CN103901034A - Detection reagent strip for detecting microalbumin in urine and preparation method of detection reagent strip - Google Patents
Detection reagent strip for detecting microalbumin in urine and preparation method of detection reagent strip Download PDFInfo
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- CN103901034A CN103901034A CN201410178531.XA CN201410178531A CN103901034A CN 103901034 A CN103901034 A CN 103901034A CN 201410178531 A CN201410178531 A CN 201410178531A CN 103901034 A CN103901034 A CN 103901034A
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Abstract
The invention discloses a detection reagent strip for detecting microalbumin in urine. The detection reagent strip comprises a substrate and a filter paper piece, wherein the filter paper piece is attached on the substrate; the filter paper piece is sequentially immersed into an A solution and a B solution; the A solution is a buffered aqueous solution comprising sodium citrate, citric acid and concentrated hydrochloric acid; and the B solution is a methyl alcohol or ethanol solution comprising tetrahydrofuran, polypropylene glycol and sulfonephthalein dye. By utilizing the detection reagent strip prepared by the scheme, the sulfonephthalein dye can be quickly reacted with the microalbumin in the urine under acidic conditions, when the concentration of the microalbumin is in a range of 0-200mg/L, test paper presents obvious levels from colorlessness to mazarine, the levels are easy to visually distinguish and machine-readable, the measurement result is accurate and not disturbed by other ions in the urine, and the performance is stable. Meanwhile, a preparation method of the reagent strip is simple, convenient and rapid, the manufacturing cost is low, and the preparation method is suitable for large-scale popularization and application.
Description
Technical field
The present invention relates to urine detection field, particularly a kind of detection reagent strip that detects microalbumin in urine and preparation method thereof.
Background technology
Microalbumin refers to and in Human Urine, occurs micro-albumin.Albumin is the normal protein matter in a kind of blood, but in urine, only occurs minute quantity albumin under physiological condition.Under normal circumstances, the albumin in urine is few for body metabolism, answers < 20mg/L specific to every liter of urinary albumin.If the microalbumin after health check-up in discovery urine, within the scope of 20mg/L～200mg/L, just belongs to microalbuminuria, microalbuminuria reflection human kidney exceptional leakage protein.Conventionally application trace of albumin index is monitored the generation of ephrosis.The detection of trace of albumin is the most responsive, the most reliable diagnosis index of early detection ephrosis.By the numerical value of Urinary microalbumin, just can diagnose comparatively accurately the state of an illness in conjunction with incidence, symptom and medical history statement.
In prior art, detect microalbumin and conventionally adopt immunization, as immune-gold labeled, immunofluorescence etc., but the reagent strip complex production process of designing according to immunization, production cost is high, is difficult to large-scale promotion application.
Summary of the invention
For the deficiencies in the prior art part, the invention provides a kind of detection reagent strip that detects microalbumin in urine and preparation method thereof, compared to prior art, this preparation method is simple and efficient, and cost of manufacture is cheap, and reagent strip is easy to use, be convenient to preserve, testing result quick and precisely.
Technical scheme of the present invention is summarized as follows:
A detection reagent strip that detects microalbumin in urine, comprises substrate and filter paper, and described filter paper is attached on described substrate, in described filter paper, immerse and have A liquid and B liquid successively, wherein:
Described A liquid is to comprise: the aqueous buffer solution of 4.0wt%～4.5wt% sodium citrate, 4.2wt%～4.8wt% citric acid and 3.0wt%～3.5wt% concentrated hydrochloric acid;
Described B liquid is to comprise: methyl alcohol or the ethanolic solution of 4.0wt%～10wt% tetrahydrofuran, 0.5wt%～1.5wt% polypropylene glycol and 0.03wt%～0.15wt% sulfonephthalein dyestuff.
Preferably, the detection reagent strip of microalbumin in described detection urine, pH=2.0～3.5 of described A liquid.
