CN113267628A - Sperm SP10 protein detection test strip and quantitative detection method - Google Patents
Sperm SP10 protein detection test strip and quantitative detection method Download PDFInfo
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- CN113267628A CN113267628A CN202110448949.8A CN202110448949A CN113267628A CN 113267628 A CN113267628 A CN 113267628A CN 202110448949 A CN202110448949 A CN 202110448949A CN 113267628 A CN113267628 A CN 113267628A
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Classifications
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- G—PHYSICS
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- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
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- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
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- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
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- G—PHYSICS
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Abstract
The invention provides a sperm SP10 protein detection test strip and a quantitative detection method, belonging to the technical field of instant detection. The technical scheme is as follows: the detection test strip is arranged in a lateral flow mode, a substrate is used as a substrate, and the structure on the substrate sequentially comprises a sample pad, a marking pad, a chromatographic membrane and a water absorption pad; the chromatographic membrane is provided with a detection area and a quality control area, and the labeling pad is attached with a functional nano material which can be combined with SP10 protein; two adjacent structures are partially overlapped. The invention has the beneficial effects that: the test strip is used for quantitatively detecting the concentration of male sperms, the sample solution containing SP10 protein is directly dripped onto the sample pad, and the quantitative detection of the concentration of SP10 protein is realized through the signal acquisition and analysis of the detection area.
Description
Technical Field
The invention relates to the technical field of instant detection, in particular to a sperm SP10 protein detection test strip and a quantitative detection method.
Background
According to the human semen examination and treatment laboratory manual, the main factors causing male infertility include semen amount, total number of sperms, semen concentration, semen normal form rate, semen pH value and liquefaction time, wherein the semen concentration is an important index for measuring male infertility, and when the semen concentration is less than 15 × 106When the number is one/mL, the growth is considered to be difficult. Based on anatomical, genetic and biochemical features, the SP10 protein has been shown to be useful as a biomarker indicative of male sperm concentration, which is expressed only in the testis, a marker differentiated in the last step of the spermatogenic lineage. According to the research, the concentration of the SP10 protein and the concentration of the sperm present a certain linear relationship, namely the concentration of the sperm can be quantitatively analyzed by detecting the concentration of the SP10 protein.
The lateral flow immunochromatography detection technology is a commercial technology developed in the 70 th generation of the 20 th century, and is widely applied to the field of instant detection until now, and particularly has wide application prospects in places with scarce resources. The existing SP10 protein detection Test paper strip (David Sperm SP10 protein detection Test paper, Jinxiu Sperm SP10 protein detection reagent, Fertili MARQ Fertility Sperm Test Kit, Spermcheck Fertility Home Test Kit and baby Fertility count Male Fertility Test) is identified based on a colorimetric method, only can identify SP10 protein, cannot accurately quantify the concentration of SP10 protein qualitatively, and further influences the accurate detection of Sperm concentration; in addition, the single sale price of the existing foreign commercialized test paper strip is also about seventy yuan, and the single sale price of the domestic product is about forty yuan, so that the existing commercialized test paper strip is generally higher in sale price and is not beneficial to popularization and use in poor places.
SERS has been widely used in the fields of environmental science, material analysis, food safety, and other analyses since 1974, and is a phenomenon in which the raman signal intensity of a raman dye molecule adsorbed on the surface of a metal nanomaterial is greatly enhanced. SERS has a single-molecule detection level and is a very sensitive detection method. Fluorescence detection refers to that after being irradiated by light, fluorescent molecules absorb and store light energy and enter an excited state, but the excited state is unstable, when the energy level of the fluorescent molecules returns to a ground state from the excited state, the excess energy is released in the form of electromagnetic radiation, and the released energy is fluorescence. Magnetic signal detection is a new detection method appearing in recent years, and the magnetic signal detection instrument is used for collecting magnetic signals of magnetic nanoparticles in a detection area so as to achieve the purpose of analyzing the target concentration in the detection area, and is applied to the field of molecular detection.
