CN113884686A - Siphon type total homocysteine test paper and manufacturing method thereof - Google Patents
Siphon type total homocysteine test paper and manufacturing method thereof Download PDFInfo
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Abstract
The invention discloses a siphon type total homocysteine test paper and a manufacturing method thereof, wherein the test paper structure comprises a hydrophilic cover plate, a PET double-sided adhesive blood guide groove, a polyester gauze, a blood sucking pad, a blood filtering film, a color developing film, a PET interlayer, a double-sided adhesive pad and a PET bottom plate from top to bottom in sequence; the invention can carry out the test by using the blood of the fingertip, the siphon type design of the siphon sample adding port can lead the test speed to be faster, the time to be saved, the whole design saves the space, the sampling is convenient and less, the patient does not suffer, the sampling of professionals is not depended, the test result covers all the homocysteine forms, the result sensitivity is higher and more accurate, the problem that the sum of all the homocysteine forms can not be detected when the homocysteine is detected at present is perfectly solved, the interference problem of the dissociating agent is solved, and the requirement of the primary hospital inspection place on the rapid and convenient detection is met and suitable.
Description
Technical Field
The invention relates to the technical field of test paper, in particular to siphon-type total homocysteine test paper and a manufacturing method thereof.
Background
Homocysteine (HCY) is a sulfur-containing amino acid, is an intermediate product of methionine metabolism, does not participate in protein synthesis per se, cannot synthesize homocysteine in vivo, can only be converted from methionine, cannot synthesize methionine in a human body, and must be supplied by food, the main factors influencing the homocysteine level are genetic and food nutrient deficiency, vitamin B6, vitamin B12 and folic acid are required in homocysteine metabolism, if the concentrations of vitamin B12, vitamin B6 and folic acid in blood are too low, the homocysteine concentration in blood is increased, which is commonly seen in atherosclerotic angiopathy, cerebral apoplexy, rheumatoid arthritis, various cancers and the like, the homocysteine level is increased, and superoxide and peroxide generated by the homocysteine can cause vascular endothelial cell injury and low-density lipoprotein oxidation to cause continuous contraction and hypoxia of vascular smooth muscle, thus accelerating the atherosclerotic process, it may also disrupt normal coagulation mechanisms, increasing the chance of thrombosis.
Aiming at the requirement of rapid and convenient detection of homocysteine, the existing test methods do not reflect the dissociation process of the conjugated HCY, so the detected HCY is only in a free state in blood, but the content of tHCY cannot be detected, but the free HCY only accounts for 1 percent of the tHCY, the amount of the free HCY is too small to feed back more useful information in terms of results, and the significance for clinical diagnosis is not great;
the patent (application No. CN202010147017.5) discloses a dry type homocysteine test card and application thereof, the method solves the technical problem that tHCY can not be measured by the prior detection method, in the scheme, a method of adding a dissociating agent to change all binding type HCY into free type is adopted, enzyme is added after the dissociation is finished, hydrogen peroxide is finally generated, under the catalysis of peroxidase, a Trinder's chromogenic substrate is oxidized by hydrogen peroxide to show color change, although the method enables the binding type homocysteine to be dissociated, the dissociating agent contains reducibility and can react with the hydrogen peroxide, so that the concentration of a final product is reduced, and the generation amount of the hydrogen peroxide is proportional to the homocysteine, so that the reduction of the hydrogen peroxide content can influence the detection result, the accuracy is reduced, and the operation is complicated and easy to make mistakes.
The conventional method for detecting the total homocysteine commonly uses a high performance liquid chromatography, an enzyme-linked immunosorbent assay, a chemiluminescence method, a biochemical method, a circulating enzyme method and the like, and currently, in the international market, two HCY reagents which can be used on a biochemical analyzer are circulating enzyme methods, one is a direct method (a hydrolase circulating enzyme method) and the other is an indirect method (a cystathionine circulating enzyme method), wherein the hydrolase circulating enzyme method has high specificity and strong interference resistance, can eliminate endogenous beta-cystathionine, is suitable for various biochemical instruments, products of the type are widely adopted by international companies, the cystathionine circulating enzyme method indirect method reagent is easily interfered by lipase and iron reagents, the indirect method has the advantages of low price, but also needs to be operated on large-scale equipment such as biochemical instruments, and the like, but the methods can not meet the requirements of primary hospital inspection sites on quick and convenient detection, therefore, the invention provides the siphon-type total homocysteine test paper and the manufacturing method thereof to solve the problems in the prior art.
