CN106370648A - Detection kit with stable unsaturated iron-binding capacity - Google Patents
Detection kit with stable unsaturated iron-binding capacity Download PDFInfo
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- CN106370648A CN106370648A CN201610742667.8A CN201610742667A CN106370648A CN 106370648 A CN106370648 A CN 106370648A CN 201610742667 A CN201610742667 A CN 201610742667A CN 106370648 A CN106370648 A CN 106370648A
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- Prior art keywords
- reagent
- binding capacity
- unsaturated iron
- iron
- buffer
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N2021/3185—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry typically monochromatic or band-limited
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Abstract
The invention discloses a detection kit with stable unsaturated iron-binding capacity, belonging to the technical field of clinical in-vitro detection reagents. The kit comprises a reagent R1 and a reagent R2. The buffer solution of the reagent R1 is replaced with an AMP (adenosine monophosphate) buffer solution with the pH value of 8.6, and ascorbic acid is added into the reagent R2, thereby enhancing the stability of the kit; and the kit has favorable linear dependence and high reagent accuracy, and is beneficial to further popularization and application in the market.
Description
Technical field
The present invention relates to clinical vitro detection reagent technique field, particularly to a kind of unsaturated iron-binding capacity detectable
Box.
Background technology
Normal human's iron-holder about 4g about, wherein 2/3 is present in the middle of endoerythrocytic hemoglobin, and remaining 1/3
Liver, spleen and bone marrow are laid in.Iron ion in serum is all combined with transferrinss, is the types of transportation of iron ion, referred to as
Serum levels of iron.Generally serum transferrin only have 30 % about combine with iron ion, still do not have 70% about not and iron ion knot
Close, the ability of this potential combination ferrum is referred to as unsaturated iron-binding capacity (uibc).In serum, transferrinss can be in conjunction with
Big iron is referred to as total iron binding capacity (tibc), and total iron binding capacity is equal to serum levels of iron and unsaturated iron-binding capacity sum, is appraiser
One of important indicator of body iron levels, clinically generally measures tibc and serum levels of iron and is used for evaluating body whether iron deficiency.
Complicated due to directly measuring tibc method, it is difficult to realize completely automatization, and uibc can more directly reflect storage in body
Deposit sideropenic situation, therefore begin with uibc in the world and measure the traditional total iron binding capacity of replacement.
Tibc measures and can be used for adjuvant clinical medical diagnosis on disease, and the such as disease such as iron deficiency anemia, acute hepatitises can cause tibc
Increase, liver cirrhosis, nephropathy, uremia, chronic infection, leukemia and heritability atransferrinemia etc. then can cause tibc to drop
Low.Tibc also can reflect the level of transferrinss indirectly, has important clinical meaning.
Realize full automation because total iron binding capacity is difficult to directly measure, therefore clinic is surveyed using transferrinss in recent years
Determine result to get more and more the report to estimate total iron binding capacity.Theoretically this method is more direct, and result is also more accurate,
But it is because that the mensure of serum transferrin is used mostly immunological method, expensive, complex operation, therefore with iron ion
Method based on mensure remains clinically the most frequently used.Reagent of the present invention, on the basis of ferene method, optimizes reactant
System, from amp buffer and add ascorbic acid, significantly improves the stability of reagent it was found that a kind of more stable insatiable hunger
With iron-binding capacity detectable.
Content of the invention
The present invention is intended to provide a kind of stable test kit for detecting unsaturated iron-binding capacity, the employing of this test kit
Ferene method., compared with conventional test kit, stability and linear dependence are better than conventional detection kit for this test kit,
Be conducive to reagent clinically popularization and application.
Ultimate principle:
In the alkaline buffer that excessive iron ion exists, the transferrinss not being combined with ferrum in serum are all tied with iron ion
Close, generate blue complex after remaining iron ion and reducing agent, developer effect, detect under 600nm wavelength.By calculating
In buffer, iron ion decrement just can calculate the unsaturated iron-binding capacity of serum.
The present invention is obtained through the following steps:
Reagent r1 consists of: the amp buffer of 100mmol/l ph 8.6,50mmol/l thiourea, 20mmol/l Ferrous ammonium sulfate,
200mmol/l oxammonium hydrochloride., 1% span80,10mmol/l sodium azide；Reagent r2 consists of: the sweet ammonia of 20mmol/l ph 3.2
Acid buffer, 2mmol/l ferene, 20mmol/l ascorbic acid, 1% span80,10mmol/l sodium azide.
Ratio when described reagent r1 and reagent r2 uses is r1:r2=4:1.
The test kit of the present invention is carried out on the automatic clinical chemistry analyzer with double reagent function, its concrete operation step
For:
Add normal saline, sample or calibration object 24 μ l, add the r1 reagent preincubate 5min of 240 μ l, add the r2 of 60 μ l
Reagent simultaneously reads absorbance a1, reads absorbance a2, and calculate δ a after reaction 5min.