How to solve the above technical problems is the subject of the present invention.
Disclosure of Invention
The invention aims to provide a sperm SP10 protein detection test strip and a quantitative detection method, and discloses a sperm SP10 protein detection test strip which can simultaneously meet the requirements of low cost, simple and convenient operation, high sensitivity and the like, wherein the liquid flow mode of the test strip is lateral flow, the quantitative detection of SP10 protein is realized by using a nano material coded by a signal label, and the test strip is suitable for instant detection and has the advantages of simple and convenient operation, rapid detection, low cost and the like.
In order to achieve the purpose, the invention adopts the following technical scheme: a sperm SP10 protein detection test strip is designed in a lateral flow mode and comprises a substrate, a sample pad, a marking pad, a chromatographic membrane and a water absorption pad, wherein the substrate is used as a substrate and is positioned above the substrate, the sample pad, the marking pad, the chromatographic membrane and the water absorption pad are sequentially arranged from left to right, one end of the chromatographic membrane is arranged on the lower surface of the marking pad, and the upper surface of the other end of the chromatographic membrane is attached to the lower surface of the water absorption pad; the chromatographic membrane is also provided with a detection area and a quality control area; the labeling pad is attached with a functionalized nano material capable of being combined with SP10 protein.
The substrate, the sample pad, the marking pad, the chromatographic membrane and the water absorption pad are overlapped in sequence in a staggered manner, and the overlapping length is 2-4mm, so that the flowing continuity of sample liquid flow is ensured. In the specific practical application process of the test strip, a sample solution containing SP10 protein is directly dripped onto a sample pad, and the SP10 protein concentration is quantitatively detected through signal acquisition and analysis of a detection area.
As a further optimization scheme of the sperm SP10 protein detection test strip provided by the invention, the substrate is an inert hard material for preventing liquid from permeating, such as a PVC plate, a PDMS plate or a PS plate, the length of the substrate is 40-198mm, and the width of the substrate is 2-6 mm.
The sample pad is a glass fiber membrane, a nylon membrane, a non-woven membrane or a cellulose membrane, the length of the sample pad is 10-40mm, and the width of the sample pad is 2-6 mm.
The marking pad is a glass fiber film, a non-woven fabric, a cellulose film or a polyvinyl acetate film, the length of the marking pad is 6-10mm, and the width of the marking pad is 2-6 mm.
The chromatographic membrane is a nitrocellulose membrane, cellulose acetate, polyvinylidene fluoride or nylon membrane, and has a length of 20-80mm and a width of 2-6 mm.
The absorbent pad is cellulose membrane, absorbent paper, cellulose-glass fiber composite, filter paper, sponge or non-woven fabric, and has a length of 10-80mm and a width of 2-6 mm.
As a further optimized scheme of the sperm SP10 protein detection test strip provided by the invention, the detection area is an array consisting of a straight line, a single point or a plurality of points drawn on a chromatographic membrane by a reagent combined with SP10 protein, and the reagent combined with SP10 protein is a capture antibody, an aptamer or a ligand of SP 10.
As a further optimization scheme of the sperm SP10 protein detection test strip provided by the invention, the quality control area is an array consisting of a straight line, a single point or a plurality of points drawn on a chromatographic membrane by a reagent combined with a functionalized nano material.
The reagent combined with the functionalized nanometer material is a capture antibody, an aptamer or a ligand corresponding to the SP10 protein detection substance, and the capture antibody, the aptamer or the ligand corresponding to the SP10 protein detection substance is specifically combined with the SP10 protein detection substance and does not have cross reaction with the SP10 protein; the quality control area is positioned at the right side of the detection area.
As a further optimization scheme of the sperm SP10 protein detection test strip provided by the invention, a signal label is modified on a functionalized nano material combined with SP10 protein, and SP10 protein detection substances are fixed on the surface of the nano material through electrostatic adsorption or covalent connection, and the scheme specifically comprises the following steps: antibody, aptamer or ligand for quantitative detection of SP10 protein.
