kind of, preferably modifies carboxyl.Chemical cross-linking agent selects above-mentioned chemical cross-linking agent wherein a kind of, and preferably EDAC, is cross-linked damping fluid and selects above-mentioned crosslinked damping fluid wherein a kind of, preferred HEPES damping fluid, and the antibody latex granule density scope of its sensitization is 0.5～5mg/ml.Select to add foregoing preservatives wherein a kind of, concentration range is 0.05%～0.1% again.
Described PGⅡ detection kit calibration object, comprise the PG II of content between 0～100ng/ml, wherein PG II calibration object level 1 is that PG II calibration object level 2,3,4 is for adding the human serum matrix damping fluid of different PG II content not containing the human serum matrix damping fluid of PG II.Damping fluid can select above-mentioned damping fluid wherein a kind of, and concentration range is 10～100mmol/L, and pH is 6.5～8.5, and human serum ratio is 5%.The PG II of adding select to derive from from human body fluid extraction, purifying and natural PG II, or from the cultivation of duodenum JEG-3, purifying and PG II, its purity >=95%; The electrolyte adding selects above-mentioned electrolyte wherein a kind of, preferred sodium chloride, and concentration range is 1%～5%; The stabilizing agent adding selects aforementioned stable agent wherein a kind of, and concentration range is 0.5～5%; Select to add foregoing preservatives wherein a kind of, concentration range is 0.01%～0.1% again.
Percent concentration described in the present invention, as do not carry out specified otherwise, be quality concentration of volume percent, i.e. the grams of contained solute in 100ml solution.
Kit of the present invention, adopts latex enhancing immune turbidimetry quantitatively to detect PGⅡ content in human serum.Its principle is: the PGⅡ in tested sample be coated on anti-human PGⅡ monoclonal and the polyclonal antibody generation antigen-antibody reaction on latex particle, the compound of the Ag-Ab-latex particle forming is woven into netted and aggegation occurs, cause turbidity to rise, while keeping in reaction system antibody excess, detect this turbidity in the absorbance at 700nm wavelength place, contrast calibration curve can be obtained the content of PGⅡ in sample.
Compared with prior art, tool of the present invention has the following advantages and effect: 1, anti-chyle interference performance is strong, does not need sample to carry out pre-service, has saved manpower, resource, can be applied to large-scale health check-up examination and mass survey, be conducive to extensively carrying out of project; 2, the sensitivity of kit of the present invention is higher, and the measured value that detects low concentration sample is more accurate; 3, kit of the present invention and euzymelinked immunosorbent assay (ELISA) (ELISA) contrast the data analysis detecting to clinical sample, and correlativity is good, and clinical meaning is remarkable; 4, detection means is simple, less demanding to medical facilities, can be applied to various full-automatic, semi-automatic biochemical analyzers.
Accompanying drawing explanation
Fig. 1 is for adopting 4 different gradients of the embodiment of the present invention 1 preparation, and concentration is respectively 0,20,40, the PGⅡ calibration object of 80ng/ml, on Hitachi's 7080 automatic clinical chemistry analyzers, draws out the typical curve of PG II calibration object of the present invention.Wherein X-axis represents the content (ng/ml) of PGⅡ; Y-axis represents absorbance.
Embodiment
Embodiment 1
One, eliminate the preparation of the PGⅡ detection kit of chyle interference
The preparation of reagent 1: in glycocoll-NaOH of 50mmol/L, pH7.5 damping fluid, add lipase, 1.2% sodium chloride, 0.3% polysorbas20,5%PEG-6000,0.5%BSA, the 0.05%Proclin300 of 20KU/L, stir and regulate pH to 8.0, the PGⅡ detection kit reagent 1 that the chyle that is eliminated disturbs.
The preparation of reagent 2:
(1) antibody linked: after the HEPES-NaOH damping fluid dilution of the anti-human PGⅡ IgY of the mixing duck antibody that is 5:1 by 0.5mg monoclonal antibody/how anti-ratio with 5ml20mmol/L, the polystyrene latex solution that adds finishing carboxyl, diameter 150nm, add again 1.5mg EDAC, under 37 ℃ of conditions, react 8h, add the Tris-HCl damping fluid of 0.5ml100mmol/L to stir cessation reaction after 1h;
(2) latex cleans: centrifugal removal supernatant to be to remove the accessory substance etc. of unnecessary antibody, crosslinking chemical and cross-linking reaction,
Bottom settlings is the latex being coated with.With glycocoll-NaOH damping fluid of 20ml50mmol/L, pH8.0, wash 3 times;
(3) latex suspends: glycocoll-NaOH damping fluid that 20ml is same joins above-mentioned precipitation, ultrasound wave is processed 5min, mix to obtain the milky latex suspension of homogeneous, the antibody latex granule density that makes sensitization is 2mg/ml, add again 0.05% Proclin300, the PGⅡ detection kit reagent 2 that the chyle that is eliminated disturbs.
