CN105181977A - Kit for detecting calprotectin in excrement - Google Patents
Kit for detecting calprotectin in excrement Download PDFInfo
- Publication number
- CN105181977A CN105181977A CN201510753055.4A CN201510753055A CN105181977A CN 105181977 A CN105181977 A CN 105181977A CN 201510753055 A CN201510753055 A CN 201510753055A CN 105181977 A CN105181977 A CN 105181977A
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- CN
- China
- Prior art keywords
- calprotectin
- colored particles
- ight soil
- offset plate
- fluorescent particle
- Prior art date
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Classifications
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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
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/06—Gastro-intestinal diseases
- G01N2800/065—Bowel diseases, e.g. Crohn, ulcerative colitis, IBS
Abstract
The invention discloses a kit for detecting calprotectin in excrement. The kit comprises a PVC (Poly Vinyl Chloride) rubber plate, a nitrocellulose membrane, a water-absorbing pad, a colored particle/fluorescent particle compound pad, a sample pad, a detecting line and a quality control line, wherein the nitrocellulose membrane, the water-absorbing pad, the colored particle/fluorescent particle compound pad and the sample pad are arranged on the upper surface of the PVC rubber plate, and the detecting line and the quality control line are arranged on the upper surface of the nitrocellulose membrane. The kit for detecting calprotectin in excrement is used as a dynamic testing kit for domestic or medical institutions and is used for conveniently detecting the content of calprotectin in excrement; when the detecting sensitivity is 15mu g/g, the detecting time is only 3-5 minutes; the operation is simple; the performance is reliable; the cost is low; the sensitivity is high; the detecting time is short; the domestic and overseas blank is filled up.
Description
Technical field
The present invention relates to and a kind ofly detect kit of calprotectin in ight soil and preparation method thereof.
Background technology
Calcium, the zinc-binding protein of calprotectin (Calprotectin) to be a molecular weight be 36kD, the calcium-binding protein heterotrimer that the light chain be heavy chain and a molecular weight of 14kD by two molecular weight being 8kD is connected with covalent bond forms, and every bar chain can in conjunction with two Ca
2+, thus there is thermotolerance and strengthen water-disintegrable characteristic, the compound that its protein structure is made up of 2 subunit S-100A8 and S-100A9.It is distributed in the granulocyte of human body, monocyte, epithelial cell, keratinocyte and various tissue and body fluid, is protective protein.
Calprotectin is a kind of acute inflammation mark, derive from the calcic albumen of neutrophil leucocyte and macrophage, its expression has tissue or cell-specific, is a kind of calbindin of heterozygosity, there is the activity of antiprotease, and there is chelated zinc ions ability and there is heat impedance.Calprotectin has antimicrobial, the various biological function such as immunity moderation, antiproliferative, transmission of signal, raises under many inflammatory conditions.When inflammatory reaction, it is an aggressive process that neutrophil leucocyte enters tissue, it is under the effect of chemotactic factor (CF), by consumed energy, in amoeba sample sex change campaign, get out vascular wall and arrive areas of inflammation, differentiation and maturation is macrophage, this part macrophage, as the effector cell of inflammation, discharges various inflammatory factor, carries out engulfing playing its biological function simultaneously.In neutrophil leucocyte, calprotectin is mainly present in the tenuigenin outside lysozyme, accounts for greatly 5% of neutrophil leucocyte Tot Prot.When neutrophil activation, the calprotectin of phosphorylation is displaced on cell membrane, as the Reverse transcriptase substrate of enzyme, and the enzyme (casein kinase II and topoisomerase etc.) that on cell proliferation can be suppressed to play an important role.Calprotectin can also the generation of immune stimulatory globulin, the activation of chemotactic factor (CF) and neutrophil leucocyte immobilisation factor activation.
Inflammatory bowel disease (IBD) is for involving a kind of idiopathic bowl inflammatory diseases of ileum, rectum, colon.Clinical manifestation diarrhoea, stomachache, even can have blood in stool.Inflammatory bowel disease is the chronic nonspecific bowl inflammatory diseases that a kind of cause of disease is still not clear, and comprises ulcerative colitis (UC) and Crohn disease (CD).The former is a kind of chronic nonspecific colitis disease, and pathology mainly involves colonic mucosa and submucosa, and scope is many from section colon far away, can drive in the wrong direction to nearly section and develop, even involve total colectomy and terminal ileum, distribute in continuity, clinical main manifestations is diarrhoea, suffers from abdominal pain and the dense bloody stool of mucus.The latter is a kind of CGI, and pathology can involve each position of intestines and stomach, and based on terminal ileum and contiguous colon thereof, in transmural inflammation, how in segmental, asymmetry distribution, clinical main manifestations is stomachache, diarrhoea, fistula, anus pathology lamp.Both all can merge constitutional symptom in various degree.
