CN106770251B - Urine analysis test paper and preparation method thereof - Google Patents
Urine analysis test paper and preparation method thereof Download PDFInfo
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- CN106770251B CN106770251B CN201611252827.7A CN201611252827A CN106770251B CN 106770251 B CN106770251 B CN 106770251B CN 201611252827 A CN201611252827 A CN 201611252827A CN 106770251 B CN106770251 B CN 106770251B
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Abstract
The invention relates to a urine analysis test paper and a preparation method thereof, wherein the test paper comprises a plurality of detection blocks and a plurality of isolation blocks, a liquid absorbing area is made of a water absorbing material, the isolation blocks are made of a hydrophobic material, the detection blocks are provided with chemical reagents for detecting various items, a substrate is arranged below the test paper, and a through sample injection hole is formed in the center of the substrate. According to the urine analysis test paper and the preparation method thereof, urine is dripped into each detection block through the hydrophobic barrier interval by each detection block through the through holes, so that the urine reacts with the reagent on each detection block simultaneously, a plurality of test modules are integrated on the same miniature test paper, the amount of the used urine is small, the whole detection process can be finished instantly, the urine does not need to be dripped carefully in sequence like a conventional urine test paper, and the detection cost can be greatly reduced while the detection efficiency is improved.
Description
Technical Field
The invention relates to the field of in-vitro diagnosis, in particular to urine analysis test paper and a preparation method thereof.
Background
The dry chemical analysis technology refers to that a liquid detection sample is directly added to a commercial dry reagent strip specially produced for different projects, water of the detected sample is taken as a solvent to cause specific chemical reaction, so that chemical analysis is performed, and the dry chemical analysis technology is an analysis method based on an enzyme method, is widely applied to urine detection and analysis, and determines whether urine contains a certain component or not by changing the color of a special test paper through the chemical reaction of various special test papers and corresponding components in the urine. The reagent of multiple detection items can be integrated on the existing test strip, so that one test strip comprises a plurality of item detection blocks, but the existing test strip is large in size and generally in a narrow strip shape, urine to be detected needs to be dripped in the block of a specific detection item in a divided and accurate manner, the required urine sample amount is large, and the detection efficiency is low.
Disclosure of Invention
The invention aims to solve the defects of the prior art and provides urine analysis test paper and a preparation method thereof.
The invention adopts the following technical scheme to realize the aim: the urine analysis test paper is characterized by comprising a test paper and a substrate, wherein the test paper comprises a plurality of liquid absorbing areas and detection areas which are in rectangular bar shapes, the liquid absorbing areas and the detection areas are arranged at intervals, each detection area consists of a plurality of detection blocks and a plurality of isolation blocks, the isolation blocks and the detection blocks are arranged at intervals,
the liquid absorbing area is made of water absorbing material, the isolating block is made of hydrophobic material,
the substrate is arranged below the test paper, and a through sample injection hole is formed in the center of the substrate.
In particular, three sample injection holes are arranged, and the three sample injection holes respectively correspond to different liquid absorption areas.
Particularly, the baffle is arranged upwards at the peripheral edge of the substrate, and the test paper is fixed on the substrate by the baffle.
Particularly, the water absorbing material for preparing the liquid absorbing area is one of water absorbing paper, glass fiber membrane and polyester fiber membrane; the hydrophobic material of the spacer is one of a curable polymer and a photoresist.
The invention also provides a preparation method of the urine analysis test paper, which is characterized by comprising the following steps:
(1) cutting a water-absorbing material into 10mm multiplied by 7mm in size to serve as a test paper base material, dipping the test paper base material with a hydrophobic material drop, polymerizing and solidifying the test paper base material to form a separation block serving as a hydrophobic barrier, and separating the test paper base material into a plurality of detection blocks, wherein the periphery of each detection block is surrounded by the hydrophobic barrier;
(2) respectively performing chemical reagent dripping and treatment on the plurality of detection blocks, and respectively preparing the plurality of detection blocks into project detection blocks for detecting different projects;
the project detection block is selected from a glucose detection block, a PH value detection block, a protein detection block, a microalbumin detection block, a specific gravity detection block, a ketone detection block, a bilirubin detection block, a urobilinogen detection block, a nitrite detection block, a leucocyte detection block, an ascorbic acid detection block, a occult blood detection block, a uric acid detection block, a creatinine detection block and a calcium detection block, and the preparation method of each project detection block specifically comprises the following steps:
preparation of glucose test block
Weighing 0.19g of citric acid, adding into 10ml of ultrapure water, stirring thoroughly until the citric acid is completely dissolved, regulating the pH of the solution to 6.0 by using 0.1mol/L sodium hydroxide, adding 0.025g of tetramethylbenzidine, 920u of glucose oxidase and 19u of peroxidase into the buffer solution, and stirring until the citric acid is completely dissolved; using a pipette or an automatic sample application instrument to sample the prepared solution to a glucose detection area, wherein the sample size is 0.5-1 μl, and drying at 25-35deg.C after sample application;
preparation of PH value detection block
1.2g of methyl red, 0.8g of bromothymol blue and 0.8ml of ethanol are weighed and added into 10ml of ultrapure water, and the mixture is fully stirred until the mixture is completely dissolved; using a pipettor or an automatic sample application instrument to sample the prepared solution in the pH value detection area, wherein the sample size is 0.5-1 μl, and drying at 50-80deg.C after sample application is completed;
protein detection Block preparation
Weighing 0.095g of citric acid, adding into 10ml of ultrapure water, fully stirring until the citric acid is completely dissolved, adjusting the pH of the solution to 2.0-3.0 by using 0.1mol/L of sodium hydroxide and 0.1mol/L of hydrochloric acid, adding 0.8ml of ethanol, 0.025g of polyvinyl alcohol and 0.033g of tetrabromophenol blue into the buffer solution, and stirring until the solution is completely dissolved; using a pipettor or an automatic sample application instrument to sample the prepared solution to a protein detection area, wherein the sample size is 0.5-1 μl, and drying at 50-80deg.C after sample application is completed;
preparation of microalbumin detection block
1.9g of citric acid is weighed and added into 10ml of ultrapure water, fully stirred until the citric acid is completely dissolved, the PH of the solution is regulated to 2.0 to 3.0 by using 1mol/L of sodium hydroxide and 1mol/L of hydrochloric acid, 0.01g of sulfophthalein dye is added into the buffer solution, and the solution is stirred until the solution is completely dissolved; using a pipette or an automatic sample application instrument to sample the prepared solution to a trace albumin detection area, wherein the sample size is 0.5-1 μl, and drying at 50-70deg.C after sample application;
specific gravity detection block preparation