Preferably, the detection reagent strip of microalbumin in described detection urine, described substrate material is Corvic, or polypropylene, or polymethylmethacrylate.
Preferably, the detection reagent strip of microalbumin in described detection urine, described filter paper is common quantitative filter paper or common qualitative filter paper.
Preferably, the detection reagent strip of microalbumin in described detection urine, described sulfonephthalein dyestuff is 5 ', 5 "-dinitro-3 ', 3 "-bis-iodo-3,4,5,6-tetrabromophenol sulfonphthalein.
A preparation method who detects the detection reagent strip of microalbumin in urine, it comprises the steps:
(1) configuration A liquid: 40～45g sodium citrate, 42～48g citric acid and 30～35mL concentrated hydrochloric acid are put into container, add water to 1000mL constant volume;
(2) configuration B liquid: 40～100mL tetrahydrofuran, 5～15mL polypropylene glycol and 0.3～1.5g sulfonephthalein dyestuff are put into container, add methyl alcohol or ethanol to 1000mL constant volume;
(3) filter paper is immersed in A liquid, after fully soaking, take out, the vacuum drying chamber that is placed in temperature and is 70 ℃ is dry, and be 10～20 minutes drying time;
(4) filter paper after drying is immersed in B liquid again, after fully soaking, take out, the vacuum drying chamber that is placed in temperature and is 60 ℃ is dry, and be 5～15 minutes drying time;
(5) dried filter paper and substrate are fixed, be cut into rectangle paper slip, make a kind of detection reagent strip that detects microalbumin in urine.
Preferably, the preparation method of the detection reagent strip of microalbumin in described detection urine, pH=2.0～3.5 of described A liquid.
Preferably, the preparation method of the detection reagent strip of microalbumin in described detection urine, described rectangle paper slip is of a size of 0.55 × 0.80cm.
The invention has the beneficial effects as follows: detection reagent strip prepared by this case, under acid condition, sulfonephthalein dyestuff can with urine in microalbumin react rapidly, in the time that microalbumin concentration is within the scope of 0～200mg/L, test paper presents from colourless and is easy to visual resolution and machine-readable to navy blue obvious color range, and measurement result is accurate, and be not subject to the interference of other ions in urine, stable performance; Meanwhile, the preparation method of this reagent strip is simple and efficient, and cost of manufacture is cheap, is applicable to large-scale promotion application.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail, to make those skilled in the art can implement according to this with reference to instructions word.
A preparation method who detects the detection reagent strip of microalbumin in urine, it comprises the steps:
(1) configuration A liquid: 40～45g sodium citrate, 42～48g citric acid and 30～35mL concentrated hydrochloric acid are put into container, add water to 1000mL constant volume, obtain the aqueous buffer solution of pH=2.0～3.5.
(2) configuration B liquid: 40～100mL tetrahydrofuran, 5～15mL polypropylene glycol (being preferably PPG2000) and 0.3～1.5g sulfonephthalein dyestuff are put into container, add methyl alcohol or ethanol to 1000mL constant volume; The purity of methyl alcohol or ethanol does not limit, and preferably absolute methanol or absolute ethyl alcohol can be also purity or lower (but as far as possible with the anhydrous system) of 80% (v/v%).Sulfonephthalein dyestuff is preferably 5 ', 5 "-dinitro-3 ', 3 "-bis-iodo-3,4,5,6-tetrabromophenol sulfonphthalein.
(3) filter paper is immersed in A liquid, after fully soaking, take out, the vacuum drying chamber that is placed in temperature and is 70 ℃ is dry, and be 10～20 minutes drying time.
(4) filter paper after drying is immersed in B liquid again, after fully soaking, take out, the vacuum drying chamber that is placed in temperature and is 60 ℃ is dry, and be 5～15 minutes drying time.
(5) dried filter paper and substrate are fixed, be cut into the rectangle paper slip that is of a size of 0.55 × 0.80cm, make a kind of detection reagent strip that detects microalbumin in urine.This substrate material can be Corvic, or polypropylene, or polymethylmethacrylate; And filter paper can be common quantitative filter paper or common qualitative filter paper.