Research finds, the calprotectin levels in the ight soil of inflammatory bowel disease (IBD) patient apparently higher than colorectal cancer patients and IBS (IBS) patient, and is proportionate with patient's lesion degree.In diagnosis of colon cancer process, fecal occult blood checks that (FOBT) is a kind of method of extensive utilization, it judges disease of digestive tract by the haemoglobin, protoheme etc. in detection ight soil, therefore can not find without hemorrhage pathology, and colon cancer can infiltrate to perienchyma, produce inflammatory cell, cause the calprotectin of certain level to express, therefore in ight soil, the calprotectin content normal person that compares increases to some extent.For IBS (IBS) patient, clinician mainly makes tentative diagnosis according to medical history and Clinical symptoms, carry out corresponding auxiliary examination simultaneously, as Sigmoidoscope and clysis with barium etc., except just can show that IBS (IBS) is diagnosed after organic bowel diseases, carry out hauling type examination and become the main method that IBS (IBS) diagnoses, add the misery of medical expense and patient to a certain extent.And calprotectin is mainly expressed in the macrophage matter of inflammatory cell as neutrophil leucocyte and activation, so IBS (IBS) is without inflammatory reaction, calprotectin levels is lower.
Therefore, detecting calprotectin in ight soil can as differentiation inflammatory bowel disease, colon cancer, IBS, and is used for monitoring the activity of inflammatory bowel disease, has great importance to disease palindromia, result for the treatment of.At present for detecting the method mainly enzyme linked immunosorbent assay (ELISA) of calprotectin, although ELISA method sensitivity is higher, but operate more loaded down with trivial details, detect consuming time long, and checkout equipment is not easy to more greatly mobile, therefore need one easy and simple to handle, detect fast method to detect the content of calprotectin.
Summary of the invention
For the deficiencies in the prior art, the invention provides a kind of simple to operate, dependable performance, kit with low cost.
To achieve these goals, the technical solution used in the present invention is as follows:
A kind of kit detecting calprotectin in ight soil, comprise PVC offset plate, be arranged at the nitrocellulose filter of PVC offset plate upper surface, adsorptive pads, colored particles/fluorescent particle bond pad, sample pad, and be arranged at detection line and the nature controlling line of nitrocellulose filter upper surface.
Particularly, described nitrocellulose filter is positioned in the middle part of PVC offset plate; Described adsorptive pads is positioned at wherein one end of PVC offset plate, and one end of adsorptive pads is overlapped in nitrocellulose filter upper surface; Described colored particles/fluorescent particle bond pad is positioned at the other end of PVC offset plate, and one end of colored particles/fluorescent particle bond pad is overlapped in nitrocellulose filter upper surface; Described sample pad is positioned at the end of the PVC offset plate other end, and one end of sample pad is overlapped in colored particles/English hat particulate binding layer upper surface.
Particularly, the width of described sample pad, colored particles/fluorescent particle bond pad, nitrocellulose filter, adsorptive pads is equal with the width of PVC offset plate, and described sample pad, colored particles/fluorescent particle bond pad, nitrocellulose filter, adsorptive pads all attach to PVC offset plate upper surface.
Particularly, described sample pad by glass fibre element film through carrying out processing containing the sample pad treating fluid of sodium tetraborate, casein sodium, polyvinylpyrrolidone, Brij and under 37 DEG C of conditions dry 4 hours and obtain.
Particularly; described colored particles/fluorescent particle bond pad by dacron film through carrying out processing containing the treating fluid of sodium tetraborate, casein sodium, Tween-20 and under 37 DEG C of conditions after dry 4 hours, spraying colored particles or fluorescent particle-mouse anti-human calprotectin monoclonal antibody bond and obtain.
Particularly, described detection line is the detection line being coated with immobilised mouse anti-human calprotectin monoclonal antibody.
Particularly, described nature controlling line is be coated with the how anti-nature controlling line of sheep anti-mouse igg.
Particularly, described colored particles/fluorescent particle is collaurum, latex beads or fluorescent microsphere.
Particularly, described adsorptive pads is absorbent filter.