Weighing 0.012g of trimetaphosphate, 0.018g of bromothymol blue and 0.05g of tetramethylammonium hydroxide, adding into 10ml of ultrapure water, fully stirring until the solution is completely dissolved, using a pipette or an automatic sample application instrument to sample the prepared solution in a specific gravity detection area, wherein the sample size is 0.5-1 mu l, and drying at 50-80 ℃ after the sample application is finished;
preparation of ketone body detection block
Weighing 0.5g of sodium hydroxide, adding 0.125g of sodium nitrosoferricyanide into 10ml of ultrapure water, fully stirring until the sodium hydroxide is completely dissolved, using a pipette or an automatic sample application instrument to sample the prepared solution in a ketone detection area, wherein the sample size is 0.5-1 μl, and drying at 35-50 ℃ after sample application is completed;
bilirubin detection block preparation
Weighing 0.95g of citric acid, adding into 10ml of ultrapure water, fully stirring until the citric acid is completely dissolved, adjusting the pH of the solution to 5.0 by using 1mol/L sodium hydroxide, adding 0.35g of 2,6 dichloro diazonium salt fluoborate into the buffer solution, and stirring until the boric acid is completely dissolved; using a pipettor or an automatic spotting instrument to sample the prepared solution in the bilirubin detection area, wherein the sample size is 0.5-1 μl, and drying at 50-70deg.C after the sample application is completed;
preparation of urine bilinogen detection block
Weighing 0.95g of citric acid, adding into 10ml of ultrapure water, fully stirring until the citric acid is completely dissolved, adjusting the pH of the solution to 5.0 by using 1mol/L sodium hydroxide, adding 0.35g of p-xylene diazonium boron tetrafluoride into the buffer solution, and stirring until the boron tetrafluoride is completely dissolved; using a pipettor or an automatic sample application instrument to sample the prepared solution to the urine bilinogen detection area, wherein the sample size is 0.5-1 μl, and drying at 50-70deg.C after sample application;
nitrite detection block preparation
0.08g of p-aminobenzene arsinic acid, 0.09g of N- (1-naphthylamine) -ethylenediamine and 0.01g of sodium periodate are weighed and added into 10ml of ultrapure water, and the mixture is fully stirred until the sodium periodate is completely dissolved; using a pipettor or an automatic sample application instrument to sample the prepared solution to a nitrite detection area, wherein the sample size is 0.5-1 μl, and drying at 30-40deg.C after sample application;
preparation of leucocyte detection blocks
Weighing 0.25g of disodium hydrogen phosphate, 0.03g of sodium dihydrogen phosphate and 0.09g of sodium chloride, adding into 10ml of ultrapure water, fully stirring until the disodium hydrogen phosphate, the sodium chloride and the sodium chloride are completely dissolved, adjusting the pH of a solution to 9.0 by using 0.1mol/L sodium hydroxide, adding 0.04g of pyrrole ester, 0.02g of diazonium salt and 0.075g of imidazole into the buffer solution, and stirring until the sodium hydrogen phosphate, the sodium dihydrogen phosphate and the sodium chloride are completely dissolved; using a pipette or an automatic spotting instrument to spot the prepared solution on a leukocyte detection area, wherein the sample size is 0.5-1 μl, and drying at 50-70deg.C after the spot is finished;
preparation of ascorbic acid detection block
Weighing 0.08g of 2, 6-dichlorophenol indophenol, adding into 10ml of ultrapure water, and fully stirring until the indophenol is completely dissolved; using a pipette or an automatic spotting instrument to spot the prepared solution on an ascorbic acid detection area, wherein the sample size is 0.5-1 μl, and drying at 40-60deg.C after the spot is finished;
preparation of occult blood detection block
Weighing 0.17g of disodium hydrogen phosphate, 0.03g of sodium dihydrogen phosphate and 0.09g of sodium chloride, adding into 10ml of ultrapure water, fully stirring until the disodium hydrogen phosphate, the sodium chloride and the sodium chloride are completely dissolved, adjusting the pH of a solution to 6.5 by using 0.1mol/L of sodium hydroxide and 0.1mol/L of hydrochloric acid, taking 0.012g of carbamide peroxide, 0.08g of o-tolidine, 0.05g of 2-aminothiazole acetate and 2.6g of potassium iodate, adding into the buffer solution, and stirring until the potassium hydrogen phosphate, the sodium hydrogen phosphate and the sodium chloride are completely dissolved; using a pipettor or an automatic sample application instrument to sample the prepared solution to the occult blood detection area, wherein the sample size is 0.5-1 μl, and drying at 30-45deg.C after sample application;
uric acid detection block preparation
Weighing 0.20g of disodium hydrogen phosphate, 0.03g of sodium dihydrogen phosphate and 0.09g of sodium chloride, adding into 10ml of ultrapure water, fully stirring until the disodium hydrogen phosphate, the sodium chloride and the sodium chloride are completely dissolved, adjusting the pH of a solution to 7.2 by using 0.1mol/L of sodium hydroxide and 0.1mol/L of hydrochloric acid, adding 0.06g of 4-aminoantipyrine, 0.04g of 2-hydroxy-3, 5-dichlorobenzenesulfonic acid, 15u of uricase and 25u of peroxidase into the buffer solution, and stirring until the solution is completely dissolved; using a pipettor or an automatic sample application instrument to sample the prepared solution to the uric acid detection area, wherein the sample size is 0.5-1 μl, and drying at 25-35deg.C after sample application;
preparation of creatinine detection block
Weighing 0.12G boric acid, adding into 10ml ultrapure water, fully stirring until the boric acid is completely dissolved, regulating the pH of the solution to 12 by using 0.1mol/L sodium hydroxide and 0.1mol/L hydrochloric acid, adding 0.05G potassium chloride, 0.02G polyvinylpyrrolidone, 0.001G orange G and 0.005G 3.5-dinitrobenzoic acid into the buffer solution, and stirring until the boric acid is completely dissolved; using a pipettor or an automatic sample application instrument to sample the prepared solution in the creatinine detection area, wherein the sample size is 0.5-1 μl, and drying at 40-60deg.C after the sample application is completed;
preparation of calcium detection block
Weighing 0.19g of lemon yellow, adding into 10ml of ultrapure water, fully stirring until the lemon yellow is completely dissolved, regulating the pH of a solution to 9.5 by using 0.1mol/L sodium hydroxide and 0.1mol/L hydrochloric acid, taking 0.035g of o-cresolphthalein complexing ketone, adding 0.1ml of Triton x-100 and 0.035g of EGTA into the buffer solution, and stirring until the lemon yellow is completely dissolved; using a pipettor or an automatic sample application instrument to sample the prepared solution in the creatinine detection area, wherein the sample size is 0.5-1 μl, and drying at 60-80deg.C after sample application is completed;
(3) the prepared test paper is fixed in a base plate with three holes at the bottom.
The beneficial effects of the invention are as follows: the invention provides a urine analysis test paper capable of simultaneously detecting a plurality of items and a preparation method thereof, wherein each detection block is separated by a hydrophobic barrier, urine is dripped into each detection block through a through hole, so that the urine and reagents on each detection block react simultaneously, and a plurality of test modules are integrated on the same miniature test paper, so that the amount of the urine used is small, the whole detection process can be finished instantaneously, the urine does not need to be dripped in sequence like a conventional urine test paper, the detection efficiency is improved, and the detection cost can be greatly reduced.
Drawings
FIG. 1 is a schematic diagram of the structure of the present invention;
FIG. 2 is a schematic view of the structure of the bottom part of the present invention;
in the figure: 1-a liquid absorption zone; 2-a detection block; 3-isolating blocks; 4-a substrate; 5-baffle plates; 6-sample injection holes;
the embodiments of the present invention will be described in detail below with reference to the accompanying drawings.