Embodiment 1
The present embodiment provides a kind of preparation method of the detection reagent strip that detects microalbumin in urine, and it comprises the steps:
(1) configuration A liquid: 40g sodium citrate, 42g citric acid and 30mL concentrated hydrochloric acid are put into container, add water to 1000mL constant volume;
(2) configuration B liquid: 40mL tetrahydrofuran, 5mL polypropylene glycol and 0.3g sulfonephthalein dyestuff are put into container, add methyl alcohol to 1000mL constant volume;
(3) filter paper is immersed in A liquid, after fully soaking, take out, the vacuum drying chamber that is placed in temperature and is 70 ℃ is dry, and be 10 minutes drying time;
(4) filter paper after drying is immersed in B liquid again, after fully soaking, take out, the vacuum drying chamber that is placed in temperature and is 60 ℃ is dry, and be 5 minutes drying time;
(5) dried filter paper and substrate are fixed, be cut into rectangle paper slip, make a kind of detection reagent strip that detects microalbumin in urine.
Embodiment 2
The present embodiment provides a kind of preparation method of the detection reagent strip that detects microalbumin in urine, and it comprises the steps:
(1) configuration A liquid: 45g sodium citrate, 48g citric acid and 35mL concentrated hydrochloric acid are put into container, add water to 1000mL constant volume;
(2) configuration B liquid: 100mL tetrahydrofuran, 15mL polypropylene glycol and 1.5g sulfonephthalein dyestuff are put into container, add ethanol to 1000mL constant volume;
(3) filter paper is immersed in A liquid, after fully soaking, take out, the vacuum drying chamber that is placed in temperature and is 70 ℃ is dry, and be 20 minutes drying time;
(4) filter paper after drying is immersed in B liquid again, after fully soaking, take out, the vacuum drying chamber that is placed in temperature and is 60 ℃ is dry, and be 15 minutes drying time;
(5) dried filter paper and substrate are fixed, be cut into rectangle paper slip, make a kind of detection reagent strip that detects microalbumin in urine.
Embodiment 3
The present embodiment provides a kind of preparation method of the detection reagent strip that detects microalbumin in urine, and it comprises the steps:
(1) configuration A liquid: 42.5g sodium citrate, 45g citric acid and 32.5mL concentrated hydrochloric acid are put into container, add water to 1000mL constant volume;
(2) configuration B liquid: 70mL tetrahydrofuran, 10mL polypropylene glycol and 0.9g sulfonephthalein dyestuff are put into container, add methyl alcohol to 1000mL constant volume;
(3) filter paper is immersed in A liquid, after fully soaking, take out, the vacuum drying chamber that is placed in temperature and is 70 ℃ is dry, and be 15 minutes drying time;
(4) filter paper after drying is immersed in B liquid again, after fully soaking, take out, the vacuum drying chamber that is placed in temperature and is 60 ℃ is dry, and be 10 minutes drying time;
(5) dried filter paper and substrate are fixed, be cut into rectangle paper slip, make a kind of detection reagent strip that detects microalbumin in urine.
Embodiment 4
The present embodiment provides a kind of preparation method of the detection reagent strip that detects microalbumin in urine, and it comprises the steps:
(1) configuration A liquid: 41.7g sodium citrate, 45.1g citric acid and 32mL concentrated hydrochloric acid are put into container, add water to 1000mL constant volume;
(2) configuration B liquid: by 50mL tetrahydrofuran, 10mL polypropylene glycol and 0.58g 5 ', 5 "-dinitro-3 ', 3 "-bis-iodo-3,4,5,6-tetrabromophenol sulfonphthalein is put into container, adds ethanol to 1000mL constant volume;
(3) filter paper is immersed in A liquid, after fully soaking, take out, the vacuum drying chamber that is placed in temperature and is 70 ℃ is dry, and be 15 minutes drying time;
(4) filter paper after drying is immersed in B liquid again, after fully soaking, take out, the vacuum drying chamber that is placed in temperature and is 60 ℃ is dry, and be 15 minutes drying time;
(5) dried filter paper and substrate are fixed, be cut into rectangle paper slip, make a kind of detection reagent strip that detects microalbumin in urine.