Disclosure of Invention
Aiming at the problems, the invention aims to provide a siphon type total homocysteine test paper and a manufacturing method thereof, the test paper can be tested by using fingertip blood, the siphon type design of a siphon sample adding port can lead the test speed to be faster, the time to be saved, the whole design saves the space, the sampling is convenient and less, the patient is not painful, the sampling is not dependent on professionals, the test result covers all the total homocysteine forms, and the result sensitivity is higher and more accurate.
In order to achieve the purpose of the invention, the invention is realized by the following technical scheme: the utility model provides a total homocysteine test paper of hydrocone type, includes the PET bottom plate, PET bottom plate top is equipped with the color development membrane, color development membrane top is equipped with strains the blood membrane, it is equipped with the blood sucking pad to strain blood membrane top, one side that PET bottom plate top is close to straining the blood membrane is equipped with the double faced adhesive tape mat that distributes side by side with straining the blood membrane, double faced adhesive tape mat top is equipped with the PET interlayer that distributes side by side with the blood sucking pad, it is equipped with the polyester gauze to inhale blood pad top, polyester gauze top is equipped with the PET double faced adhesive tape that covers PET interlayer and polyester gauze and leads the blood groove, the PET is glued and is led the blood groove top and be covered with the hydrophilicity cover plate, the PET double faced adhesive tape is led the blood groove and is gone up and has all seted up the siphon sample application mouth on the hydrophilicity cover plate.
The further improvement lies in that: the structure of PET double faced adhesive tape blood conduction groove and hydrophilic cover plate is the same, the one side of keeping away from the PET interlayer on PET double faced adhesive tape blood conduction groove and the hydrophilic cover plate has all seted up the inspection hole.
The further improvement lies in that: the surface of the blood filtering membrane is subjected to sample application treatment through disulfide and phosphate, the blood sucking pad is subjected to dipping pretreatment through a disulfide dissociation agent, and the PET interlayer is subjected to hydrophilic treatment through hydrophilic treatment liquid.
The further improvement lies in that: the surface of the color developing membrane is subjected to sample application treatment by reaction liquid, and the reaction liquid is formed by mixing peroxidase, 4-aminoantipyrine, pyruvate oxidase, cystathionine-beta-lyase and cystathionine-beta-synthetase.
A manufacturing method of siphon type total homocysteine test paper comprises the following steps:
step one
Printing a PET bottom plate, a PET interlayer and a PET double-sided adhesive blood guide groove by using a PET mould according to the positioning and shading requirements;
step two
Preparing a hydrophilic treatment liquid, and performing hydrophilic treatment on the PET interlayer by using the hydrophilic treatment liquid;
step three
Firstly, cutting a color development film raw material into long strip-shaped color development films according to required sizes by using a cutting machine, carrying out sample application and drying treatment on the color development films by using reaction liquid after the cutting is finished, and then pasting the color development films on a printed PET bottom plate;
step four
Firstly, cutting a blood filtering membrane raw material into long-strip blood filtering membranes according to required sizes by using a cutting machine, then using a disulfide and a phosphate to perform sample application on the surfaces of the blood filtering membranes, drying after the sample application is finished, and then attaching the dried blood filtering membranes to a color developing membrane;
step five
Firstly, slitting a blood absorption pad raw material into strip-shaped blood absorption pads according to required sizes by using a slitting machine, then carrying out impregnation pretreatment on the blood absorption pads by using a disulfide dissociation agent, drying the soaked blood absorption pads, and then attaching the dried blood absorption pads to a blood filtering film;
step six
Firstly, cutting a raw material of a double-sided adhesive tape cushion into strip-shaped double-sided adhesive tape cushions according to required sizes by using a cutting machine, then adhering the double-sided adhesive tape cushions to a PET bottom plate side by side along with a blood filtering film, and then adhering a processed PET interlayer to the double-sided adhesive tape cushions;
step seven
Firstly, cutting a polyester gauze raw material into long-strip-shaped polyester gauze by using a splitting machine according to requirements, then pasting the polyester gauze on a blood sucking pad and a PET interlayer, then sequentially pasting a PET double-faced adhesive tape blood guide groove on the polyester gauze, and then pasting a hydrophilic cover plate on the PET double-faced adhesive tape blood guide groove to obtain a full-page test strip;
step eight
The whole test strip is cut into small strips by a long strip cutting machine and a small strip cutting machine, and the cut small strips are loaded into a reagent cylinder to finish the manufacture of the total homocysteine test paper.
