CN104049084A - Cystine protease inhibitor C detection kit - Google Patents
Cystine protease inhibitor C detection kit Download PDFInfo
- Publication number
- CN104049084A CN104049084A CN201310075686.6A CN201310075686A CN104049084A CN 104049084 A CN104049084 A CN 104049084A CN 201310075686 A CN201310075686 A CN 201310075686A CN 104049084 A CN104049084 A CN 104049084A
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- China
- Prior art keywords
- reagent
- protease inhibitor
- concentration range
- kit
- present
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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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/544—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being organic
-
- 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
- G01N2333/81—Protease inhibitors
- G01N2333/8107—Endopeptidase (E.C. 3.4.21-99) inhibitors
- G01N2333/8139—Cysteine protease (E.C. 3.4.22) inhibitors, e.g. cystatin
Abstract
The invention discloses a cystine protease inhibitor C detection kit which includes a reagent 1 and a reagent 2. The reagent 1 is a glycine buffer solution with a concentration range being 0.01-5 mol/L. The reagent 2 is an anti-human CYS-C antibody latex particle with a concentration range being 100-500 [mu]l/mL. The kit has advantages of rapid reaction, high sensitivity, good accuracy and a long stability time.
Description
Technical field
The present invention relates to a kind of cysteine protease inhibitor C test kit in serum of measuring, be specifically related to a kind of Immunoturbidimetry that uses and measure cysteine protease inhibitor C test kit in human serum.
Background technology
Cystine protease inhibitors (CYS-C) is a kind of desirable endogenous mark that reflects that glomerular filtration rate(GFR changes.In detection of glomeruli filtration function test, serum creatinine, urea and endogenous creatinine clearance rate are the most frequently used indexs.Because above index remains many deficiencies, old friends are finding the index that new reflection glomerular filtration rate(GFR changes always.Bladder chalone C in circulation is only eliminated through glomerular filtration, and bladder chalone C can provide early diagnosis to early stage in ephrosis, kidney transplant, diabetes, ephrosis etc.; Owing to not being subject to the impact of the factor such as age, muscle mass, to old man, children, pregnant woman's diagnostic result more accurately and reliably, its diagnosis effect will be obviously due to serum creatinine simultaneously.
Summary of the invention
The object of invention is to overcome the defect that prior art exists, and provides a kind of reaction velocity fast, and sensitivity is strong, and accuracy is high, the cysteine protease inhibitor C test kit that stationary phase is long.
The technical scheme that realizes the object of the invention is: a kind of cysteine protease inhibitor C test kit is provided, the liquid double reagent that this kit is made up of reagent 1 and reagent 2, wherein said reagent 1 is glycine buffer, its concentration range is: 0.01 ~ 5mol/L, reagent 2 is anti-human CYS-C antibody latex particle, and its concentration range is: 100 ~ 500 μ l/mL.
As preferably, cysteine protease inhibitor C test kit of the present invention, described reagent 1 is glycine buffer, its concentration range is: 0.05 ~ 2mol/L, reagent 2 is anti-human CYS-C antibody latex particle, its concentration range is: 200 ~ 400 μ l/mL.
As most preferably, cysteine protease inhibitor C test kit of the present invention, described reagent 1 is glycine buffer, its concentration range is: 0.1mol/L, reagent 2 is anti-human CYS-C antibody latex particle, its concentration range is: 300 μ l/mL.
Cysteine protease inhibitor C test kit of the present invention, the volume ratio of described reagent 1 and reagent 2 is: 4:1.
Cysteine protease inhibitor C test kit of the present invention, the anti-human CYS-C antibody latex particle in described reagent 2 is the coated latex particle of monoclonal antibody, diameter is 100nm.
Raw material in cysteine protease inhibitor C test kit of the present invention in reagent 1 and reagent 2 is selected according to being:
Glycine buffer is stable damping fluid, is beneficial to the dispersion of various compositions in reagent, can improve the stability of reagent.The coated latex particle of monoclonal antibody that anti-human CYS-C antibody latex particle adopts, latex particle adopts nanometer technology, and diameter reaches 100nm, and reaction velocity is accelerated, and improves sensitivity simultaneously.
The collocation method of kit reagent 1 of the present invention and reagent 2 is conventional method, and reagent 1 and reagent 2 mix and stir evenly separately after adding respectively distilled water.