The preparation of PGⅡ calibration object: pH6.8, contain 20mmol/L Tris-HCl cushion pre-service human serum matrix damping fluid in, human serum ratio is 5%, add that concentration is respectively 0,20,40,80ng/ml from human body fluid extraction, purifying and, the PGⅡ antigen of purity >=95%, stir, as PGⅡ multiple spot calibration object.In addition, in pretreated human serum matrix damping fluid, also comprise 2.5% sodium chloride, 0.8%EDTA, 0.02%Proclin300.
Two, eliminate the method for the PGⅡ detection kit detection sample of chyle interference
Detection method and experiment parameter are as follows:
Analytical approach: Two point end assay;
The consumption of reagent: reagent 1 and reagent 2 consumptions are respectively 160 μ l and 30 μ l;
The consumption of sample: 6 μ l;
Detect wavelength: 700nm.
Detecting step: 160 μ l reagent 1 add 6 μ l samples adds the rear beginning read point of 30 μ l reagent 2 after 37 ℃ of 5min, after reaction 5min, reads another point, obtains the difference of absorbance.
The typical curve of PGⅡ detection kit calibration object prepared by drafting the present embodiment:
4 kinds of different contents that adopt the present embodiment to prepare, be respectively 0,20,40, the PGⅡ calibration object of 80ng/ml, on Hitachi's 7080 automatic clinical chemistry analyzers, according to above-mentioned detecting step, record the typical curve (as shown in Figure 1) of PG II calibration object of the present invention.Each point in Fig. 1 on curve represents the calibration object of a content.Wherein X-axis represents the content (ng/ml) of PGⅡ; Y-axis represents absorbance.
Get sample to be tested, according to above-mentioned detecting step, detect equally the absorbance difference of sample, substitution calibration curve, can calculate the content of PGⅡ in sample to be tested.If the concentration of PGⅡ exceeds calibration curve scope in sample, the accuracy in order to guarantee to detect, detects after need to suitably diluting sample again.
Three, eliminate the anti-chyle interference performance of the PGⅡ detection kit of chyle interference
According to following method of operating, detect the anti-chyle jamming performance of PGⅡ detection kit:
(1) collect the close human serum sample without haemolysis, jaundice, chyle, turbid phenomenon of measured value of lower scope, mix, this PHS's sample is measured, the value that obtains its PGⅡ is 8.6ng/ml;
(2) according to shown in following table 1, table 2, in PHS, add the chyle chaff interference of variable concentrations gradient;
(3) use respectively certain producer's immunoturbidimetry PGⅡ detection kit (in this test, be called for short kit A) and the PG II detection kit prepared of the present embodiment (in this test, be called for short kit B), to having added the serum sample of different chyle units concentration, carry out blank determination, repeat 3 times, computation of mean values, CV and deviation, deviation range is considered as noiselessly ± 10% with interior, and deviation range surpasses ± 10% and is considered as disturbing.
Table 1 kit A detects the result of adding variable concentrations chyle chaff interference serum sample
Table 2 kit B detects the result of adding variable concentrations chyle chaff interference serum sample
The demonstration of table 1 result, with the range of chyle turbidity from 0 to 8000, the detected value deviation of kit A also increases thereupon, and when chyle turbidity unit is greater than 3000, when detected value and interpolation interference, the deviation of detected value is all greater than 10%, is judged to be existence and disturbs.
The demonstration of table 2 result, with the range of chyle turbidity from 0 to 8000, the variation of the detected value deviation of kit B is less, and when detected value and interpolation interference, the deviation of detected value, all in ± 10%, is judged to be and does not have interference.
This test has shown compares existing immunoturbidimetry method kit, and PGⅡ detection kit of the present invention has obviously improved the antijamming capability to chyle.
Four, eliminate the sensitivity of the PGⅡ detection kit of chyle interference
According to following method of operating, detect the sensitivity of PGⅡ detection kit:
(1) determine that a reagent can detect low concentration value accurately and compare with water is blank, the sample of 1.7ng/ml is selected in this test;
(2) detect 20 water, record absorbance numerical value, calculating mean value (X water) and standard deviation (SD);
(3) sample that 20 concentration of detection are 1.7ng/ml, records absorbance numerical value, calculating mean value (X1.7) and standard deviation (SD);
(4) the absorbance mean value of water adds that 3SD is as absorbance corresponding to lowest detectable limit, because the relation of absorbance and concentration is linear relationship substantially, the concentration that can relatively calculate by the absorbance mean value with 1.7ng/ml sample lowest detectable limit is sensitivity.Formula is 1.7 * (X water+3SD)/X1.7.