The further improvement lies in that: in the third step, the reaction solution is prepared by mixing peroxidase, 4-aminoantipyrine, pyruvate oxidase, cystathionine-beta-lyase and cystathionine-beta-synthetase according to the weight ratio of 1:1:1:1: 1.
The further improvement lies in that: in the third step, the sample application and drying treatment comprises the following specific steps: the prepared reaction solution is uniformly dripped on a color developing film, and then the color developing film is placed in a low-temperature tunnel furnace to be dried, the drying temperature is set to be 37 ℃, and the drying time is set to be 25 minutes.
The invention has the beneficial effects that: the invention can carry out the test by using the blood of the fingertip, the siphon type design of the siphon sample adding port can lead the test speed to be faster, the time to be saved, the whole design saves the space, the sampling is convenient and less, the patient does not suffer, the sampling of professionals is not depended, the test result covers all the homocysteine forms, the result sensitivity is higher and more accurate, the problem that the sum of all the homocysteine forms can not be detected when the homocysteine is detected at present is perfectly solved, the interference problem of the dissociating agent is solved, and the requirement of the primary hospital inspection place on the rapid and convenient detection is met and suitable.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to these drawings without creative efforts.
FIG. 1 is a schematic diagram of an overall structure of a test strip according to a first embodiment of the present invention;
FIG. 2 is a schematic diagram of a layered structure of the test paper according to the first embodiment of the present invention;
FIG. 3 is a schematic diagram of a side view of the test strip of the first embodiment of the present invention;
FIG. 4 is a standard curve of the total homocysteine biochemical concentration and signal value in example two of the present invention.
Wherein: 1. a PET bottom plate; 2. a color developing film; 3. a blood filtration membrane; 4. a blood suction pad; 5. a double-sided adhesive tape cushion; 6. a PET barrier layer; 7. polyester gauze; 8. a PET double-sided adhesive blood guide groove; 9. a hydrophilic cover sheet; 10. a siphon sample adding port; 11. and (6) detecting holes.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the description of the present invention, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc., indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, and are only for convenience of description and simplicity of description, but do not indicate or imply that the device or element being referred to must have a particular orientation, be constructed and operated in a particular orientation, and thus, should not be construed as limiting the present invention. Furthermore, the terms "first," "second," "third," "fourth," and the like are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.
In the description of the present invention, it should be noted that, unless otherwise explicitly specified or limited, the terms "mounted," "connected," and "connected" are to be construed broadly, e.g., as meaning either a fixed connection, a removable connection, or an integral connection; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meanings of the above terms in the present invention can be understood in specific cases to those skilled in the art.