Detailed Description
The invention is further illustrated by the following examples in conjunction with the accompanying drawings:
example 1
As shown in fig. 1 and 2, the urine analysis test paper comprises a test paper and a substrate 4, wherein the test paper comprises three liquid absorbing areas 1 with rectangular strip shapes and four detection areas with rectangular strip shapes, the four liquid absorbing areas 1 are arranged with the three detection areas at intervals, each detection area consists of four detection blocks 2 and five isolation blocks 3, the five isolation blocks 3 and the four detection blocks 2 are arranged at intervals,
the liquid absorbing area 1 is made of a polyester fiber film, the spacer 3 is made of a photoresist,
the base plate 4 is arranged below the test paper, three through sample injection holes 6 are arranged in the center of the base plate 4, the three sample injection holes 6 respectively correspond to different liquid absorption areas 1,
baffle plates 5 are arranged upwards at the peripheral edges of the substrate 4, and the test paper is fixed on the substrate 4 by the baffle plates 5.
A preparation method of urine analysis test paper comprises the following steps:
(1) cutting a water-absorbing material into 10mm multiplied by 7mm in size to serve as a test paper base material, dripping a curable polymer onto the test paper base material, polymerizing and curing the curable polymer to form a separation block 3 serving as a hydrophobic barrier, and separating the test paper base material into three rows of four detection blocks 2 each, wherein the periphery of each detection block 2 is surrounded by the hydrophobic barrier;
(2) performing chemical reagent dripping and treatment on twelve detection blocks 2 respectively, and preparing the twelve detection blocks 2 into project detection blocks for detecting different projects respectively;
the project detection block is a glucose detection block, a pH value detection block, a protein detection block, a microalbumin detection block, a specific gravity detection block, a ketone detection block, a leucocyte detection block, an ascorbic acid detection block, a occult blood detection block, a uric acid detection block, a creatinine detection block and a calcium detection block, and the preparation method of each project detection block specifically comprises the following steps:
preparation of glucose test block
Weighing 0.19g of citric acid, adding into 10ml of ultrapure water, stirring thoroughly until the citric acid is completely dissolved, regulating the pH of the solution to 6.0 by using 0.1mol/L sodium hydroxide, adding 0.025g of tetramethylbenzidine, 920u of glucose oxidase and 19u of peroxidase into the buffer solution, and stirring until the citric acid is completely dissolved; spotting the prepared solution on a glucose detection area by using a pipette, wherein the sample size is 0.5 mu l, and drying at 25 ℃ after spotting is finished;
preparation of PH value detection block
1.2g of methyl red, 0.8g of bromothymol blue and 0.8ml of ethanol are weighed and added into 10ml of ultrapure water, and the mixture is fully stirred until the mixture is completely dissolved; spotting the prepared solution on the pH value detection area by using a pipette, wherein the sample size is 0.5 mu l, and drying at 50 ℃ after spotting is finished;
protein detection Block preparation
Weighing 0.095g of citric acid, adding into 10ml of ultrapure water, fully stirring until the citric acid is completely dissolved, regulating the pH of the solution to 2.0 by using 0.1mol/L of sodium hydroxide and 0.1mol/L of hydrochloric acid, adding 0.8ml of ethanol, 0.025g of polyvinyl alcohol and 0.033g of tetrabromophenol blue into the buffer solution, and stirring until the tetrabromophenol blue is completely dissolved; spotting the prepared solution on a protein detection area by using a pipette, wherein the sample size is 0.5 mu l, and drying at 50 ℃ after spotting is finished;
preparation of microalbumin detection block
1.9g of citric acid is weighed and added into 10ml of ultrapure water, the mixture is fully stirred until the mixture is completely dissolved, 1mol/L of sodium hydroxide and 1mol/L of hydrochloric acid are used for regulating the pH of the solution to 2.0, 0.01g of sulfophthalein dye is added into the buffer solution, and the mixture is stirred until the mixture is completely dissolved; spotting the prepared solution on a trace albumin detection area by using a pipette, wherein the sample size is 0.5 mu l, and drying at 50 ℃ after spotting is finished;
specific gravity detection block preparation
Weighing 0.012g of trimetaphosphate, 0.018g of bromothymol blue and 0.05g of tetramethylammonium hydroxide, adding into 10ml of ultrapure water, fully stirring until the solution is completely dissolved, using a pipette to sample a specific gravity detection area, sampling the solution with a sample size of 0.5 mu l, and drying at 50 ℃ after sample application;
preparation of ketone body detection block
Weighing 0.5g of sodium hydroxide, adding 0.125g of sodium nitrosoferricyanide into 10ml of ultrapure water, fully stirring until the sodium hydroxide is completely dissolved, using a pipettor to sample the ketone detection area with the prepared solution, and drying at 35 ℃ after sample application is finished, wherein the sample size is 0.5 μl;
preparation of leucocyte detection blocks
Weighing 0.25g of disodium hydrogen phosphate, 0.03g of sodium dihydrogen phosphate and 0.09g of sodium chloride, adding into 10ml of ultrapure water, fully stirring until the disodium hydrogen phosphate, the sodium chloride and the sodium chloride are completely dissolved, adjusting the pH of a solution to 9.0 by using 0.1mol/L sodium hydroxide, adding 0.04g of pyrrole ester, 0.02g of diazonium salt and 0.075g of imidazole into the buffer solution, and stirring until the sodium hydrogen phosphate, the sodium dihydrogen phosphate and the sodium chloride are completely dissolved; spotting the prepared solution on a leukocyte detection area by using a pipette, wherein the sample size is 0.5 mu l, and drying at 50 ℃ after spotting is finished;
preparation of ascorbic acid detection block
Weighing 0.08g of 2, 6-dichlorophenol indophenol, adding into 10ml of ultrapure water, and fully stirring until the indophenol is completely dissolved; spotting the prepared solution on an ascorbic acid detection area by using a pipette, wherein the sample size is 0.5 mu l, and drying at 40 ℃ after spotting is finished;
preparation of occult blood detection block
Weighing 0.17g of disodium hydrogen phosphate, 0.03g of sodium dihydrogen phosphate and 0.09g of sodium chloride, adding into 10ml of ultrapure water, fully stirring until the disodium hydrogen phosphate, the sodium chloride and the sodium chloride are completely dissolved, adjusting the pH of a solution to 6.5 by using 0.1mol/L of sodium hydroxide and 0.1mol/L of hydrochloric acid, taking 0.012g of carbamide peroxide, 0.08g of o-tolidine, 0.05g of 2-aminothiazole acetate and 2.6g of potassium iodate, adding into the buffer solution, and stirring until the potassium hydrogen phosphate, the sodium hydrogen phosphate and the sodium chloride are completely dissolved; spotting the prepared solution on a occult blood detection area by using a pipette, wherein the sample size is 0.5 mu l, and drying at 30 ℃ after the spotting is finished;
uric acid detection block preparation
Weighing 0.20g of disodium hydrogen phosphate, 0.03g of sodium dihydrogen phosphate and 0.09g of sodium chloride, adding into 10ml of ultrapure water, fully stirring until the disodium hydrogen phosphate, the sodium chloride and the sodium chloride are completely dissolved, adjusting the pH of a solution to 7.2 by using 0.1mol/L of sodium hydroxide and 0.1mol/L of hydrochloric acid, adding 0.06g of 4-aminoantipyrine, 0.04g of 2-hydroxy-3, 5-dichlorobenzenesulfonic acid, 15u of uricase and 25u of peroxidase into the buffer solution, and stirring until the solution is completely dissolved; using an automatic sample application instrument to sample the prepared solution to the uric acid detection area, wherein the sample size is 1 μl, and drying at 35 ℃ after sample application is completed;
preparation of creatinine detection block
Weighing 0.12G boric acid, adding into 10ml ultrapure water, fully stirring until the boric acid is completely dissolved, regulating the pH of the solution to 12 by using 0.1mol/L sodium hydroxide and 0.1mol/L hydrochloric acid, adding 0.05G potassium chloride, 0.02G polyvinylpyrrolidone, 0.001G orange G and 0.005G 3.5-dinitrobenzoic acid into the buffer solution, and stirring until the boric acid is completely dissolved; using an automatic sample application instrument to sample the prepared solution to the creatinine detection region, wherein the sample size is 1 μl, and drying at 60deg.C after sample application is completed;
preparation of calcium detection block
Weighing 0.19g of lemon yellow, adding into 10ml of ultrapure water, fully stirring until the lemon yellow is completely dissolved, regulating the pH of a solution to 9.5 by using 0.1mol/L sodium hydroxide and 0.1mol/L hydrochloric acid, taking 0.035g of o-cresolphthalein complexing ketone, adding 0.1ml of Triton x-100 and 0.035g of EGTA into the buffer solution, and stirring until the lemon yellow is completely dissolved; using an automatic sample application instrument to sample the prepared solution to the creatinine detection region, wherein the sample size is 1 μl, and drying at 80deg.C after sample application is completed;
(3) the prepared test paper is fixed in a base plate 4 with three sample injection holes 6 at the bottom.