Beneficial effects of the present invention:
1) reagent r1 adopts new buffer system, improves the stability of reagent；
2) reagent r2 adds ascorbic acid, enhances the stability of reagent, and will not produce impact to the accuracy of reagent；
3) accuracy of reagent and having good stability, low price, easy to use, be conducive to this reagent further in the market
Promote.
Brief description
A kind of contrast agents box correlation curve figure of accreditation on Fig. 1 test kit of the present invention and market；
A kind of contrast agents box stability curve figure of accreditation on Fig. 2 test kit of the present invention and market；
A kind of contrast agents box accuracy testing result of accreditation on Fig. 3 test kit of the present invention and market；
A kind of contrast agents box line related test results of accreditation on Fig. 4 test kit of the present invention and market；
A kind of contrast agents box Detection of Stability result of accreditation on Fig. 5 test kit of the present invention and market.
Specific embodiment
With reference to specific embodiment, the present invention is further described:
Embodiment 1
A kind of stable unsaturated iron-binding capacity detection kit, it includes reagent r1 and reagent r2.
Wherein reagent r1 consists of:
Ph 8.6 amp buffer 100mmol/l
Thiourea 50mmol/l
Ferrous ammonium sulfate 20mmol/l
Oxammonium hydrochloride. 200mmol/l
span80 1%
Sodium azide 10mmol/l
Reagent r2 consists of:
The glycine buffer 20mmol/l of ph 3.2
ferene 2mmol/l
Ascorbic acid 20mmol/l
span80 1%
Sodium azide 10mmol/l.
Reagent r1 described in the present embodiment and reagent r2, need to be initially charged buffer substance during configuration, after being transferred to suitable ph value, then add
Plus other composition.Test kit described in the present embodiment, when using, its assay method is using the Mai Rui with double reagent function
800 automatic clinical chemistry analyzers, are measured using performance rate method, and detection dominant wavelength is 600nm, and operation is as follows:
Add normal saline, sample or calibration object 24 μ l, add the r1 reagent preincubate 5min of 240 μ l, add the r2 of 60 μ l
Reagent simultaneously reads absorbance a1, reads absorbance a2, and calculate δ a after reaction 5min.δ a=a2- a1
Unsaturated iron-binding capacity content (μm ol/l)=(a measures ÷ a standard) × c standard.
Measure: detection sample absorbance change a standard: standard substance absorbance change
Embodiment 2
Accuracy validation is tested: using the unsaturated iron-binding capacity detectable of embodiment 1 as experimental group, market obtains accreditation
A kind of accuracy is good, good stability unsaturated iron-binding capacity detection kit is detected as a control group, 20 are faced
Bed serum sample is detected, testing result is as shown in Figure 3.Obtain the correlation curve (as shown in Figure 1) of two kinds of reagent, knot
Fruit shows, the correlation coefficient of two group reagent boxes is 0.9970, illustrates that both dependencys are relatively good.Prove that test kit of the present invention adds
And the component of change does not result in impact to its accuracy, test kit still keeps preferable accuracy.
Embodiment 3
Linear dependence checking test: prepare the high level sample that iron content is 80 μm of ol/l, be serially diluted with normal saline,
Prepare the sample of 6 variable concentrations, concentration be followed successively by 80 μm of ol/l, 64 μm of ol/l, 48 μm of ol/l, 32 μm of ol/l, 16 μm of ol/l,
0μmol/l.It is utilized respectively embodiment 1 reagent and comparison group reagent is detected, the sample of each concentration measures three times respectively, point
Do not average, testing result is as shown in Figure 4.
As shown in figure 4, embodiment 1 is all higher than 0.990 with compareing group reagent testing result correlation coefficient, and embodiment 1 is tried
The correlation coefficient of agent testing result is slightly larger than the correlation coefficient of comparison group reagent testing result, and this shows that reagent of the present invention has more
Good linear dependence.
Embodiment 4
Stability confirmatory experiment: store reagents in 2 DEG C～8 DEG C, the light protected environment of non-corrosiveness gas, detection embodiment 1 and
The stability of comparison group reagent.Same sample monthly chosen by two group reagents, measures and averages for three times, detection data such as Fig. 5 institute
Show.
Fig. 2 is obtained according to Fig. 5 detection data, Fig. 2 shows the lucifuge in 2 DEG C～8 DEG C, non-corrosiveness gas for embodiment 1 reagent
In environment, storage is stablized for 15 months, and compares group reagent and store 10 in 2 DEG C～8 DEG C, the light protected environment of non-corrosiveness gas
Start unstable after month, illustrate that reagent of the present invention is changed buffer and added reagent stability enhancing after Mannitol.