Example one
Refer to fig. 1, 2, 3, this embodiment provides a total homocysteine test paper of hydrocone type, including PET bottom plate 1, 1 top of PET bottom plate is equipped with color development membrane 2, 2 tops of color development membrane are equipped with and strain blood membrane 3, it is equipped with blood sucking pad 4 to strain 3 tops of blood membrane, one side that 1 top of PET bottom plate is close to and strains blood membrane 3 is equipped with the double faced adhesive tape mat 5 that distributes side by side with straining blood membrane 3, 5 tops of double faced adhesive tape mat are equipped with the PET interlayer 6 that distributes side by side with blood sucking pad 4, 4 tops of blood sucking pad are equipped with polyester gauze 7, 7 tops of polyester gauze are equipped with the PET double faced adhesive tape that covers PET interlayer 6 and polyester gauze 7 and lead blood groove 8, 8 tops of PET double faced adhesive tape lead blood groove are covered with hydrophilicity cover plate 9, siphon sample application port 10 has all been seted up on PET double faced adhesive tape lead blood groove 8 and the hydrophilicity cover plate 9.
The structure that PET double faced adhesive tape led blood groove 8 and hydrophilic cover plate 9 is the same, and detection hole 11 has all been seted up to one side of keeping away from PET interlayer 6 on PET double faced adhesive tape led blood groove 8 and the hydrophilic cover plate 9.
The surface of the blood filtering membrane 3 is subjected to sample application treatment by disulfide and phosphate, the blood sucking pad 4 is subjected to impregnation pretreatment by a disulfide dissociation agent, and the PET interlayer 6 is subjected to hydrophilic treatment by hydrophilic treatment liquid.
The surface of the color developing film 2 is subjected to sample application treatment by reaction liquid, and the reaction liquid is formed by mixing peroxidase, 4-aminoantipyrine, pyruvate oxidase, cystathionine-beta-lyase and cystathionine-beta-synthetase.
The embodiment also provides a manufacturing method of the siphon-type total homocysteine test paper, which comprises the following steps:
step one
Printing a PET bottom plate 1, a PET interlayer 6 and a PET double-sided adhesive blood guide groove 8 by using a PET mould according to the positioning and shading requirements;
step two
Preparing a hydrophilic treatment liquid, and carrying out hydrophilic treatment on the PET interlayer 6 by using the hydrophilic treatment liquid;
step three
Firstly, cutting a color developing film raw material into long-strip color developing films 2 according to required sizes by using a splitting machine, carrying out sample application and drying treatment on the color developing films 2 by using reaction liquid after the cutting is finished, then pasting the color developing films 2 on a printed PET bottom plate 1, wherein the reaction liquid is prepared by mixing peroxidase, 4-aminoantipyrine, pyruvate oxidase, cystathionine-beta-lyase and cystathionine-beta-synthetase in a weight ratio of 1:1:1:1:1, and the specific steps of the sample application and drying treatment are as follows: firstly, uniformly dripping prepared reaction liquid on a color development membrane 2, then putting the color development membrane 2 into a low-temperature tunnel furnace for drying, setting the drying temperature at 37 ℃ and the drying time at 25 minutes;
step four
Firstly, cutting a blood filtering membrane raw material into long strips of blood filtering membranes 3 according to required sizes by using a cutting machine, then using a disulfide and a phosphate to perform sample application on the surfaces of the blood filtering membranes 3, drying after the sample application is finished, and then attaching the dried blood filtering membranes 3 to a color developing membrane 2;
step five
Firstly, slitting a blood absorption pad raw material into strip-shaped blood absorption pads 4 according to required sizes by using a slitting machine, then carrying out impregnation pretreatment on the blood absorption pads 4 by using a disulfide dissociation agent, drying after the impregnation is finished, and then attaching the dried blood absorption pads 4 to a blood filtering membrane 3;
step six
Firstly, cutting a raw material of the double-sided adhesive tape cushion into strip-shaped double-sided adhesive tape cushions 5 according to required sizes by using a cutting machine, then adhering the double-sided adhesive tape cushions 5 to the PET bottom plate 1 side by side next to the blood filtering film 3, and then adhering the processed PET interlayer 6 to the double-sided adhesive tape cushions 5;
step seven
Firstly, cutting a polyester gauze raw material into long-strip-shaped polyester gauze 7 according to requirements by using a splitting machine, then pasting the polyester gauze 7 on a blood sucking pad 4 and a PET interlayer 6, then sequentially pasting a PET double-faced adhesive tape blood guide groove 8 on the polyester gauze 7, and then pasting a hydrophilic cover plate 9 on the PET double-faced adhesive tape blood guide groove 8 to obtain a whole-plate test strip;
step eight
The whole test strip is cut into small strips by a long strip cutting machine and a small strip cutting machine, and the cut small strips are loaded into a reagent cylinder to finish the manufacture of the total homocysteine test paper.