Or the nano material is not modified with any signal label, and a detection substance corresponding to the SP10 protein is fixed on the nano material, and specifically comprises the following components: an antibody, aptamer or ligand.
As a further optimized scheme of the sperm SP10 protein detection test strip provided by the invention, the nano-materials are as follows:
metal nanoparticles, including two types: the first type is a single-structure nanoparticle, specifically: gold, silver or copper nanoparticles having a particle size of 5-100 nm;
the second type is core-shell structured nanoparticles, which specifically comprise: silver core gold shell, gold core silver shell, gold core gold shell, silver core silver shell, silica core gold shell, silica core silver shell, gold core silica shell or silver core silica shell nanoparticles, wherein the size of the core is 2-80nm, and the thickness of the shell is 1.5-10 nm;
the magnetic nano material comprises the following specific components: ferroferric oxide, ferric oxide or manganese zinc ferrite nano-particles with the particle size of 5-100 nm;
③ the quantum dots specifically comprise: lead sulfide, indium phosphide/zinc sulfide or cadmium selenide/zinc sulfide quantum dots with the particle size of 5-100 nm;
the carbon nano material specifically comprises the following components: carbon nano-tubes or carbon nanospheres with a particle size of 5-100 nm;
as a further optimization scheme of the sperm SP10 protein detection test strip provided by the invention, the signal label is as follows:
the Raman dye molecule specifically comprises: p-aminophenol, p-mercaptobenzoic acid, methylene blue, rhodamine 6G, Nailan A, crystal violet, malachite green isothiocyanate or p-mercaptothiophenol;
② fluorescent dye molecules, specifically: cy3, Cy5, fluorescein isothiocyanate or phycoerythrin chlorophyll protein.
The signal label and the nano material are modified by electrostatic adsorption or covalent connection of the signal label to the surface of the nano material or the nuclear shell gap of the nuclear shell structure nano particle.
In order to better achieve the above object, the present invention further provides a quantitative detection method of the sperm SP10 protein detection test strip, comprising the following steps:
s1, directly dripping the sample solution containing the SP10 protein on the sample pad for 5-20 min;
and S2, carrying out signal acquisition analysis on the detection area and the quality control area, and carrying out quantitative detection on the SP10 protein.
As a further optimized scheme of the quantitative detection method of the sperm SP10 protein detection test strip provided by the invention, the signal is a fluorescence, Surface Enhanced Raman Scattering (SERS) or magnetic signal.
Compared with the prior art, the invention has the beneficial effects that:
1. the invention is used for quantitatively detecting the concentration of male sperms, the concentration of the sperms is used as an index for auxiliary diagnosis and curative effect observation of male infertility, and the test strip has an important function in male reproduction, the test strip is designed in a lateral flow mode, a substrate is used as a substrate, and the structures on the substrate sequentially comprise a sample pad, a marking pad, a chromatographic membrane and a water absorption pad, wherein the chromatographic membrane is provided with a detection area and a quality control area, the marking pad is attached with a functional nano material capable of being combined with SP10 protein, two adjacent structures are partially overlapped to ensure the continuity of the flow of a sample liquid.
2. The sperm SP10 protein detection test strip provided by the invention can realize quantitative detection of SP10 protein, so that the concentration of sperm can be more accurately judged.
3. The sperm SP10 protein detection test strip provided by the invention has strong specificity, and after the lateral flow of liquid flow, immunoreaction occurs in a detection area and a quality control area, the result is accurate, and the probability of false positive and false negative is lower.
4. The sperm SP10 protein detection test strip provided by the invention is user-friendly, and signal collection and analysis can be carried out on the detection area and the quality control area after 5-20min by only dripping the sample solution containing SP10 protein on the sample pad.