Kit of the present invention is measured Cystatin C by following method, anti-human CYS-C antibody latex particle generation agglutinating reaction in Cystatin C (CYS-C) in sample and reagent, agglutinating reaction forms antigen antibody complex and produces turbidity, and its turbidity height is directly proportional to CYS-C in sample in the time that a certain amount of antibody exists.By measuring the absorbance of specific wavelength, can calculate the content of CYS-C in serum with reference to calibration curve.
The assay method of applying Cystatin C in cysteine protease inhibitor C test kit mensuration sample of the present invention is as follows: sample reagent adding 1 mixes, after 30 ~ 40 DEG C of reaction 3 ~ 5min, read absorbance A 1, add reagent 2 to mix, after 30 ~ 40 DEG C of reaction 3 ~ 5min, read absorbance A 2, Δ A=A2-A1.Wherein amount of samples 8 μ L, reagent 1 consumption 200-300 μ L, reagent 2 consumption 30-110 μ L.
Apply cysteine protease inhibitor C test kit of the present invention and measure the content of Cystatin C in sample, computing formula is as follows:
Cystatin C=sample absorbance (A2-A1) × concentration of standard solution/titer absorbance Δ A0,
The present invention has positive effect: with existing cysteine protease inhibitor C test kit, reaction velocity of the present invention is fast, highly sensitive, and the stable reagent phase is long, is applicable to automatic clinical chemistry analyzer, has larger clinical value.
Brief description of the drawings
For content of the present invention is more easily expressly understood, according to specific embodiment also by reference to the accompanying drawings, the present invention is further detailed explanation, wherein below
Fig. 1 is the real-time reflection curve of kit measurement Cystatin C of the present invention.
Fig. 2 is the real-time reflection curve that contrast agent box is measured Cystatin C.
In Fig. 1, Fig. 2, horizontal ordinate is reading point, and ordinate is absorbance A.
Embodiment
Below in conjunction with the drawings and specific embodiments, the present invention is described in further detail, but the present invention is far not limited only to following examples.
Embodiment 1
Reagent 1(R1) component and concentration range thereof:
R1 glycine buffer 0.01mol/L
Wherein reagent 2(R2) component and concentration range thereof:
The anti-human CYS-C antibody latex of R2 particle 100 μ l/mL
The collocation method of reagent 1 and reagent 2 is conventional method, and the group described in reagent 1 and reagent 2 is mixed and stirred evenly separately after adding respectively distilled water.
Embodiment 2
Reagent 1(R1) and concentration range:
R1 glycine buffer 5mol/L
Wherein reagent 2(R2) and concentration range:
The anti-human CYS-C antibody latex of R2 particle 500 μ l/mL
The preparation method of embodiment 2 kits is with embodiment 1.
Embodiment 3
Reagent 1(R1) and concentration range:
R1 glycine buffer 0.05 mol/L
Wherein reagent 2(R2) and concentration range:
The anti-human CYS-C antibody latex of R2 particle 200 μ l/mL
The preparation method of embodiment 3 kits is with embodiment 1.
Embodiment 4
Reagent 1(R1) and concentration range:
R1 glycine buffer 2mol/L
Wherein reagent 2(R2) and concentration range:
The anti-human CYS-C antibody latex of R2 particle 400 μ l/mL
Following examples.
The preparation method of embodiment 4 kits is with embodiment 1.
Embodiment 5
Reagent 1(R1) and concentration range:
R1 glycine buffer 0.1mol/L
Wherein reagent 2(R2) and concentration range:
The anti-human CYS-C antibody latex of R2 particle 300 μ l/mL
The preparation method of embodiment 5 kits is with embodiment 1.
The assay method of applying Cystatin C in cysteine protease inhibitor C test kit mensuration sample of the present invention is as follows: sample reagent adding 1 mixes, after 30 ~ 40 DEG C of reaction 3 ~ 5min, read absorbance A 1, add reagent 2 to mix, after 30 ~ 40 DEG C of reaction 3 ~ 5min, read absorbance A 2, wherein amount of samples 8 μ L, reagent 1 consumption 200-300 μ L, reagent 2 consumption 30-110 μ L.