The reaction mechanism is as follows: blood is sucked from a siphon port sample adding port 10, and is uniformly diffused to a blood sucking pad 4 containing a disulfide dissociation agent through a polyester gauze 7, wherein a combined HCY can be dissociated, then the combined HCY permeates downwards to a blood filtering membrane 3, the blood filtering membrane 3 treated by disulfide and phosphate eliminates an excessive dissociation agent, red blood cells are filtered, finally, the free total homocysteine (tHCY) generates a color development reaction on a color development membrane 2, the absorbance of a color development area is detected through a detection hole 11 to quantitatively detect the total homocysteine (tHCY), the instrument detects the color development concentration, and the total homocysteine concentration is inversely proportional to the color development concentration.
Example two
Taking 5 parts of fresh sample whole blood with gradient total homocysteine concentration, wherein each concentration is divided into 3 parts, the first part is subjected to total homocysteine test by biochemical analysis, the second part is subjected to total homocysteine standard curve determination, the third part is subjected to total homocysteine test paper test, the obtained result is compared with the biochemical analysis, and the obtained test data and the standard curve are shown in the following table 1:
TABLE 1
The experimental data is standard curve confirmation, and the experimental data shows that the tested signal value has good repeatability and the detected signal is a reflectance signal, so that the higher the concentration of the total homocysteine is, the more hydrogen peroxide is generated by reaction, the deeper the concentration of the chromogenic reaction is, the less the reflectance is, and the smaller the signal value is; according to the experimental data, a standard curve of the total homocysteine biochemical concentration and the signal value is linearly fitted, as shown in fig. 4 (the standard curve represents the linear relationship between the reaction signal and the concentration of a batch of test strips, also called CODE, such as the formula y ═ kx + b), in the graph, the vertical axis of the curve represents the total homocysteine concentration, and the horizontal axis represents the signal value, and as can be seen from the standard curve, the linear correlation is good and can reach 0.9915.
The third sample was tested using the total homocysteine test paper, and the test results are shown in table 2 below:
TABLE 2
The test result shows that the total homocysteine test paper has good repeatability, the CV is within 5 percent, and the deviation from the biochemical value tested by a large-scale biochemical instrument is small, which indicates that the total homocysteine test paper has high accuracy, thus completely solving the problems of low sensitivity and insufficient accuracy of the current total homocysteine test paper on a test sample and greatly reducing the complexity and the cost of the test.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (7)
1. The utility model provides a total homocysteine test paper of hydrocone type, includes PET bottom plate (1), its characterized in that: the top end of the PET bottom plate (1) is provided with a color development film (2), the top end of the color development film (2) is provided with a blood filtration film (3), a blood sucking pad (4) is arranged at the top end of the blood filtering membrane (3), a double-faced adhesive tape pad (5) which is distributed with the blood filtering membrane (3) side by side is arranged at one side of the top end of the PET bottom plate (1) close to the blood filtering membrane (3), the top end of the double-sided adhesive tape cushion (5) is provided with a PET interlayer (6) which is distributed with the blood absorption cushion (4) side by side, the top end of the blood sucking pad (4) is provided with a polyester gauze (7), the top end of the polyester gauze (7) is provided with a PET double-sided adhesive tape blood guide groove (8) which covers the PET interlayer (6) and the polyester gauze (7), the top end of the PET double-sided adhesive blood guide groove (8) is covered with a hydrophilic cover sheet (9), siphon sample adding openings (10) are formed in the PET double-sided adhesive blood guide groove (8) and the hydrophilic cover plate (9).