In addition, the present invention also comprises the cartridge for holding PVC offset plate, and is provided with well in cartridge.
When the present invention detects, specifically the ight soil extract liquid-transfering gun after sample treatment solution process is drawn 100 μ l and add kit well, react 4 minutes under normal temperature, observing response result;
Or ight soil extract is placed in stool preprocessing system, automatically extract is added kit well by system, react 3 ~ 5 minutes under normal temperature, then observing response result.
As calprotectin content >=15 μ g/g in sample, testing result will be positive.
The present invention compared with prior art, has the following advantages and beneficial effect:
The present invention is simple to operate, dependable performance, with low cost.
Accompanying drawing explanation
Fig. 1 is structural representation of the present invention.
Fig. 2 is that yin and yang attribute of the present invention differentiates schematic diagram.
Wherein, mark corresponding parts name in accompanying drawing to be called: 1-PVC offset plate, 2-nitrocellulose filter, 3-adsorptive pads, 4-colored particles/fluorescent particle bond pad, 5-sample pad, 6-detection line, 7-nature controlling line.
Embodiment
Below in conjunction with drawings and Examples, the invention will be further described, and embodiments of the present invention include but not limited to the following example.
Embodiment
As shown in Figure 1, a kind of kit detecting calprotectin in ight soil, comprise PVC offset plate 1, be arranged at the nitrocellulose filter 2 of PVC offset plate upper surface, adsorptive pads 3, colored particles/fluorescent particle bond pad 4, sample pad 5, and be arranged at detection line 6 and the nature controlling line 7 of nitrocellulose filter upper surface.
Particularly, described nitrocellulose filter is positioned in the middle part of PVC offset plate; Described adsorptive pads is positioned at wherein one end of PVC offset plate, and one end of adsorptive pads is overlapped in nitrocellulose filter upper surface; Described colored particles/fluorescent particle bond pad is positioned at the other end of PVC offset plate, and one end of colored particles/fluorescent particle bond pad is overlapped in nitrocellulose filter upper surface; Described sample pad is positioned at the end of the PVC offset plate other end, and one end of sample pad is overlapped in colored particles/fluorescent particle binding layer upper surface.
Particularly, the width of described sample pad, colored particles/fluorescent particle bond pad, nitrocellulose filter, adsorptive pads is equal with the width of PVC offset plate, and described sample pad, colored particles/fluorescent particle bond pad, nitrocellulose filter, adsorptive pads all attach to PVC offset plate upper surface.
Particularly, described sample pad by glass fibre element film through carrying out processing containing the sample pad treating fluid of sodium tetraborate, casein sodium, polyvinylpyrrolidone, Brij and under 37 DEG C of conditions dry 4 hours and obtain.Its effect be in order to tackle bulky grain in sample and as detection system buffering to ensure that in reaction system, calprotectin and mouse anti-human calprotectin monoclonal antibody carry out specific combination.
Particularly; described colored particles/fluorescent particle bond pad by dacron film through carrying out processing containing the treating fluid of sodium tetraborate, casein sodium, Tween-20 and under 37 DEG C of conditions after dry 4 hours; spraying colored particles or fluorescent particle-mouse anti-human calprotectin monoclonal antibody bond and obtaining, can the sensitivity of guarantee reagent box and specificity by spraying process.
Particularly, described detection line is the detection line being coated with immobilised mouse anti-human calprotectin monoclonal antibody;
Particularly, described nature controlling line is be coated with the how anti-nature controlling line of sheep anti-mouse igg.For reacting shown coloured band or fluorescent bands is the standard determining whether enough samples and chromatography process.
Particularly, described colored particles/fluorescent particle is collaurum, latex beads or fluorescent microsphere.
Particularly, described adsorptive pads is absorbent filter.Sample Tomography Velocity can be accelerated, reach the object detecting calprotectin in ight soil fast.
In addition, the present invention also comprises the cartridge for holding PVC offset plate, and is provided with well in cartridge.This cartridge is used for above-mentioned substance to hold in the inner, and is fixed, thus conveniently detects.