According to Cystatin C=sample absorbance Δ A × concentration of standard solution/titer absorbance Δ A0, draw the content of Cystatin C.
Below in conjunction with Fig. 1, Fig. 2 and form, the kit contrasting and the performance of kit of the present invention are compared.By relatively these two kinds of kits of impact of response curve, sensitivity, the range of linearity, accuracy and various chaff interferences etc.
Comparative example
Comparative example cysteine protease inhibitor C test kit, its composition and formula are
Reagent 1: damping fluid 12mmol/L
Reagent 2: the antibody 150mmol/L that collaurum is coated
Kit of the present invention: reagent 1 and reagent 2.
1, the comparison of the range of linearity of two kinds of kits:
Preparation 0,1.0,2.0,3.0,4.0,5.0,6.0,7.0,8.0,9.0,10.0mg/L, Cystatin C standard solution.Measure its concentration with kit of the present invention and contrast agent box respectively, the results are shown in Table 1.
Table 1 kit of the present invention and contrast agent box are measured range of linearity contrast
The range of linearity of kit of the present invention is compared with contrast agent box, and in the time that Cystatin C concentration is greater than 6.0mg/L, contrast agent box can not detect, and the range of linearity is too small, illustrates that kit of the present invention has linear test specification large.
2, the comparison of the sensitivity of two kinds of kits:
With the quality-control product of existing kit and kit measurement of the present invention 1.88 mg/L, poor (A) reading of absorbance is as follows:
| Absorbance | A 1 | A 2 | ?A=A 2-A 1 |
| Kit of the present invention | 231 | 2570 | 2339 |
| Contrast agent box | 258 | 1850 | 1492 |
Kit of the present invention absorbance numerical value when comparing the same sample of kit measurement is large, illustrates that the present invention is highly sensitive.
3, the comparison of two kinds of kit effect phase stability:
The expired kit that exceedes 1.5 years with the kit contrasting and effect phase carries out contrast test, the results are shown in Table 2.
Table 2 is imitated phase stability test comparing
The precision of kit of the present invention, highly sensitive in contrast agent box as can be seen from the above table, effect phase stability is also greater than contrast agent box, still can use after exceeding 1.5 years.
Have reaction velocity from the known cysteine protease inhibitor C test kit of the present invention of above-described embodiment faster, sensitivity is higher, the advantage that the stable reagent phase is longer.
Above-described specific embodiment; object of the present invention, technical scheme and beneficial effect are further described; institute is understood that; the foregoing is only specific embodiments of the invention; be not limited to the present invention; within the spirit and principles in the present invention all, any amendment of making, be equal to replacement, improvement etc., within all should being included in protection scope of the present invention.
Claims (5)
1. a cysteine protease inhibitor C test kit, is characterized in that: the liquid-type double reagent that this kit is made up of reagent 1 and reagent 2, and described reagent 1 is glycine buffer, its concentration range is: 0.01 ~ 5mol/L,
Reagent 2 is anti-human CYS-C antibody latex particle, and its concentration range is: 100 ~ 500 μ l/mL.
2. cysteine protease inhibitor C test kit according to claim 1, it is characterized in that: described reagent 1 is glycine buffer, its concentration range is: 0.05 ~ 2mol/L, and reagent 2 is anti-human CYS-C antibody latex particle, its concentration range is: 200 ~ 400 μ l/mL.
3. cysteine protease inhibitor C test kit according to claim 2, it is characterized in that: described reagent 1 is glycine buffer, its concentration range is: 0.1mol/L, and reagent 2 is anti-human CYS-C antibody latex particle, its concentration range is: 300 μ l/mL.
4. according to the cysteine protease inhibitor C test kit described in claim 1,2 or 3, it is characterized in that: the reagents ratio of described reagent 1 and reagent 2 is: 4:1.