2. The siphonic total homocysteine test strip according to claim 1 wherein: the PET double faced adhesive tape blood guide groove (8) and the hydrophilic cover plate (9) are identical in structure, and the detection holes (11) are formed in one sides of the PET double faced adhesive tape blood guide groove (8) and the hydrophilic cover plate (9) far away from the PET interlayer (6).
3. The siphonic total homocysteine test strip according to claim 1 wherein: the surface of the blood filtering membrane (3) is subjected to sample application treatment through disulfide and phosphate, the blood sucking pad (4) is subjected to impregnation pretreatment through a disulfide dissociator, and the PET interlayer (6) is subjected to hydrophilic treatment through hydrophilic treatment liquid.
4. The siphonic total homocysteine test strip according to claim 1 wherein: the surface of the color developing membrane (2) is subjected to sample application treatment by reaction liquid, and the reaction liquid is formed by mixing peroxidase, 4-aminoantipyrine, pyruvate oxidase, cystathionine-beta-lyase and cystathionine-beta-synthetase.
5. A manufacturing method of siphon type total homocysteine test paper is characterized by comprising the following steps:
step one
Printing a PET bottom plate (1), a PET interlayer (6) and a PET double-sided adhesive blood guide groove (8) by using a PET mould according to the positioning and shading requirements;
step two
Preparing hydrophilic treatment liquid, and then carrying out hydrophilic treatment on the PET interlayer (6) by using the hydrophilic treatment liquid;
step three
Firstly, cutting a developing film raw material into long-strip developing films (2) according to required sizes by using a cutting machine, carrying out sample application and drying treatment on the developing films (2) by using reaction liquid after the cutting is finished, and then attaching the developing films (2) to a printed PET bottom plate (1);
step four
Firstly, cutting a blood filtering membrane raw material into long-strip-shaped blood filtering membranes (3) according to required sizes by using a splitting machine, then using disulfide and phosphate to perform sample application on the surfaces of the blood filtering membranes (3), drying after the sample application is finished, and then attaching the dried blood filtering membranes (3) to a color developing membrane (2);
step five
Firstly, slitting a blood absorption pad raw material into strip-shaped blood absorption pads (4) according to required sizes by using a slitting machine, then carrying out impregnation pretreatment on the blood absorption pads (4) by using a disulfide dissociation agent, drying after impregnation is finished, and then attaching the dried blood absorption pads (4) to a blood filtering membrane (3);
step six
Firstly, cutting a raw material of the double-sided adhesive tape cushion into strip-shaped double-sided adhesive tape cushions (5) according to required sizes by using a cutting machine, then adhering the double-sided adhesive tape cushions (5) to a PET bottom plate (1) side by side along with a blood filtering film (3), and then adhering a processed PET interlayer (6) to the double-sided adhesive tape cushions (5);
step seven
Firstly, cutting a polyester gauze raw material into long-strip-shaped polyester gauze (7) by using a splitting machine according to requirements, then pasting the polyester gauze (7) on a blood sucking pad (4) and a PET interlayer (6), then sequentially pasting a PET double-sided adhesive blood guide groove (8) on the polyester gauze (7), and then pasting a hydrophilic cover plate (9) on the PET double-sided adhesive blood guide groove (8) to obtain a full-page test strip;
step eight
The whole test strip is cut into small strips by a long strip cutting machine and a small strip cutting machine, and the cut small strips are loaded into a reagent cylinder to finish the manufacture of the total homocysteine test paper.
6. The method of claim 5, wherein the test strip is prepared by a method comprising the steps of: in the third step, the reaction solution is prepared by mixing peroxidase, 4-aminoantipyrine, pyruvate oxidase, cystathionine-beta-lyase and cystathionine-beta-synthetase according to the weight ratio of 1:1:1:1: 1.
7. The method of claim 5, wherein the test strip is prepared by a method comprising the steps of: in the third step, the sample application and drying treatment comprises the following specific steps: dripping the prepared reaction liquid on the color developing film (2) uniformly, putting the color developing film (2) into a low-temperature tunnel furnace for drying, setting the drying temperature at 37 ℃ and the drying time at 25 minutes.
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