The data result of the sensitivity test of the PGⅡ detection kit of preparing in the present embodiment (in this test, being called for short " kit of the present invention ") and contrast agents box (certain producer's immunoturbidimetry PGⅡ detection kit) is as shown in table 3.
In following table, △ A represents the difference of absorbance.
Table 3 kit of the present invention and contrast agents box Sensitivity comparison data
Sensitivity=1.7 of kit of the present invention * (22+3 * 1.4)/698.8=0.06ng/ml.
Sensitivity=1.7 of contrast agents box * (24.5+3 * 11.9)/561.5=0.18ng/ml.
Result shows that the sensitivity of kit of the present invention can reach 0.06ng/ml, is better than contrast agents box.
Kit of the present invention is by being attached to a certain proportion of PG II monoclonal antibody and polyclonal antibody on the latex particle of carboxyl modified with chemical crosslink technique, make on the one hand antigen-antibody binding reaction more rapidly, firmly, by latex agglutination effect, amplify on the other hand the detection progression of reaction, thereby realized the sensitivity raising of kit.From the result of above-mentioned table 3, kit of the present invention highly sensitive in the sensitivity of contrast agents box, visible kit of the present invention has been realized the effect that sensitivity improves.
Five, eliminate the PGⅡ detection kit of chyle interference and the correlativity that enzyme is exempted from method detection kit
PG II detection kit prepared by use the present embodiment is (in this test, be called for short " PG II kit of the present invention ") and the latex immunoturbidimetry PG of our company I detection kit (in this test, be called for short " PG of our company I kit "), adopt Hitachi's 7080 automatic clinical chemistry analyzers, 50 parts of Freshman serum without chyle, haemolysis, jaundice, turbid phenomenon are detected, carry out correlation analysis with the testing result of contrast agents box (the propepsin I of Beijing producer euzymelinked immunosorbent assay (ELISA), II detection kit are called for short " enzyme is exempted from kit ").PG II kit of the present invention detects according to above-mentioned " two, eliminate the method that PGⅡ detection kit that chyle disturbs detects sample ", the PG of our company I kit detects to specifications, enzyme is exempted from kit and is detected according to its instructions, and measurement result is as shown in table 4.
In following table, the term of reference that enzyme is exempted from method kit is: PG I (70-200ng/ml); PG II (0-15ng/ml); PG I/PG II >7.5; The term of reference of our company's latex immunoturbidimetry method kit is: PG I (70-240ng/ml); PG II (0-20ng/ml); PG I/PG II >3.
Table 4 kit of the present invention and enzyme are exempted from the clinical data of comparing of method kit
By upper table statistics, can be found out, in this 50 routine fresh patients serum, there are altogether 35 routine negative findingses and 15 routine positive findingses, it is one to one that enzyme is exempted from method kit and kit of the present invention in result judgement, and two individual event results and ratio thereof all exist coincideing in clinical tolerance band.Although two kinds of square ratio juris and term of reference have certain difference, judge from the angle of clinical practice, the correlativity of the two is good, and clinical meaning is remarkable.
In addition, above test is that the 7080 model automatic clinical chemistry analyzers that adopt Hitachi, Ltd to manufacture carry out, but the application of reagent of the present invention is not limited to above-mentioned instrument, also be applicable to other full-automatic or semi-automatic biochemical analyzers, and those skilled in the art can according to circumstances appropriately adjust to location parameter.
Embodiment 2
One, eliminate the preparation of the PGⅡ detection kit of chyle interference
The preparation of reagent 1: in the Tris-HCl of 20mmol/L, pH7.5 damping fluid, add lipase, 4.5% potassium chloride, 0.6% polyoxyethylene phenyl ether, 2%PEG-6000,2.5%EDTA, 0.08% Sodium azide of 5KU/L, stir and regulate pH to 7.2, the PGⅡ detection kit reagent 1 that the chyle that is eliminated disturbs.