Drawings
FIG. 1 is a schematic composition diagram of a sperm SP10 protein test strip provided by the present invention.
Wherein the reference numerals are: 1. a substrate; 2. a sample pad; 3. a marking pad; 4. a chromatographic membrane; 5. a water absorbent pad; 6. a detection zone; 7. a quality control region; 8. a functionalized nanoparticle; 9. a sample containing SP10 protein; 10. forming a complex by the functionalized nanometer material, the SP10 protein and the capture substance in the detection area; 11. and a complex formed by the functionalized nanometer material and the capture substance in the quality control area.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. Of course, the specific embodiments described herein are merely illustrative of the invention and are not intended to be limiting.
Example 1
Referring to fig. 1, the invention provides a technical scheme that a sperm SP10 protein quantitative detection test strip based on SERS comprises the following steps:
A) construction of lateral flow test strips: the substrate 1 is made of a PVC plate, the length of the PVC plate is 40mm, and the width of the PVC plate is 2 mm; a sample pad 2, a marking pad 3, a chromatographic membrane 4 and a water absorption pad 5 are sequentially arranged on a substrate, wherein the sample pad is made of a glass fiber membrane, the length of the sample pad is 10mm, and the width of the sample pad is 2 mm; the marking pad is made of a glass fiber film, the length of the glass fiber film is 6mm, and the width of the glass fiber film is 2 mm; the chromatography membrane is made of a nitrocellulose membrane, and has a length of 20mm and a width of 2 mm; the water absorption pad is made of water absorption paper, the length of the water absorption pad is 10mm, and the width of the water absorption pad is 2 mm; the detection area 6 is a detection line formed by using SP10 capture antibody; the quality control region 7 is a quality control line drawn by using a capture antibody bound to the SP10 detection antibody. The quality control line is arranged at the right side of the detection line, the functionalized nano material 8 is fixed on the marking pad, the sample pad and the marking pad, the marking pad and the chromatographic membrane, and the chromatographic membrane and the water absorption pad are partially overlapped, and the overlapping length is 2 mm.
B) Preparing an SERS nano label: selecting 5nm gold nanoparticles as a SERS enhanced substrate, modifying a Delrin A Raman dye molecule to the surface of the gold nanoparticles in an electrostatic adsorption mode, and then fixing a detection antibody of SP10 protein to the surface of the gold nanoparticles in a covalent connection mode to finally form a functionalized nano material, namely an SERS nano label.
C) The use flow of the test strip for detecting the SP10 protein comprises the following steps:
firstly, a sample solution 9 containing SP10 protein is dropped on a sample pad, and the sample solution flows laterally due to capillary force. When the target SP10 protein reaches the marking pad, the target SP10 protein reacts with the SERS nano-label to form an SP10 protein-nano particle complex; with the further flow of the liquid flow, the SP10 protein-nanoparticle complex will be combined with the capture antibody on the detection line to form a sandwich structure 10; when the liquid flow reaches the control line, the functionalized nanoparticles will bind to the capture antibody on the control line to form a complex 11.
And secondly, after 5min, using a handheld Raman spectrometer to collect signals of the detection line and the quality control line on the test strip.
D) And (4) analyzing results:
quality control line detection: if a characteristic peak of the Narlan A appears in the Raman spectrum collected from the quality control line, the test strip is effective; otherwise, the test strip is invalid.
Detection line detection: if a characteristic peak of the Narland A appears in a Raman spectrum collected from the detection line, the existence of SP10 protein in the detection sample is indicated, and at the moment, the amount of the target SP10 protein in the sample is calculated according to the relation between the intensity of the Raman characteristic peak and the concentration of the SP10 protein; on the contrary, if the detection line does not have the characteristic peak of nai erlan a, it indicates that the sample does not contain the target SP10 protein.