Example 2
The urine analysis test paper comprises test paper and a substrate 4, wherein the test paper comprises three liquid absorbing areas 1 with rectangular bar shapes and four detection areas with rectangular bar shapes, the four liquid absorbing areas 1 are arranged with the three detection areas at intervals, each detection area consists of five detection blocks 2 and six isolation blocks 3, the five isolation blocks 3 and the six detection blocks 2 are arranged at intervals,
the liquid absorbing zone 1 is made of a glass fiber film, the spacer 3 is made of a curable polymer,
the base plate 4 is arranged below the test paper, three through sample injection holes 6 are arranged in the center of the base plate 4, the three sample injection holes 6 respectively correspond to different liquid absorption areas 1,
baffle plates 5 are arranged upwards at the peripheral edges of the substrate 4, and the test paper is fixed on the substrate 4 by the baffle plates 5.
A preparation method of urine analysis test paper comprises the following steps:
(1) cutting a glass fiber film into 10mm multiplied by 7mm in size to serve as a test paper substrate, dripping a curable polymer onto the test paper substrate, and polymerizing and curing the curable polymer to form a separation block 3 serving as a hydrophobic barrier, and separating the test paper substrate into three rows of five detection blocks 2 each, wherein the periphery of each detection block 2 is surrounded by the hydrophobic barrier;
(2) respectively performing chemical reagent dripping and treatment on fifteen detection blocks 2, and respectively preparing the fifteen detection blocks 2 into project detection blocks for detecting different projects;
the project detection block is a glucose detection block, a PH value detection block, a protein detection block, a microalbumin detection block, a specific gravity detection block, a ketone detection block, a bilirubin detection block, a urobilinogen detection block, a nitrite detection block, a leucocyte detection block, an ascorbic acid detection block, a occult blood detection block, a uric acid detection block, a creatinine detection block and a calcium detection block, and the preparation method of each project detection block specifically comprises the following steps:
preparation of glucose test block
Weighing 0.19g of citric acid, adding into 10ml of ultrapure water, stirring thoroughly until the citric acid is completely dissolved, regulating the pH of the solution to 6.0 by using 0.1mol/L sodium hydroxide, adding 0.025g of tetramethylbenzidine, 920u of glucose oxidase and 19u of peroxidase into the buffer solution, and stirring until the citric acid is completely dissolved; spotting the prepared solution on a glucose detection area by using a pipette, wherein the sample size is 0.5 mu l, and drying at 25 ℃ after spotting is finished;
preparation of PH value detection block
1.2g of methyl red, 0.8g of bromothymol blue and 0.8ml of ethanol are weighed and added into 10ml of ultrapure water, and the mixture is fully stirred until the mixture is completely dissolved; spotting the prepared solution on the pH value detection area by using a pipette, wherein the sample size is 0.5 mu l, and drying at 50 ℃ after spotting is finished;
protein detection Block preparation
Weighing 0.095g of citric acid, adding into 10ml of ultrapure water, fully stirring until the citric acid is completely dissolved, regulating the pH of the solution to 2.0 by using 0.1mol/L of sodium hydroxide and 0.1mol/L of hydrochloric acid, adding 0.8ml of ethanol, 0.025g of polyvinyl alcohol and 0.033g of tetrabromophenol blue into the buffer solution, and stirring until the tetrabromophenol blue is completely dissolved; spotting the prepared solution on a protein detection area by using a pipette, wherein the sample size is 0.5 mu l, and drying at 50 ℃ after spotting is finished;
preparation of microalbumin detection block
1.9g of citric acid is weighed and added into 10ml of ultrapure water, the mixture is fully stirred until the mixture is completely dissolved, 1mol/L of sodium hydroxide and 1mol/L of hydrochloric acid are used for regulating the pH of the solution to 2.0, 0.01g of sulfophthalein dye is added into the buffer solution, and the mixture is stirred until the mixture is completely dissolved; spotting the prepared solution on a trace albumin detection area by using a pipette, wherein the sample size is 0.5 mu l, and drying at 50 ℃ after spotting is finished;
specific gravity detection block preparation
Weighing 0.012g of trimetaphosphate, 0.018g of bromothymol blue and 0.05g of tetramethylammonium hydroxide, adding into 10ml of ultrapure water, fully stirring until the solution is completely dissolved, using a pipette to sample a specific gravity detection area, sampling the solution with a sample size of 0.5 mu l, and drying at 50 ℃ after sample application;
preparation of ketone body detection block
Weighing 0.5g of sodium hydroxide, adding 0.125g of sodium nitrosoferricyanide into 10ml of ultrapure water, fully stirring until the sodium hydroxide is completely dissolved, using a pipettor to sample the ketone detection area with the prepared solution, and drying at 35 ℃ after sample application is finished, wherein the sample size is 0.5 μl;
bilirubin detection block preparation
Weighing 0.95g of citric acid, adding into 10ml of ultrapure water, fully stirring until the citric acid is completely dissolved, adjusting the pH of the solution to 5.0 by using 1mol/L sodium hydroxide, adding 0.35g of 2,6 dichloro diazonium salt fluoborate into the buffer solution, and stirring until the boric acid is completely dissolved; spotting the bilirubin detection area with a sample size of 0.5 μl by using a pipette, and drying at 50deg.C after spotting;
preparation of urine bilinogen detection block