In sum, the unsaturated iron-binding capacity detection kit that the present invention provides, the buffer of reagent r1 is replaced by ph
Amp buffer for 8.6, will add ascorbic acid in reagent r2 simultaneously, improves the stability of test kit, linear dependence
Good, the accuracy of reagent is also preferable.Therefore, the unsaturated iron-binding capacity detection kit that the present invention provides is conducive in the market
Further promote the use of.
Claims (7)
1. a kind of stable unsaturated iron-binding capacity detection kit is it is characterised in that it comprises reagent r1 and reagent r2, wherein
Reagent r1 consists of: the amp buffer of 100mmol/l ph 8.6,50mmol/l thiourea, 20mmol/l Ferrous ammonium sulfate,
200mmol/l oxammonium hydrochloride., 1% span80,10mmol/l sodium azide；Reagent r2 consists of: the sweet ammonia of 20mmol/l ph 3.2
Acid buffer, 2mmol/l ferene, 20mmol/l ascorbic acid, 1% span80,10mmol/l sodium azide.
2. unsaturated iron-binding capacity detection kit according to claim 1 is it is characterised in that reagent r1 buffer is 25
DEG C, ph is 8.6 amp buffer (2-amino-2-methyl-1-propanol).
3. unsaturated iron-binding capacity detection kit according to claim 1 is it is characterised in that reagent r2 buffer is 25
DEG C, ph is 3.2 glycine buffer.
4. unsaturated iron-binding capacity detection kit according to claim 1 is it is characterised in that described developer is
Ferene(Panfuran Acetate disodium salt).
5. unsaturated iron-binding capacity detection kit according to claim 1 is it is characterised in that described surfactant is
Span80(sorbitan fatty acid ester).
6. unsaturated iron-binding capacity test kit according to claim 1 is it is characterised in that use automatic clinical chemistry analyzer
It is measured using end-point method, detection dominant wavelength is 600nm.
7. unsaturated iron-binding capacity test kit according to claim 1 is it is characterised in that described reagent r1 and reagent r2 makes
The ratio of used time is r1:r2=4:1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610742667.8A CN106370648A (en) | 2016-08-29 | 2016-08-29 | Detection kit with stable unsaturated iron-binding capacity |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610742667.8A CN106370648A (en) | 2016-08-29 | 2016-08-29 | Detection kit with stable unsaturated iron-binding capacity |
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| CN106370648A true CN106370648A (en) | 2017-02-01 |
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| CN201610742667.8A Pending CN106370648A (en) | 2016-08-29 | 2016-08-29 | Detection kit with stable unsaturated iron-binding capacity |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111007023A (en) * | 2019-12-11 | 2020-04-14 | 天津中成佳益生物科技有限公司 | Serum total iron binding force detection kit and preparation method and detection method thereof |
| CN111257549A (en) * | 2018-12-03 | 2020-06-09 | 深圳迈瑞生物医疗电子股份有限公司 | Kit and method for detecting unsaturated iron binding force in serum |
| CN115290584A (en) * | 2022-08-05 | 2022-11-04 | 中拓生物有限公司 | Stable unsaturated iron binding force determination kit |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102323430A (en) * | 2011-08-15 | 2012-01-18 | 北京利德曼生化股份有限公司 | Stable unsaturated iron bonding force determination kit |
| CN104483494A (en) * | 2014-12-22 | 2015-04-01 | 宁波美康生物科技股份有限公司 | Serum UIBC (unsaturated iron bonding capacity) detection kit |
-
2016
- 2016-08-29 CN CN201610742667.8A patent/CN106370648A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102323430A (en) * | 2011-08-15 | 2012-01-18 | 北京利德曼生化股份有限公司 | Stable unsaturated iron bonding force determination kit |
| CN104483494A (en) * | 2014-12-22 | 2015-04-01 | 宁波美康生物科技股份有限公司 | Serum UIBC (unsaturated iron bonding capacity) detection kit |
Non-Patent Citations (2)
| Title |
|---|
| 董振南等: "不饱和铁结合力试剂盒的方法学评价", 《中华检验医学杂志》 * |
| 蒋兴亮等: "双试剂直接分光光度法测定血清总铁结合力", 《国际检验医学杂志》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111257549A (en) * | 2018-12-03 | 2020-06-09 | 深圳迈瑞生物医疗电子股份有限公司 | Kit and method for detecting unsaturated iron binding force in serum |
| CN111007023A (en) * | 2019-12-11 | 2020-04-14 | 天津中成佳益生物科技有限公司 | Serum total iron binding force detection kit and preparation method and detection method thereof |
| CN111007023B (en) * | 2019-12-11 | 2024-05-10 | 天津中成佳益生物科技有限公司 | Serum total iron binding force detection kit, preparation method and detection method thereof |
| CN115290584A (en) * | 2022-08-05 | 2022-11-04 | 中拓生物有限公司 | Stable unsaturated iron binding force determination kit |
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Application publication date: 20170201 |