Embodiment 5
The preparation method of standard color comparison card:
(1) the bovine serum albumin(BSA) standard solution of configuration 0mg/L, 20mg/L, 80mg/L, 150mg/L;
(2) detection reagent strip above-described embodiment 1～4 being prepared is dipped in respectively in four kinds of above-mentioned bovine serum albumin(BSA) standard solution, after 2 seconds, takes out, and dries, and after 1 minute, colour developing completely;
(3) prepare colorimetric card according to the colour developing situation of each test paper, the color range of colorimetric card is that white is to navy blue four color ranges.
Embodiment 6
Microalbumin detects the using method of reagent strip:
The detection reagent strip that embodiment 1～4 is prepared immerses in liquid to be measured, after 2 seconds, take out, dry, after 1 minute, colour developing completely, compare with standard color comparison card subsequently, with the concentration that detects the corresponding concentration of color on the immediate standard color comparison card of color on reagent strip and be liquid to be measured, if between two color ranges, can estimate and read microalbumin concentration.
Embodiment 7
Urinary microalbumin detects the using method of reagent strip on instrument:
(1) the bovine serum albumin(BSA) standard solution of configuration 0mg/L, 20mg/L, 80mg/L, 150mg/L, 200mg/L;
(2) detection reagent strip embodiment 1～4 being prepared is dipped in respectively in five kinds of above-mentioned bovine serum albumin(BSA) standard solution;
(3) use Urine Analyzer to measure the reflectivity of 1 minute, according to 5 point-rendering typical curves;
(4) while use, the microdose urine protein detection reagent strip preparing according to embodiment is immersed in liquid to be measured and taken out, after 1 minute, use Urine Analyzer to measure reflectivity, the reflectivity recording is liquid microalbumin concentration to be measured with concentration corresponding on typical curve.
Although embodiment of the present invention are open as above, but it is not restricted to listed utilization in instructions and embodiment, it can be applied to various applicable the field of the invention completely, for those skilled in the art, can easily realize other modification, therefore do not deviating under the universal that claim and equivalency range limit, the present invention is not limited to specific details.
Claims (8)
1. a detection reagent strip that detects microalbumin in urine, comprises substrate and filter paper, and described filter paper is attached on described substrate, it is characterized in that, in described filter paper, immerse and have A liquid and B liquid successively, wherein:
Described A liquid is to comprise: the aqueous buffer solution of 4.0wt%～4.5wt% sodium citrate, 4.2wt%～4.8wt% citric acid and 3.0wt%～3.5wt% concentrated hydrochloric acid;
Described B liquid is to comprise: methyl alcohol or the ethanolic solution of 4.0wt%～10wt% tetrahydrofuran, 0.5wt%～1.5wt% polypropylene glycol and 0.03wt%～0.15wt% sulfonephthalein dyestuff.
2. the detection reagent strip of microalbumin in detection urine according to claim 1, is characterized in that pH=2.0～3.5 of described A liquid.
3. the detection reagent strip of microalbumin in detection urine according to claim 1, is characterized in that, described substrate material is Corvic, or polypropylene, or polymethylmethacrylate.
4. the detection reagent strip of microalbumin in detection urine according to claim 1, is characterized in that, described filter paper is common quantitative filter paper or common qualitative filter paper.
5. the detection reagent strip of microalbumin in detection urine according to claim 1, is characterized in that, described sulfonephthalein dyestuff is 5 ', 5 "-dinitro-3 ', 3 "-bis-iodo-3,4,5,6-tetrabromophenol sulfonphthalein.