Weighing 0.95g of citric acid, adding into 10ml of ultrapure water, fully stirring until the citric acid is completely dissolved, adjusting the pH of the solution to 5.0 by using 1mol/L sodium hydroxide, adding 0.35g of p-xylene diazonium boron tetrafluoride into the buffer solution, and stirring until the boron tetrafluoride is completely dissolved; sample application is carried out on the urine bilinogen detection area by using a liquid transfer device, the sample size is 0.5 mu l, and the sample is dried at 50 ℃ after the sample application is finished;
nitrite detection block preparation
0.08g of p-aminobenzene arsinic acid, 0.09g of N- (1-naphthylamine) -ethylenediamine and 0.01g of sodium periodate are weighed and added into 10ml of ultrapure water, and the mixture is fully stirred until the sodium periodate is completely dissolved; sample application is carried out on the nitrite detection area by using a pipette, the sample size is 0.5 mu l, and the sample is dried at 30 ℃ after the sample application is finished;
preparation of leucocyte detection blocks
Weighing 0.25g of disodium hydrogen phosphate, 0.03g of sodium dihydrogen phosphate and 0.09g of sodium chloride, adding into 10ml of ultrapure water, fully stirring until the disodium hydrogen phosphate, the sodium chloride and the sodium chloride are completely dissolved, adjusting the pH of a solution to 9.0 by using 0.1mol/L sodium hydroxide, adding 0.04g of pyrrole ester, 0.02g of diazonium salt and 0.075g of imidazole into the buffer solution, and stirring until the sodium hydrogen phosphate, the sodium dihydrogen phosphate and the sodium chloride are completely dissolved; spotting the prepared solution on a leukocyte detection area by using a pipette, wherein the sample size is 0.5 mu l, and drying at 50 ℃ after spotting is finished;
preparation of ascorbic acid detection block
Weighing 0.08g of 2, 6-dichlorophenol indophenol, adding into 10ml of ultrapure water, and fully stirring until the indophenol is completely dissolved; spotting the prepared solution on an ascorbic acid detection area by using a pipette, wherein the sample size is 0.5 mu l, and drying at 40 ℃ after spotting is finished;
preparation of occult blood detection block
Weighing 0.17g of disodium hydrogen phosphate, 0.03g of sodium dihydrogen phosphate and 0.09g of sodium chloride, adding into 10ml of ultrapure water, fully stirring until the disodium hydrogen phosphate, the sodium chloride and the sodium chloride are completely dissolved, adjusting the pH of a solution to 6.5 by using 0.1mol/L of sodium hydroxide and 0.1mol/L of hydrochloric acid, taking 0.012g of carbamide peroxide, 0.08g of o-tolidine, 0.05g of 2-aminothiazole acetate and 2.6g of potassium iodate, adding into the buffer solution, and stirring until the potassium hydrogen phosphate, the sodium hydrogen phosphate and the sodium chloride are completely dissolved; spotting the prepared solution on a occult blood detection area by using a pipette, wherein the sample size is 0.5 mu l, and drying at 30 ℃ after the spotting is finished;
uric acid detection block preparation
Weighing 0.20g of disodium hydrogen phosphate, 0.03g of sodium dihydrogen phosphate and 0.09g of sodium chloride, adding into 10ml of ultrapure water, fully stirring until the disodium hydrogen phosphate, the sodium chloride and the sodium chloride are completely dissolved, adjusting the pH of a solution to 7.2 by using 0.1mol/L of sodium hydroxide and 0.1mol/L of hydrochloric acid, adding 0.06g of 4-aminoantipyrine, 0.04g of 2-hydroxy-3, 5-dichlorobenzenesulfonic acid, 15u of uricase and 25u of peroxidase into the buffer solution, and stirring until the solution is completely dissolved; using a pipette or an automatic spotting instrument to spot the prepared solution on the uric acid detection area, wherein the sample size is 0.5 mu l, and drying at 25 ℃ after the spot is finished;
preparation of creatinine detection block
Weighing 0.12G boric acid, adding into 10ml ultrapure water, fully stirring until the boric acid is completely dissolved, regulating the pH of the solution to 12 by using 0.1mol/L sodium hydroxide and 0.1mol/L hydrochloric acid, adding 0.05G potassium chloride, 0.02G polyvinylpyrrolidone, 0.001G orange G and 0.005G 3.5-dinitrobenzoic acid into the buffer solution, and stirring until the boric acid is completely dissolved; spotting the prepared solution on a creatinine detection area by using a pipette, wherein the sample size is 0.5 mu l, and drying at 40 ℃ after spotting is finished;
preparation of calcium detection block
Weighing 0.19g of lemon yellow, adding into 10ml of ultrapure water, fully stirring until the lemon yellow is completely dissolved, regulating the pH of a solution to 9.5 by using 0.1mol/L sodium hydroxide and 0.1mol/L hydrochloric acid, taking 0.035g of o-cresolphthalein complexing ketone, adding 0.1ml of Triton x-100 and 0.035g of EGTA into the buffer solution, and stirring until the lemon yellow is completely dissolved; spotting the prepared solution on a creatinine detection area by using a pipette, wherein the sample size is 0.5 mu l, and drying at 60 ℃ after spotting is finished;
(3) fixing the prepared test paper in a substrate 4 with three sample injection holes 6 at the bottom;
example 3
As shown in fig. 1 and 2, the urine analysis test paper comprises a test paper and a substrate 4, wherein the test paper comprises three liquid absorbing areas 1 with rectangular strip shapes and four detection areas with rectangular strip shapes, the four liquid absorbing areas 1 are arranged with the three detection areas at intervals, each detection area consists of five detection blocks 2 and six isolation blocks 3, the five isolation blocks 3 and the six detection blocks 2 are arranged at intervals,
the liquid absorbing zone 1 is made of a glass fiber film, the spacer 3 is made of a curable polymer,
the base plate 4 is arranged below the test paper, three through sample injection holes 6 are arranged in the center of the base plate 4, the three sample injection holes 6 respectively correspond to different liquid absorption areas 1,
baffle plates 5 are arranged upwards at the peripheral edges of the substrate 4, and the test paper is fixed on the substrate 4 by the baffle plates 5.