Example 2
A sperm SP10 protein quantitative detection test strip based on fluorescence comprises the following steps:
A) construction of lateral flow test strips: the substrate 1 is made of a PDMS plate, the length of the PDMS plate is 198mm, and the width of the PDMS plate is 6 mm; on the substrate, a sample pad 2, a label pad 3, a chromatographic carrier 4 and a water-absorbent pad 5 are arranged in this order. Wherein the sample pad is made of a nylon membrane, the length of the sample pad is 40mm, and the width of the sample pad is 6 mm; the marking pad is made of non-woven fabric, the length of the marking pad is 10mm, and the width of the marking pad is 6 mm; the chromatography membrane is made of cellulose acetate membrane, and has a length of 80mm and a width of 6 mm; the absorbent pad is made of filter paper, and has a length of 80mm and a width of 6 mm. The detection area 6 is a single detection point consisting of aptamers corresponding to SP10 protein; the quality control region 7 is a single quality control point composed using an aptamer corresponding to the SP10 aptamer. The quality control point is arranged on the right side of the detection point. The functionalized nano material 8 is fixed on the marking pad. The sample pad and the marking pad, the marking pad and the chromatographic membrane, and the chromatographic membrane and the absorbent pad are partially overlapped, and the overlapping length is 4 mm.
B) Preparing a functionalized nano material: the nano particles with gold cores and silver shells are used as the enhancement substrate of fluorescent molecules, and the diameters of the gold cores and the sizes of the silver shells are respectively 2nm and 1.5 nm. Fluorescein isothiocyanate is connected to the surface of the gold core in a covalent connection mode, and then the silver shell is synthesized on the outer layer of the gold core; then, an aptamer corresponding to SP10 protein is modified on the surface of the gold-core silver-shell nanoparticle by means of covalent connection, and finally, a functionalized nano material is formed.
C) The use flow of the test strip for detecting the SP10 protein comprises the following steps:
firstly, a sample solution 9 containing SP10 protein is dropped on a sample pad, and the sample solution flows laterally due to capillary force. When the target SP10 protein reaches the labeling pad, the protein reacts with the functionalized nanoparticles to form an SP10 protein-nanoparticle complex; with the further flow of the liquid flow, the SP10 protein-nanoparticle complex can be combined with the aptamer on the detection point to form a sandwich structure 10; when the liquid stream reaches the control point, the functionalized nanoparticles will combine with the aptamer on the control point to form a complex 11.
And collecting signals of the detection points and the quality control points on the test strip by using a fluorescence spectrometer after 20 min.
D) And (4) analyzing results:
quality control point detection: if the fluorescence peak of fluorescein isothiocyanate appears in the fluorescence spectrum collected from the quality control point, the test strip is effective; otherwise, the test strip is invalid.
Detection point detection: if the fluorescence peak of fluorescein isothiocyanate appears in the fluorescence spectrum collected from the detection point, the SP10 protein exists in the detection sample, and at the moment, the amount of the target SP10 protein in the sample is calculated according to the relation between the intensity of the fluorescence peak and the concentration of the SP10 protein; on the contrary, if the fluorescence peak of fluorescein isothiocyanate does not appear in the detection point, the sample does not contain the target SP10 protein.
Example 3
The sperm SP10 protein quantitative detection test strip based on the magnetic signal comprises the following steps:
A) construction of lateral flow test strips: the substrate 1 is made of a PS plate, the length of the PS plate is 139mm, and the width of the PS plate is 4 mm; a sample pad 2, a marking pad 3, a chromatographic membrane 4 and a water absorption pad 5 are sequentially arranged on a substrate, wherein the sample pad is made of a non-woven membrane, the length of the sample pad is 20mm, and the width of the sample pad is 4 mm; the marking pad is made of a polyvinyl acetate film, the length of the marking pad is 8mm, and the width of the marking pad is 4 mm; the chromatography membrane is made of polyvinylidene fluoride, and has a length of 60mm and a width of 4 mm; the water absorption pad is made of cellulose-glass fiber composite, and has a length of 60mm and a width of 4 mm. The detection area 6 is a detection lattice consisting of ligands corresponding to SP10 protein; the quality control region 7 is a quality control lattice composed of ligands corresponding to SP10 ligands. The quality control lattice is arranged at the right side of the detection lattice. The functionalized nano material 8 is fixed on the marking pad. The sample pad and the marking pad, the marking pad and the chromatographic membrane, and the chromatographic membrane and the absorbent pad are partially overlapped, and the overlapping length is 3 mm.