The preparation of reagent 2:
(1) antibody linked: after the MOPSO-HCl damping fluid dilution of the anti-human PGⅡ IgG of the mixing rabbit antibody that is 6.5:1 by 0.5mg monoclonal antibody/how anti-ratio with 5ml10mmol/L, the polystyrene latex solution that adds finishing amino, diameter 200nm, add again 2.0mg NHS, under 37 ℃ of conditions, react 8h, add glycocoll-NaOH damping fluid of 0.5ml80mmol/L to stir cessation reaction after 1h;
(2) latex cleans: centrifugal removal supernatant is to remove the accessory substance etc. of unnecessary antibody, crosslinking chemical and cross-linking reaction, and bottom settlings is the latex being coated with.With the Tris-HCl damping fluid of 20ml20mmol/L, pH7.2, wash 3 times;
(3) latex suspends: the Tris-HCl damping fluid that 80ml is same joins above-mentioned precipitation, ultrasound wave is processed 5min, mix to obtain the milky latex suspension of homogeneous, the antibody latex granule density that makes sensitization is 0.5mg/ml, add again 0.08% Sodium azide, the PGⅡ detection kit reagent 2 that the chyle that is eliminated disturbs.
The preparation of calibration object: pH7.8, contain 100mmol/L HEPES-NaOH cushion pre-service human serum matrix damping fluid in, human serum ratio is 5%, add that concentration is respectively 0,20,40,80ng/ml from human body fluid extraction, purifying and, the PGⅡ antigen of purity >=95%, stir, as PGⅡ multiple spot calibration object.In addition, in pretreated human serum matrix damping fluid, also comprise 5% potassium chloride, 2%BSA, 0.05% Sodium azide.
The PGⅡ detection kit of preparing in the present embodiment detects the method for sample, with " eliminating the method for the PGⅡ detection kit detection sample of chyle interference " in embodiment 1.
Two, eliminate the anti-chyle interference performance of the PGⅡ detection kit of chyle interference
According to following method of operating, detect the anti-chyle jamming performance of PGⅡ detection kit:
(1) collect the close human serum sample without haemolysis, jaundice, chyle, turbid phenomenon of measured value of lower scope, mix, this PHS's sample is detected, the value that obtains its PGⅡ is 8.6ng/ml;
(2) according to shown in following table 5, table 6, in PHS, add the chyle chaff interference of variable concentrations gradient;
(3) use respectively certain producer's immunoturbidimetry PGⅡ detection kit (in this test, be called for short kit A) and the PG II detection kit prepared of the present embodiment (in this test, be called for short kit B), to having added the serum sample of different chyle units concentration, carry out blank determination, repeat 3 times, computation of mean values, CV and deviation, deviation range is considered as noiselessly ± 10% with interior, and deviation range surpasses ± 10% and is considered as disturbing.
Table 5 kit A detects the result of adding variable concentrations chyle chaff interference serum sample
Table 6 kit B detects the result of adding variable concentrations chyle chaff interference serum sample
The demonstration of table 5 result, with the range of chyle turbidity from 0 to 8000, the detected value deviation of kit A also increases thereupon, and when chyle turbidity unit is greater than 3000, when detected value and interpolation interference, the deviation of detected value is all greater than 10%, is judged to be existence and disturbs.
The demonstration of table 6 result, with the range of chyle turbidity from 0 to 8000, the variation of the detected value deviation of kit B is less, and when detected value and interpolation interference, the deviation of detected value, all in ± 10%, is judged to be and does not have interference.
This test has shown compares existing immunoturbidimetry method kit, and PGⅡ detection kit of the present invention has obviously improved the antijamming capability to chyle.
Three, eliminate the sensitivity of the PGⅡ detection kit of chyle interference
According to following method of operating, detect the sensitivity of PGⅡ detection kit:
(1) determine that a reagent can detect low concentration value accurately and compare with water is blank, the sample of 1.7ng/ml is selected in this test;
(2) detect 20 water, record absorbance numerical value, calculating mean value (X water) and standard deviation (SD);
(3) sample that 20 concentration of detection are 1.7ng/ml, records absorbance numerical value, calculating mean value (X1.7) and standard deviation (SD);
(4) the absorbance mean value of water adds that 3SD is as absorbance corresponding to lowest detectable limit, because the relation of absorbance and concentration is linear relationship substantially, the concentration that can relatively calculate by the absorbance mean value with 1.7ng/ml sample lowest detectable limit is sensitivity.Formula is 1.7 * (X water+3SD)/X1.7.
The data result of the sensitivity test of the PGⅡ detection kit of preparing in the present embodiment (in this test, being called for short " kit of the present invention ") and contrast agents box (certain producer's immunoturbidimetry PGⅡ detection kit) is as shown in table 7.
In following table, △ A represents the difference of absorbance.
Table 7 kit of the present invention and contrast agents box remolding sensitivity are to data
Sensitivity=1.7 of kit of the present invention * (22.3+3 * 1.9)/699.3=0.07ng/ml.
Sensitivity=1.7 of contrast agents box * (24.55+3 * 12.0)/564.35=0.18ng/ml.
Result shows that the sensitivity of kit of the present invention can reach 0.07ng/ml, is better than contrast agents box.