During use, when adding sample in the well of kit, the ight soil extract of the people namely after sample treatment solution process, this sample moves to colored particles/fluorescent particle bond pad by sample pad, calprotectin in sample can be combined with colored particles/fluorescent particle-mouse anti-human calprotectin monoclonal antibody protein conjugates specifically and form colored particles or fluorescent particle-mouse anti-human calprotectin antibody-calprotectin, on nitrocellulose filter, the double-antibody sandwich bond of detection line and its formation colored particles or the fluorescent particle-mouse anti-human calprotectin antibody-calprotectin-mouse anti-human calprotectin antibody being coated with mouse anti-human calprotectin antibody is being moved across by capillarity, and present and have band or fluorescent bands, result is positive, if not containing calprotectin in sample, then detection line place will not have coloured band or fluorescent bands, result is negative.No matter whether calprotectin is present in sample, and a coloured band or fluorescent bands all can appear at nature controlling line place.The coloured band that nature controlling line place shows or fluorescent bands have determined whether enough samples and the whether normal standard of chromatography process.
The interpretation of kit of the present invention, as shown in Figure 2, when the calprotectin content in sample is less than 15 μ g/g, the view window region of kit is by the nature controlling line of appearance coloured band or fluorescent bands, and testing result is negative.When the calprotectin content in sample is equal to or greater than 15 μ g/g, the view window region of kit is by appearance coloured band or the detection line of fluorescent bands and the nature controlling line of a coloured band or fluorescent bands, and testing result is not positive.When nature controlling line does not appear in the view window region of kit, no matter detect and whether occur detection line, it is invalid that the interpretation of this kit is.In Fig. 2, first kit testing result is positive from left to right, and second is the weak positive, and the 3rd is negative, and the 4th and the 5th is invalid.
What deserves to be explained is, the present invention, as a kind of family expenses or medical institutions detection of dynamic reagent, can detect calprotectin content in ight soil easily.When its detection sensitivity is 15 μ g/g, detection time only needs 3 ~ 5 minutes, and it is simple to operate, dependable performance, with low cost, highly sensitive, detection time is short, has filled up domestic and international blank.
According to above-described embodiment, just the present invention can be realized well.What deserves to be explained is; under prerequisite based on said structure design, for solving same technical matters, even if some making on the invention are without substantial change or polishing; the essence of the technical scheme adopted is still the same with the present invention, therefore it also should in protection scope of the present invention.
Claims (10)
1. one kind is detected the kit of calprotectin in ight soil, it is characterized in that, comprise PVC offset plate (1), be arranged at the nitrocellulose filter (2) of PVC offset plate upper surface, adsorptive pads (3), colored particles/fluorescent particle bond pad (4), sample pad (5), and be arranged at detection line (6) and the nature controlling line (7) of nitrocellulose filter upper surface.
2. a kind of kit detecting calprotectin in ight soil according to claim 1, is characterized in that, described nitrocellulose filter is positioned in the middle part of PVC offset plate; Described adsorptive pads is positioned at wherein one end of PVC offset plate, and one end of adsorptive pads is overlapped in nitrocellulose filter upper surface; Described colored particles/fluorescent particle bond pad is positioned at the other end of PVC offset plate, and one end of colored particles/fluorescent particle bond pad is overlapped in nitrocellulose filter upper surface; Described sample pad is positioned at the end of the PVC offset plate other end, and one end of sample pad is overlapped in colored particles/fluorescent particle binding layer upper surface.
3. a kind of kit detecting calprotectin in ight soil according to claim 1, it is characterized in that, the width of described sample pad, colored particles/fluorescent particle bond pad, nitrocellulose filter, adsorptive pads is equal with the width of PVC offset plate, and described sample pad, colored particles/fluorescent particle bond pad, nitrocellulose filter, adsorptive pads all attach to PVC offset plate upper surface.
4. a kind of kit detecting calprotectin in ight soil according to claim 1, it is characterized in that, described sample pad by glass fibre element film through carrying out processing containing the sample pad treating fluid of sodium tetraborate, casein sodium, polyvinylpyrrolidone, Brij and under 37 DEG C of conditions dry 4 hours and obtain.
5. a kind of kit detecting calprotectin in ight soil according to claim 1; it is characterized in that; described colored particles/fluorescent particle bond pad by dacron film through carrying out processing containing the treating fluid of sodium tetraborate, casein sodium, Tween-20 and under 37 DEG C of conditions after dry 4 hours, spraying colored particles or fluorescent particle-mouse anti-human calprotectin monoclonal antibody bond and obtain.
6. a kind of kit detecting calprotectin in ight soil according to claim 1, is characterized in that, described detection line is the detection line being coated with immobilised mouse anti-human calprotectin monoclonal antibody.
7. a kind of kit detecting calprotectin in ight soil according to claim 1, is characterized in that, described nature controlling line is be coated with the how anti-nature controlling line of sheep anti-mouse igg.