A preparation method of urine analysis test paper comprises the following steps:
(1) cutting a water-absorbing material into 10mm multiplied by 7mm in size to serve as a test paper base material, dipping the test paper base material with a hydrophobic material drop, polymerizing and solidifying the test paper base material to form a separation block 3 serving as a hydrophobic barrier, and separating the test paper base material into three rows of four detection blocks 2 each, wherein the periphery of each detection block 2 is surrounded by the hydrophobic barrier;
(2) respectively performing chemical reagent dripping and treatment on fifteen detection blocks 2, and respectively preparing the fifteen detection blocks 2 into project detection blocks for detecting different projects;
the project detection block is a glucose detection block, a PH value detection block, a protein detection block, a microalbumin detection block, a specific gravity detection block, a ketone detection block, a bilirubin detection block, a urobilinogen detection block, a nitrite detection block, a leucocyte detection block, an ascorbic acid detection block, a occult blood detection block, a uric acid detection block, a creatinine detection block and a calcium detection block, and the preparation method of each project detection block specifically comprises the following steps:
preparation of glucose test block
Weighing 0.19g of citric acid, adding into 10ml of ultrapure water, stirring thoroughly until the citric acid is completely dissolved, regulating the pH of the solution to 6.0 by using 0.1mol/L sodium hydroxide, adding 0.025g of tetramethylbenzidine, 920u of glucose oxidase and 19u of peroxidase into the buffer solution, and stirring until the citric acid is completely dissolved; using an automatic sample application instrument to sample the prepared solution to a glucose detection area, wherein the sample size is 1 μl, and drying at 35 ℃ after sample application is completed;
preparation of PH value detection block
1.2g of methyl red, 0.8g of bromothymol blue and 0.8ml of ethanol are weighed and added into 10ml of ultrapure water, and the mixture is fully stirred until the mixture is completely dissolved; using an automatic sample application instrument to sample the prepared solution in the PH value detection area, wherein the sample size is 1 μl, and drying at 80 ℃ after sample application is completed;
protein detection Block preparation
Weighing 0.095g of citric acid, adding into 10ml of ultrapure water, fully stirring until the citric acid is completely dissolved, adjusting the pH of the solution to 3.0 by using 0.1mol/L of sodium hydroxide and 0.1mol/L of hydrochloric acid, adding 0.8ml of ethanol, 0.025g of polyvinyl alcohol and 0.033g of tetrabromophenol blue into the buffer solution, and stirring until the solution is completely dissolved; using an automatic sample application instrument to sample the prepared solution to a protein detection area, wherein the sample size is 1 μl, and drying at 80 ℃ after sample application is completed;
preparation of microalbumin detection block
1.9g of citric acid is weighed and added into 10ml of ultrapure water, the mixture is fully stirred until the mixture is completely dissolved, 1mol/L of sodium hydroxide and 1mol/L of hydrochloric acid are used for regulating the pH of the solution to 3.0, 0.01g of sulfophthalein dye is added into the buffer solution, and the mixture is stirred until the mixture is completely dissolved; using an automatic sample application instrument to sample the prepared solution to a trace albumin detection area, wherein the sample size is 1 μl, and drying at 70 ℃ after sample application is completed;
specific gravity detection block preparation
Weighing 0.012g of trimetaphosphate, 0.018g of bromothymol blue and 0.05g of tetramethylammonium hydroxide, adding into 10ml of ultrapure water, fully stirring until the solution is completely dissolved, spotting a specific gravity detection area by using an automatic spotting instrument, wherein the sample size is 1 μl, and drying at 80 ℃ after spotting;
preparation of ketone body detection block
Weighing 0.5g of sodium hydroxide, adding 0.125g of sodium nitrosoferricyanide into 10ml of ultrapure water, fully stirring until the sodium hydroxide is completely dissolved, spotting a ketone detection area by using an automatic spotting instrument by using the prepared solution, wherein the sample size is 1 μl, and drying at 50 ℃ after spotting is finished;
bilirubin detection block preparation
Weighing 0.95g of citric acid, adding into 10ml of ultrapure water, fully stirring until the citric acid is completely dissolved, adjusting the pH of the solution to 5.0 by using 1mol/L sodium hydroxide, adding 0.35g of 2, 6-dichloro diazonium salt fluoborate into the buffer solution, and stirring until the boric acid is completely dissolved; using an automatic sample application instrument to sample the bilirubin detection area with the prepared solution, wherein the sample size is 1 μl, and drying at 70 ℃ after sample application is completed;
preparation of urine bilinogen detection block
Weighing 0.95g of citric acid, adding into 10ml of ultrapure water, fully stirring until the citric acid is completely dissolved, adjusting the pH of the solution to 5.0 by using 1mol/L sodium hydroxide, adding 0.35g of p-xylene diazonium boron tetrafluoride into the buffer solution, and stirring until the boron tetrafluoride is completely dissolved; using an automatic sample application instrument to sample the prepared solution to the urine bilinogen detection area, wherein the sample size is 1 μl, and drying at 70 ℃ after sample application is completed;
nitrite detection block preparation
0.08g of p-aminobenzene arsinic acid, 0.09g of N- (1-naphthylamine) -ethylenediamine and 0.01g of sodium periodate are weighed and added into 10ml of ultrapure water, and the mixture is fully stirred until the sodium periodate is completely dissolved; using an automatic sample application instrument to sample the nitrite detection area with the prepared solution, wherein the sample size is 1 μl, and drying at 40 ℃ after sample application is completed;
preparation of leucocyte detection blocks
Weighing 0.25g of disodium hydrogen phosphate, 0.03g of sodium dihydrogen phosphate and 0.09g of sodium chloride, adding into 10ml of ultrapure water, fully stirring until the disodium hydrogen phosphate, the sodium chloride and the sodium chloride are completely dissolved, adjusting the pH of a solution to 9.0 by using 0.1mol/L sodium hydroxide, adding 0.04g of pyrrole ester, 0.02g of diazonium salt and 0.075g of imidazole into the buffer solution, and stirring until the sodium hydrogen phosphate, the sodium dihydrogen phosphate and the sodium chloride are completely dissolved; using an automatic sample application instrument to sample the prepared solution to the leukocyte detection area, wherein the sample size is 1 μl, and drying at 70deg.C after sample application;
preparation of ascorbic acid detection block
Weighing 0.08g of 2, 6-dichlorophenol indophenol, adding into 10ml of ultrapure water, and fully stirring until the indophenol is completely dissolved; using an automatic sample application instrument to sample the prepared solution to an ascorbic acid detection area, wherein the sample size is 1 μl, and drying at 60 ℃ after sample application is completed;
preparation of occult blood detection block
Weighing 0.17g of disodium hydrogen phosphate, 0.03g of sodium dihydrogen phosphate and 0.09g of sodium chloride, adding into 10ml of ultrapure water, fully stirring until the disodium hydrogen phosphate, the sodium chloride and the sodium chloride are completely dissolved, adjusting the pH of a solution to 6.5 by using 0.1mol/L of sodium hydroxide and 0.1mol/L of hydrochloric acid, taking 0.012g of carbamide peroxide, 0.08g of o-tolidine, 0.05g of 2-aminothiazole acetate and 2.6g of potassium iodate, adding into the buffer solution, and stirring until the potassium hydrogen phosphate, the sodium hydrogen phosphate and the sodium chloride are completely dissolved; using an automatic sample application instrument to sample the prepared solution on a occult blood detection area, wherein the sample size is 1 μl, and drying at 45 ℃ after sample application is completed;
uric acid detection block preparation