Kit of the present invention is by being attached to a certain proportion of PG II monoclonal antibody and polyclonal antibody on the latex particle of carboxyl modified with chemical crosslink technique, make on the one hand antigen-antibody binding reaction more rapidly, firmly, by latex agglutination effect, amplify on the other hand the detection progression of reaction, thereby realized the sensitivity raising of kit.From the result of above-mentioned table 7, kit of the present invention highly sensitive in the sensitivity of contrast agents box, visible kit of the present invention has been realized the effect that sensitivity improves.
Four, eliminate the PGⅡ detection kit of chyle interference and the correlativity that enzyme is exempted from method detection kit
PG II detection kit prepared by use the present embodiment is (in this test, be called for short " PG II kit of the present invention ") and the latex immunoturbidimetry PG of our company I detection kit (in this test, be called for short " PG of our company I kit "), adopt Hitachi's 7080 automatic clinical chemistry analyzers, 50 parts of Freshman serum without chyle, haemolysis, jaundice, turbid phenomenon are detected, carry out correlation analysis with the testing result of contrast agents box (the euzymelinked immunosorbent assay (ELISA) propepsin I of Beijing producer, II detection kit are called for short " enzyme is exempted from kit ").PG II kit of the present invention detects according to above-mentioned " PGⅡ detection kit detects the method for sample ", and the PG of our company I kit detects to specifications, and enzyme is exempted from kit and detected according to its instructions, and testing result is as shown in table 8.
In following table, the term of reference that enzyme is exempted from method kit is: PG I (70-200ng/ml); PG II (0-15ng/ml); PG I/PG II >7.5; The term of reference of our company's latex immunoturbidimetry method kit is: PG I (70-240ng/ml); PG II (0-20ng/ml); PG I/PG II >3.
Table 8 reagent of the present invention and enzyme are exempted from method reagent clinical comparison data
By upper table statistics, can be found out, in this 50 routine fresh patients serum, occur altogether 30 routine negative findingses and 20 routine positive findingses, be excused from an examination agent and reagent of the present invention of enzyme is one to one in result judgement, and two individual event results and ratio thereof all exist coincideing in clinical tolerance band.Although two kinds of square ratio juris and term of reference have certain difference, judge from the angle of clinical practice, the correlativity of the two is good, and clinical meaning is remarkable.
Embodiment 3
One, eliminate the preparation of the PGⅡ detection kit of chyle interference
The preparation of reagent 1: in the phosphate buffer of 80mmol/L, pH8.5, add 4% polyglycol-sulfuric acid chrysanthemum glycan magnesium, 2.5% magnesium sulfate, 1% Tween 80,6%PEG-8000,5% sorbierite, 0.1%Proclin300, the PGⅡ detection kit reagent 1 that the chyle that is eliminated disturbs.
The preparation of reagent 2:
(1) antibody linked: after the phosphate buffer dilution of the anti-human PGⅡ IgY of the mixing chicken antibody that is 9:1 by 0.5mg monoclonal antibody/how anti-ratio with 5ml50mmol/L, the polystyrene latex solution that adds finishing chloromethyl, diameter 250nm, add again 1.5mg EDAC, under 37 ℃ of conditions, react 8h, add the Tris-HCl damping fluid of 0.5ml80mmol/L to stir cessation reaction after 1h;
(2) latex cleans: centrifugal removal supernatant is to remove the accessory substance etc. of unnecessary antibody, crosslinking chemical and cross-linking reaction, and bottom settlings is the latex being coated with.With the phosphate buffer of 20ml, 80mmol/L, wash 3 times;
(3) latex suspends: the Tris-HCl damping fluid that 10ml is same joins above-mentioned precipitation, ultrasound wave is processed 5min, mix to obtain the milky latex suspension of homogeneous, the antibody latex granule density that makes sensitization is 4mg/ml, add again 0.1% Proclin300, the PGⅡ detection kit reagent 2 that the chyle that is eliminated disturbs.
The preparation of calibration object: pH8.4, contain 50mmol/L phosphate-buffered thing pre-service human serum matrix damping fluid in, human serum ratio is 5%, add respectively that concentration is 0,40,80,160ng/ml obtains from the cultivation of duodenum JEG-3, purifying, the PGⅡ antigen of purity >=95%, stir, as PGⅡ multiple spot calibration object.In addition, in pretreated human serum matrix damping fluid, also comprise 1% magnesium sulfate, 4.5% sorbierite, 0.1%Proclin300.
The PGⅡ detection kit of preparing in the present embodiment detects the method for sample, with " eliminating the method for the PGⅡ detection kit detection sample of chyle interference " in embodiment 1.