6. a preparation method who detects the detection reagent strip of microalbumin in urine, its feature comprises the steps:
(1) configuration A liquid: 40～45g sodium citrate, 42～48g citric acid and 30～35mL concentrated hydrochloric acid are put into container, add water to 1000mL constant volume;
(2) configuration B liquid: 40～100mL tetrahydrofuran, 5～15mL polypropylene glycol and 0.3～1.5g sulfonephthalein dyestuff are put into container, add methyl alcohol or ethanol to 1000mL constant volume;
(3) filter paper is immersed in A liquid, after fully soaking, take out, the vacuum drying chamber that is placed in temperature and is 70 ℃ is dry, and be 10～20 minutes drying time;
(4) filter paper after drying is immersed in B liquid again, after fully soaking, take out, the vacuum drying chamber that is placed in temperature and is 60 ℃ is dry, and be 5～15 minutes drying time;
(5) dried filter paper and substrate are fixed, be cut into rectangle paper slip, make a kind of detection reagent strip that detects microalbumin in urine.
7. the preparation method of the detection reagent strip of microalbumin in detection urine according to claim 6, is characterized in that pH=2.0～3.5 of described A liquid.
8. the preparation method of the detection reagent strip of microalbumin in detection urine according to claim 6, is characterized in that, described rectangle paper slip is of a size of 0.55 × 0.80cm.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105388146A (en) * | 2015-10-20 | 2016-03-09 | 北京中生金域诊断技术股份有限公司 | Kit for simultaneously detecting sodium, creatinine and microalbumin in urine |
| CN105651765A (en) * | 2015-12-28 | 2016-06-08 | 东北农业大学 | Rapid protein test paper, and production method and detection method thereof |
| CN106596913A (en) * | 2017-01-18 | 2017-04-26 | 重庆普西医疗设备有限公司 | Preparation method of quantitative analysis test paper for urine |
| CN106645676A (en) * | 2017-01-18 | 2017-05-10 | 重庆普西医疗设备有限公司 | Multi-term urine analysis test paper and preparation method thereof |
| CN108120714A (en) * | 2017-12-20 | 2018-06-05 | 吉林省汇酉生物技术股份有限公司 | A kind of reagent for detecting urine microdose urine protein and its application |
| CN108226147A (en) * | 2017-12-27 | 2018-06-29 | 李宏奎 | A kind of microdose urine protein detection reagent item and preparation method thereof |
| CN113376150A (en) * | 2021-06-10 | 2021-09-10 | 吉林基蛋生物科技有限公司 | Urine microalbumin dry chemical detection test paper and preparation method thereof |
| CN114034693A (en) * | 2021-11-04 | 2022-02-11 | 广州达安基因股份有限公司 | Combined test paper for detecting urine creatinine urine microalbumin semiquantitative, preparation method and application thereof |
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| RU2692062C1 (en) * | 2018-06-19 | 2019-06-20 | Евгений Александрович Ширшин | Method of producing a color pattern and a method of analyzing colorimetric test strips using said pattern |
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| CN105388146A (en) * | 2015-10-20 | 2016-03-09 | 北京中生金域诊断技术股份有限公司 | Kit for simultaneously detecting sodium, creatinine and microalbumin in urine |
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| CN108226147A (en) * | 2017-12-27 | 2018-06-29 | 李宏奎 | A kind of microdose urine protein detection reagent item and preparation method thereof |
| CN108226147B (en) * | 2017-12-27 | 2021-06-04 | 李宏奎 | Urine microalbumin detection reagent strip and preparation method thereof |
| CN113376150A (en) * | 2021-06-10 | 2021-09-10 | 吉林基蛋生物科技有限公司 | Urine microalbumin dry chemical detection test paper and preparation method thereof |
| CN114034693A (en) * | 2021-11-04 | 2022-02-11 | 广州达安基因股份有限公司 | Combined test paper for detecting urine creatinine urine microalbumin semiquantitative, preparation method and application thereof |
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