Weighing 0.20g of disodium hydrogen phosphate, 0.03g of sodium dihydrogen phosphate and 0.09g of sodium chloride, adding into 10ml of ultrapure water, fully stirring until the disodium hydrogen phosphate, the sodium chloride and the sodium chloride are completely dissolved, adjusting the pH of a solution to 7.2 by using 0.1mol/L of sodium hydroxide and 0.1mol/L of hydrochloric acid, adding 0.06g of 4-aminoantipyrine, 0.04g of 2-hydroxy-3, 5-dichlorobenzenesulfonic acid, 15u of uricase and 25u of peroxidase into the buffer solution, and stirring until the solution is completely dissolved; using an automatic sample application instrument to sample the prepared solution to the uric acid detection area, wherein the sample size is 1 μl, and drying at 35 ℃ after sample application is completed;
preparation of creatinine detection block
Weighing 0.12G boric acid, adding into 10ml ultrapure water, fully stirring until the boric acid is completely dissolved, regulating the pH of the solution to 12 by using 0.1mol/L sodium hydroxide and 0.1mol/L hydrochloric acid, adding 0.05G potassium chloride, 0.02G polyvinylpyrrolidone, 0.001G orange G and 0.005G 3.5-dinitrobenzoic acid into the buffer solution, and stirring until the boric acid is completely dissolved; using an automatic sample application instrument to sample the prepared solution to the creatinine detection region, wherein the sample size is 1 μl, and drying at 60deg.C after sample application is completed;
preparation of calcium detection block
Weighing 0.19g of lemon yellow, adding into 10ml of ultrapure water, fully stirring until the lemon yellow is completely dissolved, regulating the pH of a solution to 9.5 by using 0.1mol/L sodium hydroxide and 0.1mol/L hydrochloric acid, taking 0.035g of o-cresolphthalein complexing ketone, adding 0.1ml of Triton x-100 and 0.035g of EGTA into the buffer solution, and stirring until the lemon yellow is completely dissolved; using an automatic sample application instrument to sample the prepared solution to the creatinine detection region, wherein the sample size is 1 μl, and drying at 80deg.C after sample application is completed;
uric acid detection block preparation
Weighing 0.20g of disodium hydrogen phosphate, 0.03g of sodium dihydrogen phosphate and 0.09g of sodium chloride, adding into 10ml of ultrapure water, fully stirring until the disodium hydrogen phosphate, the sodium chloride and the sodium chloride are completely dissolved, adjusting the pH of a solution to 7.2 by using 0.1mol/L of sodium hydroxide and 0.1mol/L of hydrochloric acid, adding 0.06g of 4-aminoantipyrine, 0.04g of 2-hydroxy-3, 5-dichlorobenzenesulfonic acid, 15u of uricase and 25u of peroxidase into the buffer solution, and stirring until the solution is completely dissolved; using an automatic sample application instrument to sample the prepared solution to the uric acid detection area, wherein the sample size is 1 μl, and drying at 35 ℃ after sample application is completed;
preparation of creatinine detection block
Weighing 0.12G boric acid, adding into 10ml ultrapure water, fully stirring until the boric acid is completely dissolved, regulating the pH of the solution to 12 by using 0.1mol/L sodium hydroxide and 0.1mol/L hydrochloric acid, adding 0.05G potassium chloride, 0.02G polyvinylpyrrolidone, 0.001G orange G and 0.005G 3.5-dinitrobenzoic acid into the buffer solution, and stirring until the boric acid is completely dissolved; using an automatic sample application instrument to sample the prepared solution to the creatinine detection region, wherein the sample size is 1 μl, and drying at 60deg.C after sample application is completed;
preparation of calcium detection block
Weighing 0.19g of lemon yellow, adding into 10ml of ultrapure water, fully stirring until the lemon yellow is completely dissolved, regulating the pH of a solution to 9.5 by using 0.1mol/L sodium hydroxide and 0.1mol/L hydrochloric acid, taking 0.035g of o-cresolphthalein complexing ketone, adding 0.1ml of Triton x-100 and 0.035g of EGTA into the buffer solution, and stirring until the lemon yellow is completely dissolved; using an automatic sample application instrument to sample the prepared solution to the creatinine detection region, wherein the sample size is 1 μl, and drying at 80deg.C after sample application is completed;
(3) the prepared test paper is fixed in a base plate 4 with three sample injection holes 6 at the bottom.
When the invention is used, urine to be detected is added into the sample inlet 6 at the back of the substrate 4, a sample is distributed to the whole test paper through the capillary action of the liquid suction area 1, so that the liquid to be detected is immersed into each test block 2, and the surface of the test block 2 presents different patterns and colors through the reaction of the urine to be detected and the chemical reagent in the test block 2.
While the invention has been described above with reference to the accompanying drawings, it will be apparent that the invention is not limited to the above embodiments, but is intended to cover various modifications, either made by the method concepts and technical solutions of the invention, or applied directly to other applications without modification, within the scope of the invention.
Claims (1)
1. A preparation method of urine analysis test paper is characterized in that,
the urine analysis test paper comprises test paper and a substrate (4), wherein the test paper comprises a plurality of liquid absorbing areas (1) and detection areas which are in rectangular strip shapes, the liquid absorbing areas (1) and the detection areas are arranged in a spaced mode, each detection area consists of a plurality of detection blocks (2) and a plurality of isolation blocks (3), the isolation blocks (3) and the detection blocks (2) are arranged at intervals, the liquid absorbing areas (1) are made of water absorbing materials, the isolation blocks (3) are made of hydrophobic materials, the substrate (4) is padded below the test paper, the center of the substrate (4) is provided with transparent sample injection holes (6), three sample injection holes (6) are respectively corresponding to different liquid absorbing areas (1), the periphery of the substrate (4) is upwards provided with baffle plates (5), and the test paper is fixed on the substrate (4) by the baffle plates (5) to prepare a water absorbing material of the area (1) and a polyester fiber film; the hydrophobic material for preparing the isolation block (3) is one of a curable polymer and a photoresist;
the preparation method of the urine analysis test paper comprises the following specific steps:
(1) cutting a water-absorbing material into 10mm multiplied by 7mm serving as a test paper substrate, dipping the test paper substrate with a hydrophobic material drop, polymerizing and solidifying the test paper substrate to form a separation block (3) serving as a hydrophobic barrier, and separating the test paper substrate into a plurality of detection blocks (2), wherein the periphery of each detection block (2) is surrounded by the hydrophobic barrier;
(2) respectively performing chemical reagent dripping and treatment on the plurality of detection blocks (2), and respectively preparing the plurality of detection blocks (2) into project detection blocks for detecting different projects;
the project detection block is selected from a glucose detection block, a PH value detection block, a protein detection block, a microalbumin detection block, a specific gravity detection block, a ketone detection block, a bilirubin detection block, a urobilinogen detection block, a nitrite detection block, a leucocyte detection block, an ascorbic acid detection block, a occult blood detection block, a uric acid detection block, a creatinine detection block and a calcium detection block, and the preparation method of each project detection block specifically comprises the following steps:
preparation of glucose test block
Weighing 0.19g of citric acid, adding into 10ml of ultrapure water, stirring thoroughly until the citric acid is completely dissolved, regulating the pH of the solution to 6.0 by using 0.1mol/L sodium hydroxide, adding 0.025g of tetramethylbenzidine, 920u of glucose oxidase and 19u of peroxidase into the buffer solution, and stirring until the citric acid is completely dissolved; using a pipette or an automatic sample application instrument to sample the prepared solution to a glucose detection area, wherein the sample size is 0.5-1 μl, and drying at 25-35deg.C after sample application;
preparation of PH value detection block
1.2g of methyl red, 0.8g of bromothymol blue and 0.8ml of ethanol are weighed and added into 10ml of ultrapure water, and the mixture is fully stirred until the mixture is completely dissolved; using a pipettor or an automatic sample application instrument to sample the prepared solution in the pH value detection area, wherein the sample size is 0.5-1 μl, and drying at 50-80deg.C after sample application is completed;