Two, eliminate the anti-chyle interference performance of the PGⅡ detection kit of chyle interference
According to following method of operating, detect the anti-chyle jamming performance of PGⅡ detection kit:
(1) collect the close human serum sample without haemolysis, jaundice, chyle, turbid phenomenon of measured value of lower scope, mix, this PHS's sample is detected, the value that obtains its PGⅡ is 8.6ng/ml;
(2) according to shown in following table 9, table 10, in PHS, add the chyle chaff interference of variable concentrations gradient;
(3) use respectively certain producer's immunoturbidimetry PGⅡ detection kit (in this test, be called for short kit A) and the PG II detection kit prepared of the present embodiment (in this test, be called for short kit B), to having added the serum sample of different chyle units concentration, carry out blank determination, repeat 3 times, computation of mean values, CV and deviation, deviation range is considered as noiselessly ± 10% with interior, and deviation range surpasses ± 10% and is considered as disturbing.
Table 9 kit A detects the result of adding variable concentrations chyle chaff interference serum sample
Table 10 kit B detects the result of adding variable concentrations chyle chaff interference serum sample
The demonstration of table 9 result, with the range of chyle turbidity from 0 to 8000, the detected value deviation of kit A also increases thereupon, and when chyle turbidity unit is greater than 3000, when detected value and interpolation interference, the deviation of detected value is all greater than 10%, is judged to be existence and disturbs.
The demonstration of table 10 result, with the range of chyle turbidity from 0 to 8000, the variation of the detected value deviation of kit B is less, and when detected value and interpolation interference, the deviation of detected value, all in ± 10%, is judged to be and does not have interference.
This test has shown compares existing immunoturbidimetry method kit, and PGⅡ detection kit of the present invention has obviously improved the antijamming capability to chyle.
Three, eliminate the sensitivity of the PGⅡ detection kit of chyle interference
According to following method of operating, detect the sensitivity of PGⅡ detection kit:
(1) determine that a reagent can detect low concentration value accurately and compare with water is blank, the sample of 1.7ng/ml is selected in this test;
(2) detect 20 water, record absorbance numerical value, calculating mean value (X water) and standard deviation (SD);
(3) sample that 20 concentration of detection are 1.7ng/ml, records absorbance numerical value, calculating mean value (X1.7) and standard deviation (SD);
(4) the absorbance mean value of water adds that 3SD is as absorbance corresponding to lowest detectable limit, because the relation of absorbance and concentration is linear relationship substantially, the concentration that can relatively calculate by the absorbance mean value with 1.7ng/ml sample lowest detectable limit is sensitivity.Formula is 1.7 * (X water+3SD)/X1.7.
The data result of the sensitivity test of the PGⅡ detection kit of preparing in the present embodiment (in this test, being called for short " kit of the present invention ") and contrast agents box (certain producer's immunoturbidimetry PGⅡ detection kit) is as shown in table 11.
In following table, △ A represents the difference of absorbance.
Table 11 kit of the present invention and contrast agents box remolding sensitivity are to data
Sensitivity=2.0 of kit of the present invention * (21.8+3 * 1.3)/690.45=0.07ng/ml.
Sensitivity=2.0 of contrast agents box * (24.35+3 * 11.5)/566.25=0.21ng/ml.
Result shows that the sensitivity of kit of the present invention can reach 0.07ng/ml, is better than contrast agents box.
Kit of the present invention is by being attached to a certain proportion of PG II monoclonal antibody and polyclonal antibody on the latex particle of carboxyl modified with chemical crosslink technique, make on the one hand antigen-antibody binding reaction more rapidly, firmly, by latex agglutination effect, amplify on the other hand the detection progression of reaction, thereby realized the sensitivity raising of kit.From the result of above-mentioned table 11, kit of the present invention highly sensitive in the sensitivity of contrast agents box, visible kit of the present invention has been realized the effect that sensitivity improves.
Four, eliminate the PGⅡ detection kit of chyle interference and the correlativity that enzyme is exempted from method detection kit
PG II detection kit prepared by use the present embodiment is (in this test, be called for short " PG II kit of the present invention ") and the latex immunoturbidimetry PG of our company I detection kit (in this test, be called for short " PG of our company I kit "), adopt Hitachi's 7080 automatic clinical chemistry analyzers, 50 parts of Freshman serum without chyle, haemolysis, jaundice, turbid phenomenon are detected, carry out correlation analysis with the testing result of contrast agents box (the euzymelinked immunosorbent assay (ELISA) propepsin I of Beijing producer, II detection kit are called for short " enzyme is exempted from kit ").PG II kit of the present invention detects according to above-mentioned " PGⅡ detection kit detects the method for sample ", and the PG of our company I kit detects to specifications, and enzyme is exempted from kit and detected according to its instructions, and testing result is as shown in table 12.