5. according to the cysteine protease inhibitor C test kit described in claim 1,2 or 3, it is characterized in that: in described reagent 2, anti-human CYS-C antibody latex particle is the coated latex particle of monoclonal antibody, and diameter is 100nm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310075686.6A CN104049084A (en) | 2013-03-11 | 2013-03-11 | Cystine protease inhibitor C detection kit |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310075686.6A CN104049084A (en) | 2013-03-11 | 2013-03-11 | Cystine protease inhibitor C detection kit |
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| CN104049084A true CN104049084A (en) | 2014-09-17 |
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| CN201310075686.6A Pending CN104049084A (en) | 2013-03-11 | 2013-03-11 | Cystine protease inhibitor C detection kit |
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Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2000193662A (en) * | 1998-12-25 | 2000-07-14 | Tomakomai Rinsho Kensa Center:Kk | Measuring reagent for cystatin c in urine, diagnostic method and kit |
| CN101377492A (en) * | 2007-08-29 | 2009-03-04 | 北京九强生物技术有限公司 | Bladder chalone C determining reagent kit |
| CN101413001A (en) * | 2008-11-28 | 2009-04-22 | 四川省迈克科技有限责任公司 | Recombinant human cystatin C genes, and expression and use thereof |
| CN102095872A (en) * | 2010-11-08 | 2011-06-15 | 武汉生之源生物科技有限公司 | Enhanced cystatin C assay kit based on large-particle-size latex |
| CN102279269A (en) * | 2011-04-12 | 2011-12-14 | 王贤俊 | Preparation method of cystatin C detection kit |
| CN102353770A (en) * | 2011-06-03 | 2012-02-15 | 宁波美康生物科技有限公司 | Detection kit for cystine protease inhibitor C |
| CN102590497A (en) * | 2012-01-13 | 2012-07-18 | 宁波美康生物科技股份有限公司 | Cysteine protease inhibitor C test kit |
| CN102636653A (en) * | 2012-04-19 | 2012-08-15 | 上海蓝怡科技有限公司 | Compounded latex particle-enveloped cystatin C detection kit |
| CN102749458A (en) * | 2012-07-27 | 2012-10-24 | 北京恩济和生物科技有限公司 | Cystatin C detection kit and preparing method thereof |
| CN102879569A (en) * | 2012-09-18 | 2013-01-16 | 苏州照康生物技术有限公司 | Immunoturbidimetry kit for detecting cysteine proteinase inhibitor C and preparation method thereof |
-
2013
- 2013-03-11 CN CN201310075686.6A patent/CN104049084A/en active Pending
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2000193662A (en) * | 1998-12-25 | 2000-07-14 | Tomakomai Rinsho Kensa Center:Kk | Measuring reagent for cystatin c in urine, diagnostic method and kit |
| CN101377492A (en) * | 2007-08-29 | 2009-03-04 | 北京九强生物技术有限公司 | Bladder chalone C determining reagent kit |
| CN101413001A (en) * | 2008-11-28 | 2009-04-22 | 四川省迈克科技有限责任公司 | Recombinant human cystatin C genes, and expression and use thereof |
| CN102095872A (en) * | 2010-11-08 | 2011-06-15 | 武汉生之源生物科技有限公司 | Enhanced cystatin C assay kit based on large-particle-size latex |
| CN102279269A (en) * | 2011-04-12 | 2011-12-14 | 王贤俊 | Preparation method of cystatin C detection kit |
| CN102353770A (en) * | 2011-06-03 | 2012-02-15 | 宁波美康生物科技有限公司 | Detection kit for cystine protease inhibitor C |
| CN102590497A (en) * | 2012-01-13 | 2012-07-18 | 宁波美康生物科技股份有限公司 | Cysteine protease inhibitor C test kit |
| CN102636653A (en) * | 2012-04-19 | 2012-08-15 | 上海蓝怡科技有限公司 | Compounded latex particle-enveloped cystatin C detection kit |
| CN102749458A (en) * | 2012-07-27 | 2012-10-24 | 北京恩济和生物科技有限公司 | Cystatin C detection kit and preparing method thereof |
| CN102879569A (en) * | 2012-09-18 | 2013-01-16 | 苏州照康生物技术有限公司 | Immunoturbidimetry kit for detecting cysteine proteinase inhibitor C and preparation method thereof |
Non-Patent Citations (4)
| Title |
|---|
| 于德军等: "乳胶增强免疫比浊法测定血清胱抑素C", 《检验医学与临床》 * |
| 谭功军: "胱抑素C试剂盒的研发及临床应用", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
| 陈婷梅: "人胱抑素C抗体制备和检测方法建立及其方法学评价", 《中国博士学位论文全文数据库》 * |
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Application publication date: 20140917 |