B) Preparing a functionalized nano material: and selecting 100nm ferroferric oxide magnetic nanoparticles as a magnetic signal generator, modifying the ligand corresponding to SP10 protein on the surface of the ferroferric oxide magnetic nanoparticles in an electrostatic adsorption mode, and finally forming the functionalized nano material.
C) The use flow of the test strip for detecting the SP10 protein comprises the following steps:
firstly, a sample solution 9 containing SP10 protein is dropped on a sample pad, and the sample solution flows laterally due to capillary force. When the target SP10 protein reaches the labeling pad, the protein reacts with the functionalized nanoparticles to form an SP10 protein-nanoparticle complex; with the further flow of the liquid flow, the SP10 protein-nanoparticle complex can be combined with the ligand on the detection dot matrix to form a sandwich structure 10; when the liquid flow reaches the quality control lattice, the functionalized nano particles can be combined with the ligand on the quality control lattice to form a complex 11).
And secondly, after 10min, using a magnetic signal detector to collect signals of the detection dot matrix and the quality control dot matrix on the test strip.
D) And (4) analyzing results:
quality control dot matrix detection: if the magnetic signal can be collected from the quality control dot matrix, the test strip is effective; otherwise, the test strip is invalid.
Secondly, detecting a dot matrix: if the magnetic signal can be collected from the detection dot matrix, the SP10 protein exists in the detection sample, and the amount of the target SP10 protein in the sample is calculated according to the relation between the intensity of the magnetic signal and the concentration of the SP10 protein; on the contrary, if the magnetic signal is not detected by the detection dot array, the sample does not contain the target SP10 protein.
The above description is only a preferred embodiment of the present invention, and should not be taken as limiting the invention, and any local variations in the formulation and process thereof should be considered within the scope of the present invention.
Claims (9)
1. A sperm SP10 protein detection test strip is characterized in that the test strip is arranged in a lateral flow mode and comprises a substrate (1), a sample pad (2), a marking pad (3), a chromatographic membrane (4) and a water absorption pad (5) which are arranged above the substrate (1) in sequence from left to right, wherein one end of the chromatographic membrane (4) is arranged on the lower surface of the marking pad (3), and the upper surface of the other end is attached to the lower surface of the water absorption pad (5); the chromatographic membrane (4) is also provided with a detection area (6) and a quality control area (7); the label pad (3) is attached with a functionalized nano material capable of being combined with SP10 protein;
the substrate (1), the sample pad (2), the marking pad (3), the chromatographic membrane (4) and the absorbent pad (5) are overlapped in sequence in a staggered manner, and the overlapping length is 2-4 mm.
2. The sperm SP10 protein detection test strip of claim 1, wherein the substrate is an inert hard material that prevents liquid penetration, and specifically: a PVC plate, a PDMS plate or a PS plate, the length of which is 40-198mm, and the width of which is 2-6 mm;
the sample pad (2) is a glass fiber membrane, a nylon membrane, a non-woven membrane or a cellulose membrane, the length of the sample pad is 10-40mm, and the width of the sample pad is 2-6 mm;
the marking pad (3) is a glass fiber film, a non-woven fabric, a cellulose film or a polyvinyl acetate film, the length of the marking pad is 6-10mm, and the width of the marking pad is 2-6 mm;
the chromatographic membrane (4) is a nitrocellulose membrane, cellulose acetate, polyvinylidene fluoride or nylon membrane, the length of the chromatographic membrane is 20-80mm, and the width of the chromatographic membrane is 2-6 mm;
the absorbent pad (5) is cellulose membrane, absorbent paper, cellulose-glass fiber compound, filter paper, sponge or non-woven fabric, and has a length of 10-80mm and a width of 2-6 mm.