In following table, the term of reference that enzyme is exempted from method kit is: PG I (70-200ng/ml); PG II (0-15ng/ml); PG I/PG II >7.5; The term of reference of our company's latex immunoturbidimetry method kit is: PG I (70-240ng/ml); PG II (0-20ng/ml); PG I/PG II >3.
Table 12 kit of the present invention and enzyme are exempted from the clinical data of comparing of method kit
By upper table statistics, can be found out, in this 50 routine fresh patients serum, there are altogether 33 routine negative findingses and 17 routine positive findingses, it is one to one that enzyme is exempted from kit and kit of the present invention in result judgement, and two individual event results and ratio thereof all exist coincideing in clinical tolerance band.Although two kinds of square ratio juris and term of reference have a certain distance, judge from the angle of clinical practice, the correlativity of the two is good, and clinical meaning is remarkable.
Above-described embodiment, the present invention embodiment a kind of more preferably just, the common variation that those skilled in the art carries out within the scope of technical solution of the present invention and replacing all should be included in protection scope of the present invention.
Claims (7)
1. latex immunoturbidimetry detects a detection kit for Serum Pepsinogen II, it is characterized in that, comprises reagent 1, reagent 2 and calibration object;
Described reagent 1 comprises electrolyte, chyle remover, set accelerator, stabilizing agent, surfactant, antiseptic and damping fluid;
Described reagent 2 comprises anti-human PGⅡ antibody coated latex particle, antiseptic and damping fluid;
Described calibration object comprises electrolyte, antiseptic, stabilizing agent, PGⅡ antigen, human serum and damping fluid;
Described electrolyte is selected from inorganic salts;
Described chyle remover, is selected from polyglycol-sulfuric acid chrysanthemum glycan azoviolet of lipase or 1～5%;
Described set accelerator is selected from PEG-4000, PEG-6000, PEG-8000 or sodium dextran sulfate;
Described stabilizing agent is selected from bovine serum albumin(BSA), casein, fishskin gelatin albumen, sorbierite or disodium ethylene diamine tetraacetate;
Described surfactant is selected from tween series, fatty alcohol polyglycol ether, polyoxyethylene phenyl ether, polyoxyethylene laurel ether series;
Described antiseptic is selected from Sodium azide, phenol, P-hydroxybenzoic acid, ethyl-para-hydroxybenzoate, Proclin series;
Described damping fluid is selected from Tris-HCl damping fluid, glycocoll-NaOH damping fluid, phosphate buffer, HEPES-NaOH damping fluid, MOPSO damping fluid;
Described anti-human PGⅡ antibody is monoclonal antibody and the mixing of polyclonal antibody, and wherein monoclonal antibody/how anti-ratio range is 5～10, and antibody type is selected from IgG antibody or IgY antibody;
Described latex particle is polystyrene latex, and diameter range is 150～250nm, and the finishing of latex particle has functional group, and the functional group of modification is selected from carboxyl, amino, hydroxyl, hydrazides or chloromethyl; Anti-human PGⅡ antibody is coated with latex particle surface by chemical crosslink technique, and chemical cross-linking agent is selected a kind of in carbonization imines (EDAC), N-hydroxy-succinamide (NHS), N-hydroxy thiosuccinimide (Sulfo-NHS), hydrazides, isocyanates; Crosslinked damping fluid is selected from MES, MOPSO, MOPS, HEPES damping fluid, and pH is between 6.0～8.0.
2. kit according to claim 1, is characterized in that: described electrolyte is selected from sodium chloride, potassium chloride, magnesium chloride or magnesium sulfate;
Described anti-human PGⅡ antibody is selected from that rabbit is anti-human, goat is anti-human, sheep is anti-human, mouse-anti people, chicken is anti-human, duck is anti-human.
3. kit according to claim 1 and 2, is characterized in that, in described reagent 1, damping fluid surge capability is for regulating pH between 7.0～9.0, and concentration range is 10～100mmol/L; Chyle remover is lipase, concentration range 5～50KU/L; Electrolyte is sodium chloride, and concentration range is 1%～5%; Set accelerator is PEG-6000, and concentration range is 2%～6%; Stabilizing agent is bovine serum albumin, and concentration range is 0.5%～5%; Surfactant is polysorbas20, and concentration range is 0.1%～1.0%; Concentration of preservatives scope is 0.05%～0.1%.