protein detection Block preparation
Weighing 0.095g of citric acid, adding into 10ml of ultrapure water, fully stirring until the citric acid is completely dissolved, adjusting the pH of the solution to 2.0-3.0 by using 0.1mol/L of sodium hydroxide and 0.1mol/L of hydrochloric acid, adding 0.8ml of ethanol, 0.025g of polyvinyl alcohol and 0.033g of tetrabromophenol blue into the buffer solution, and stirring until the solution is completely dissolved; using a pipettor or an automatic sample application instrument to sample the prepared solution to a protein detection area, wherein the sample size is 0.5-1 μl, and drying at 50-80deg.C after sample application is completed;
preparation of microalbumin detection block
1.9g of citric acid is weighed and added into 10ml of ultrapure water, fully stirred until the citric acid is completely dissolved, the PH of the solution is regulated to 2.0 to 3.0 by using 1mol/L of sodium hydroxide and 1mol/L of hydrochloric acid, 0.01g of sulfophthalein dye is added into the buffer solution, and the solution is stirred until the solution is completely dissolved; using a pipette or an automatic sample application instrument to sample the prepared solution to a trace albumin detection area, wherein the sample size is 0.5-1 μl, and drying at 50-70deg.C after sample application;
specific gravity detection block preparation
Weighing 0.012g of trimetaphosphate, 0.018g of bromothymol blue and 0.05g of tetramethylammonium hydroxide, adding into 10ml of ultrapure water, fully stirring until the solution is completely dissolved, using a pipette or an automatic sample application instrument to sample the prepared solution in a specific gravity detection area, wherein the sample size is 0.5-1 mu l, and drying at 50-80 ℃ after the sample application is finished;
preparation of ketone body detection block
Weighing 0.5g of sodium hydroxide, adding 0.125g of sodium nitrosoferricyanide into 10ml of ultrapure water, fully stirring until the sodium hydroxide is completely dissolved, using a pipette or an automatic sample application instrument to sample the prepared solution in a ketone detection area, wherein the sample size is 0.5-1 μl, and drying at 35-50 ℃ after sample application is completed;
bilirubin detection block preparation
Weighing 0.95g of citric acid, adding into 10ml of ultrapure water, fully stirring until the citric acid is completely dissolved, adjusting the pH of the solution to 5.0 by using 1mol/L sodium hydroxide, adding 0.35g of 2,6 dichloro diazonium salt fluoborate into the buffer solution, and stirring until the boric acid is completely dissolved; using a pipettor or an automatic spotting instrument to sample the prepared solution in the bilirubin detection area, wherein the sample size is 0.5-1 μl, and drying at 50-70deg.C after the sample application is completed;
preparation of urine bilinogen detection block
Weighing 0.95g of citric acid, adding into 10ml of ultrapure water, fully stirring until the citric acid is completely dissolved, adjusting the pH of the solution to 5.0 by using 1mol/L sodium hydroxide, adding 0.35g of p-xylene diazonium boron tetrafluoride into the buffer solution, and stirring until the boron tetrafluoride is completely dissolved; using a pipettor or an automatic sample application instrument to sample the prepared solution to the urine bilinogen detection area, wherein the sample size is 0.5-1 μl, and drying at 50-70deg.C after sample application;
nitrite detection block preparation
0.08g of p-aminobenzene arsinic acid, 0.09g of N- (1-naphthylamine) -ethylenediamine and 0.01g of sodium periodate are weighed and added into 10ml of ultrapure water, and the mixture is fully stirred until the sodium periodate is completely dissolved; using a pipettor or an automatic sample application instrument to sample the prepared solution to a nitrite detection area, wherein the sample size is 0.5-1 μl, and drying at 30-40deg.C after sample application;
preparation of leucocyte detection blocks
Weighing 0.25g of disodium hydrogen phosphate, 0.03g of sodium dihydrogen phosphate and 0.09g of sodium chloride, adding into 10ml of ultrapure water, fully stirring until the disodium hydrogen phosphate, the sodium chloride and the sodium chloride are completely dissolved, adjusting the pH of a solution to 9.0 by using 0.1mol/L sodium hydroxide, adding 0.04g of pyrrole ester, 0.02g of diazonium salt and 0.075g of imidazole into the buffer solution, and stirring until the sodium hydrogen phosphate, the sodium dihydrogen phosphate and the sodium chloride are completely dissolved; using a pipette or an automatic spotting instrument to spot the prepared solution on a leukocyte detection area, wherein the sample size is 0.5-1 μl, and drying at 50-70deg.C after the spot is finished;
preparation of ascorbic acid detection block
Weighing 0.08g of 2, 6-dichlorophenol indophenol, adding into 10ml of ultrapure water, and fully stirring until the indophenol is completely dissolved; using a pipette or an automatic spotting instrument to spot the prepared solution on an ascorbic acid detection area, wherein the sample size is 0.5-1 ul, and drying at 40-60 ℃ after the spot is finished;
preparation of occult blood detection block
Weighing 0.17g of disodium hydrogen phosphate, 0.03g of sodium dihydrogen phosphate and 0.09g of sodium chloride, adding into 10ml of ultrapure water, fully stirring until the disodium hydrogen phosphate, the sodium chloride and the sodium chloride are completely dissolved, adjusting the pH of a solution to 6.5 by using 0.1mol/L of sodium hydroxide and 0.1mol/L of hydrochloric acid, taking 0.012g of carbamide peroxide, 0.08g of o-tolidine, 0.05g of 2-aminothiazole acetate and 2.6g of potassium iodate, adding into the buffer solution, and stirring until the potassium hydrogen phosphate, the sodium hydrogen phosphate and the sodium chloride are completely dissolved; using a pipettor or an automatic sample application instrument to sample the prepared solution to the occult blood detection area, wherein the sample size is 0.5-1 μl, and drying at 30-45deg.C after sample application;
uric acid detection block preparation
Weighing 0.20g of disodium hydrogen phosphate, 0.03g of sodium dihydrogen phosphate and 0.09g of sodium chloride, adding into 10ml of ultrapure water, fully stirring until the disodium hydrogen phosphate, the sodium chloride and the sodium chloride are completely dissolved, adjusting the pH of a solution to 7.2 by using 0.1mol/L of sodium hydroxide and 0.1mol/L of hydrochloric acid, adding 0.06g of 4-aminoantipyrine, 0.04g of 2-hydroxy-3, 5-dichlorobenzenesulfonic acid, 15u of uricase and 25u of peroxidase into the buffer solution, and stirring until the solution is completely dissolved; using a pipettor or an automatic sample application instrument to sample the prepared solution to the uric acid detection area, wherein the sample size is 0.5-1 μl, and drying at 25-35deg.C after sample application;
preparation of creatinine detection block
Weighing 0.12G boric acid, adding into 10ml ultrapure water, fully stirring until the boric acid is completely dissolved, regulating the pH of the solution to 12 by using 0.1mol/L sodium hydroxide and 0.1mol/L hydrochloric acid, adding 0.05G potassium chloride, 0.02G polyvinylpyrrolidone, 0.001G orange G and 0.005G 3.5-dinitrobenzoic acid into the buffer solution, and stirring until the boric acid is completely dissolved; using a pipettor or an automatic sample application instrument to sample the prepared solution in the creatinine detection area, wherein the sample size is 0.5-1 μl, and drying at 40-60deg.C after the sample application is completed;
preparation of calcium detection block
Weighing 0.19g of lemon yellow, adding into 10ml of ultrapure water, fully stirring until the lemon yellow is completely dissolved, regulating the pH of a solution to 9.5 by using 0.1mol/L sodium hydroxide and 0.1mol/L hydrochloric acid, taking 0.035g of o-cresolphthalein complexing ketone, adding 0.1ml of Triton x-100 and 0.035g of EGTA into the buffer solution, and stirring until the lemon yellow is completely dissolved; using a pipettor or an automatic sample application instrument to sample the prepared solution in the creatinine detection area, wherein the sample size is 0.5-1 μl, and drying at 60-80deg.C after sample application is completed;
(3) the prepared test paper is fixed in a base plate (4) with three holes at the bottom.
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| CN109254000A (en) * | 2018-10-25 | 2019-01-22 | 太原理工大学 | Array urine multiple determination apparatus and method based on smart machine colorimetric analysis |
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