3. The sperm SP10 protein detection strip of claim 1, wherein the detection zone (6) is a straight line, a single point or an array of multiple points on the chromatographic membrane (4) drawn by a reagent binding to SP10 protein, and the reagent binding to SP10 protein is a capture antibody, aptamer or ligand of SP 10.
4. The sperm SP10 protein detection test strip of claim 1, wherein the quality control region (7) is a straight line, a single point or an array of multiple points drawn on the chromatographic membrane (4) by the reagent combined with the functionalized nano-material;
the reagent combined with the functionalized nanometer material is a capture antibody, an aptamer or a ligand corresponding to the SP10 protein detection substance, and the capture antibody, the aptamer or the ligand corresponding to the SP10 protein detection substance is specifically combined with the SP10 protein detection substance and does not have cross reaction with the SP10 protein; the quality control area (7) is positioned at the right side of the detection area (6).
5. The sperm SP10 protein detection test strip of claim 1, wherein a signal tag is modified on a functionalized nanomaterial that binds to SP10 protein, and a SP10 protein detection substance is fixed on the surface of the nanomaterial through electrostatic adsorption or covalent attachment, specifically: antibodies, aptamers or ligands, for use in the quantitative detection of SP10 protein;
or the nano material is not modified with any signal label, and a detection substance corresponding to the SP10 protein is fixed on the nano material, and specifically comprises the following components: an antibody, aptamer or ligand.
6. The sperm SP10 protein detection test strip of claim 5, wherein the nanomaterials are:
metal nanoparticles, including two types: the first type is a single-structure nanoparticle, specifically: gold, silver or copper nanoparticles having a particle size of 5-100 nm;
the second type is core-shell structured nanoparticles, which specifically comprise: silver core gold shell, gold core silver shell, gold core gold shell, silver core silver shell, silica core gold shell, silica core silver shell, gold core silica shell or silver core silica shell nanoparticles, wherein the size of the core is 2-80nm, and the thickness of the shell is 1.5-10 nm;
the magnetic nano material comprises the following specific components: ferroferric oxide, ferric oxide or manganese zinc ferrite nano-particles with the particle size of 5-100 nm;
③ the quantum dots specifically comprise: lead sulfide, indium phosphide/zinc sulfide or cadmium selenide/zinc sulfide quantum dots with the particle size of 5-100 nm;
the carbon nano material specifically comprises the following components: carbon nano-tubes or carbon nanospheres with a particle size of 5-100 nm;
7. the sperm SP10 protein detection test strip of claim 5, wherein said signal tag is:
the Raman dye molecule specifically comprises: p-aminophenol, p-mercaptobenzoic acid, methylene blue, rhodamine 6G, Nailan A, crystal violet, malachite green isothiocyanate or p-mercaptothiophenol;
② fluorescent dye molecules, specifically: cy3, Cy5, fluorescein isothiocyanate or phycoerythrin chlorophyll protein;
the signal label and the nano material are modified by electrostatic adsorption or covalent connection of the signal label to the surface of the nano material or the nuclear shell gap of the nuclear shell structure nano particle.
8. A quantitative detection method based on the sperm SP10 protein detection test strip of any one of claims 1-5, characterized by comprising the following steps:
s1, directly dripping the sample solution containing the SP10 protein on the sample pad (2) for 5-20 min;
s2, and carrying out signal acquisition analysis on the detection area (6) and the quality control area (7) to quantitatively detect the SP10 protein.
9. The quantitative detection method of the sperm SP10 protein detection test strip of claim 8, wherein the signal is a fluorescence, surface enhanced Raman scattering, or magnetic signal.
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