4. kit according to claim 1 and 2, is characterized in that, in described reagent 2, damping fluid surge capability is for regulating pH between 7.0～9.0, and concentration range is 10～100mmol/L; Anti-human PGⅡ antibody is the anti-human IgY antibody of duck; Latex particle finishing carboxyl, chemical cross-linking agent is carbonization imines, and crosslinked damping fluid is HEPES damping fluid, and the antibody latex granule density scope of its sensitization is 0.5～5mg/ml; Concentration of preservatives scope is 0.05%～0.1%.
5. kit according to claim 1 and 2, it is characterized in that, in described calibration object, PGⅡ antigen derive from from human body fluid extraction, purifying and natural PGⅡ antigen, or from the cultivation of duodenum JEG-3, purifying and PGⅡ antigen, its purity >=95%, content is 0～100ng/ml; Human serum ratio is 5%; The concentration range of damping fluid is 10～100mmol/L, and pH is 6.5～8.5; Electrolyte is sodium chloride, and concentration range is 1%～5%; The concentration range of stabilizing agent is 0.5～5%; The concentration range of antiseptic is 0.01%～0.1%.
6. kit according to claim 3, is characterized in that, in described reagent 2, damping fluid surge capability is for regulating pH between 7.0～9.0, and concentration range is 10～100mmol/L; Anti-human PGⅡ antibody is the anti-human IgY antibody of duck; Latex particle finishing carboxyl, chemical cross-linking agent is carbonization imines, and crosslinked damping fluid is HEPES damping fluid, and the antibody latex granule density scope of its sensitization is 0.5～5mg/ml; Concentration of preservatives scope is 0.05%～0.1%.
7. kit according to claim 6, it is characterized in that in described calibration object, PGⅡ antigen derive from from human body fluid extraction, purifying and natural PGⅡ antigen, or from the cultivation of duodenum JEG-3, purifying and PGⅡ antigen, its purity >=95%, content is 0～100ng/ml; Human serum ratio is 5%; The concentration range of damping fluid is 10～100mmol/L, and pH is 6.5～8.5; Electrolyte is sodium chloride, and concentration range is 1%～5%; The concentration range of stabilizing agent is 0.5～5%; The concentration range of antiseptic is 0.01%～0.1%.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1205755A1 (en) * | 2000-05-30 | 2002-05-15 | Iatron Laboratories, Inc. | Immunological latex turbidimetry method and reagent therefor |
| JP2006071574A (en) * | 2004-09-06 | 2006-03-16 | Denka Seiken Co Ltd | Immunoturbidimetry and reagent therefor |
| CN102628864A (en) * | 2011-12-30 | 2012-08-08 | 北京九强生物技术股份有限公司 | Kit for determining heart-type fatty acid binding protein in serum or urine by latex enhanced turbidimetric immunoassay |
| CN102645537A (en) * | 2012-04-26 | 2012-08-22 | 北京美康生物技术研究中心 | Latex enhanced turbidimetric immunoassay kit for diagnosing gastric diseases or gastric cancer, preparation method thereof and application |
| CN102944679A (en) * | 2012-11-15 | 2013-02-27 | 北京康美天鸿生物科技有限公司 | Kit for performing retinol binding protein detection by using latex turbidimetry |
-
2014
- 2014-01-09 CN CN201410010900.4A patent/CN103713140B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1205755A1 (en) * | 2000-05-30 | 2002-05-15 | Iatron Laboratories, Inc. | Immunological latex turbidimetry method and reagent therefor |
| JP2006071574A (en) * | 2004-09-06 | 2006-03-16 | Denka Seiken Co Ltd | Immunoturbidimetry and reagent therefor |
| CN102628864A (en) * | 2011-12-30 | 2012-08-08 | 北京九强生物技术股份有限公司 | Kit for determining heart-type fatty acid binding protein in serum or urine by latex enhanced turbidimetric immunoassay |
| CN102645537A (en) * | 2012-04-26 | 2012-08-22 | 北京美康生物技术研究中心 | Latex enhanced turbidimetric immunoassay kit for diagnosing gastric diseases or gastric cancer, preparation method thereof and application |
| CN102944679A (en) * | 2012-11-15 | 2013-02-27 | 北京康美天鸿生物科技有限公司 | Kit for performing retinol binding protein detection by using latex turbidimetry |
Non-Patent Citations (1)
| Title |
|---|
| 弓福利等: "一种有效去除脂血对血清多种生化指标测定干扰的方法", 《实用医技杂志》, vol. 9, no. 1, 31 January 2002 (2002-01-31), pages 49 * |
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Denomination of invention: A latex immunoturbidimetric pepsinogen II detection kit for eliminating chyle interference Effective date of registration: 20231222 Granted publication date: 20151111 Pledgee: Zhongguancun Branch of Bank of Beijing Co.,Ltd. Pledgor: BEIJING WANTAI DRD CO.,LTD. Registration number: Y2023110000546 |