8. a kind of kit detecting calprotectin in ight soil according to claim 1, is characterized in that, described colored particles/fluorescent particle is collaurum, latex beads or fluorescent microsphere.
9. a kind of kit detecting calprotectin in ight soil according to claim 1, is characterized in that, described adsorptive pads is absorbent filter.
10. a kind of kit detecting calprotectin in ight soil according to claim 1, is characterized in that, also comprises the cartridge for holding PVC offset plate, and be provided with well in cartridge.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201510753055.4A CN105181977A (en) | 2015-11-05 | 2015-11-05 | Kit for detecting calprotectin in excrement |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201510753055.4A CN105181977A (en) | 2015-11-05 | 2015-11-05 | Kit for detecting calprotectin in excrement |
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| CN105181977A true CN105181977A (en) | 2015-12-23 |
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| CN201510753055.4A Pending CN105181977A (en) | 2015-11-05 | 2015-11-05 | Kit for detecting calprotectin in excrement |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105717308A (en) * | 2016-03-24 | 2016-06-29 | 泰州海路生物技术有限公司 | Immunochromatography kit for fast and quantitatively detecting fecal lactoferrin |
| CN105785041A (en) * | 2016-04-05 | 2016-07-20 | 付国亮 | Test strip for quantitatively detecting calprotectin, preparation method thereof and determining method for calprotectin concentration |
| CN105842459A (en) * | 2016-03-24 | 2016-08-10 | 泰州海路生物技术有限公司 | Immuno-chromatography kit for quickly and quantitatively detecting calprotectin in excrement |
| CN107698675A (en) * | 2017-07-12 | 2018-02-16 | 武汉博百欧生物科技有限公司 | Separate and purify the method and kit of calprotectin |
| CN108614113A (en) * | 2018-06-07 | 2018-10-02 | 河南百奥生物工程有限公司 | A kind for the treatment of fluid and preprocess method of colloidal gold immune chromatography test gold-labelled pad |
| CN109813917A (en) * | 2019-02-22 | 2019-05-28 | 山东爱维德生物科技有限公司 | A kind of calprotectin detection kit and its test method |
| CN110244062A (en) * | 2019-07-18 | 2019-09-17 | 珠海市医友生物科技有限公司 | A kind of excrement calprotectin or lactoferrin combined detection reagent and preparation method thereof |
| CN112578125A (en) * | 2019-09-27 | 2021-03-30 | 成都中医药大学 | Application of reagent for detecting content of calprotectin in preparation of ovarian lesion screening kit |
| WO2021057988A1 (en) * | 2019-09-27 | 2021-04-01 | 廖睿 | Use of reagent for detecting content of faecal calprotectin in preparation of kit for screening fallopian tube lesions |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105717308A (en) * | 2016-03-24 | 2016-06-29 | 泰州海路生物技术有限公司 | Immunochromatography kit for fast and quantitatively detecting fecal lactoferrin |
| CN105842459A (en) * | 2016-03-24 | 2016-08-10 | 泰州海路生物技术有限公司 | Immuno-chromatography kit for quickly and quantitatively detecting calprotectin in excrement |
| CN105785041A (en) * | 2016-04-05 | 2016-07-20 | 付国亮 | Test strip for quantitatively detecting calprotectin, preparation method thereof and determining method for calprotectin concentration |
| CN107698675A (en) * | 2017-07-12 | 2018-02-16 | 武汉博百欧生物科技有限公司 | Separate and purify the method and kit of calprotectin |
| CN108614113A (en) * | 2018-06-07 | 2018-10-02 | 河南百奥生物工程有限公司 | A kind for the treatment of fluid and preprocess method of colloidal gold immune chromatography test gold-labelled pad |
| CN109813917A (en) * | 2019-02-22 | 2019-05-28 | 山东爱维德生物科技有限公司 | A kind of calprotectin detection kit and its test method |
| CN110244062A (en) * | 2019-07-18 | 2019-09-17 | 珠海市医友生物科技有限公司 | A kind of excrement calprotectin or lactoferrin combined detection reagent and preparation method thereof |
| CN112578125A (en) * | 2019-09-27 | 2021-03-30 | 成都中医药大学 | Application of reagent for detecting content of calprotectin in preparation of ovarian lesion screening kit |
| WO2021057988A1 (en) * | 2019-09-27 | 2021-04-01 | 廖睿 | Use of reagent for detecting content of faecal calprotectin in preparation of kit for screening fallopian tube lesions |
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Application publication date: 20151223 |