CN115290904A - In-situ hybridization HPV detection test strip, kit and preparation method thereof - Google Patents
In-situ hybridization HPV detection test strip, kit and preparation method thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of HPV detection, and relates to an in-situ hybridization HPV detection test strip, a kit and a preparation method thereof. The urine containing HPV virus is titrated on the detection test strip, and the color is developed by dyeing the C1 protein weak acid sodium tungstate, so that the amount of the HPV virus in the urine can be judged, whether a human body is infected with HPV and the carrying amount of the HPV virus can be calculated, the detection is convenient, and the result can be known in 10-15 min. The HPV detection kit can protect the detection test strip, prevent the detection test strip from being polluted, and is convenient to carry and use. The preparation method is simple, easy to operate, low in cost and easy for industrial production.
Description
Technical Field
The invention belongs to the technical field of HPV detection, and particularly relates to an in-situ hybridization HPV detection test strip, a kit and a preparation method thereof.
Background
HPV (human papilloma virus) is a papilloma vacuolatum virus A genus belonging to papovaviridae family, is a spherical DNA virus and can cause squamous epithelial proliferation of human skin mucosa. The HPV test mainly detects whether a human carries HPV virus or not along with the rapid increase of the incidence rate of condyloma acuminatum in venereal diseases and the increase of cervical cancer, anal cancer and the like, and whether the human is infected with HPV can be detected by a staining microscopy method, an HPV DNA detection method or a serological test.
At present, the detection method of HPV early infection is limited to smear cytomorphology examination of cervical sample and DNA detection of HPV, and sampling and detection must be carried out in professional institutions such as hospitals, so the cervical sample detection method is far not suitable for large-scale crowd self-detection.
Disclosure of Invention
The invention aims to provide an in-situ hybridization HPV detection test strip which can carry out self-detection, is convenient and simple to detect and has reliable results, so as to solve the problems in the background technology.
In order to achieve the technical purpose, the technical scheme of the invention is as follows:
an in-situ hybridization HPV detection test strip comprises a first chromatographic pad, detection test paper and a second chromatographic pad which are sequentially connected, wherein glass fibers are bonded below the second chromatographic pad, the length of the glass fibers is greater than that of the second chromatographic pad, and absorbent cotton is bonded below one end, close to the detection test paper, of the glass fibers; the detection test paper is obtained by soaking test paper in a reagent solution and then drying the soaked test paper, wherein the reagent solution comprises 5-10% of HPVC1 protein in-situ hybridization solution, 5% of citric acid, 10-20% of sodium acetate, 10-40% of sodium tungstate, 5% of mercury chloride and the balance of water by weight, and the HPVC1 protein in-situ hybridization solution is formed by in-situ hybridization of HPVC1 protein and a cysteine solution or a cysteine hydrochloride solution.
The test paper used before soaking is generally qualitative filter paper. In the reagent solution, citric acid reacts with sodium tungstate to form weak acid sodium tungstate, sodium acetate strengthens the fixation of tungsten ions to the weak sodium tungstate, and mercuric chloride can maintain the stability of the reagent solution in the test strip and eliminate negative interference elements.
Preferably, the weight ratio of the HPVC1 protein to the cysteine solution or the cysteine hydrochloride solution is 1-10.
The invention also aims to provide an in-situ hybridization HPV detection kit, which comprises a shell, wherein the in-situ hybridization HPV detection test strip is arranged in the shell, the shell is provided with a urine hole and an observation hole, the observation hole is positioned above the detection test strip, and the urine hole is positioned above the glass fiber.
As an improvement, the casing includes casing and lower casing, go up the casing with the casing is pegged graft down, it is protruding to be equipped with first hole post, second in the upper casing observe the hole urine hole and imbibition cotton fixed slot, the both sides diagonal angle of observing the hole is equipped with third hole post and fourth hole post, it is protruding that internal first arch, second hole post, third and the fourth of being equipped with of inferior valve is protruding, first arch with first hole post looks adaptation, the second arch with second hole post looks adaptation, the third arch with third hole post looks adaptation, the fourth arch with fourth hole post looks adaptation, first chromatography pad with the second is protruding to be connected, it locates to detect the paper card third hole post with between the fourth hole post, imbibition cotton card is located in the imbibition cotton fixed slot.
Preferably, the second protrusion is a plastic staple.
As a further improvement, a groove is arranged in the lower shell body and close to the first chromatographic pad, and a drying sheet is arranged in the groove. The drying tablets are preferably silica gel mineral drying tablets.
Preferably, the shape of the housing is a strip or a leaf.
As an improvement, the shell is provided with a color comparison card.
The invention also aims to provide a preparation method of the in-situ hybridization HPV detection test strip, which comprises the following steps:
(1) Taking C1 protein human papilloma virus in urine to carry out PCR-DNA nucleic acid multiplication to obtain HPVC1 protein;
(2) Adding purified water into cysteine or cysteine hydrochloride to prepare a cysteine solution or cysteine hydrochloride solution with the concentration of 24 mmol/L;
(3) Mixing the HPVC1 protein with the cysteine solution or cysteine hydrochloride solution, and standing for 48 hours to obtain an HPVC1 protein in-situ hybridization solution;
(4) Taking citric acid, sodium tungstate and sodium acetate according to a proportion, uniformly mixing, and standing to obtain a mixed solution;
(5) Taking the HPVC1 protein in-situ hybridization solution, mercuric chloride, the mixed solution and water according to a certain proportion, uniformly mixing, standing for 2 hours to obtain a reagent solution, putting the test paper into the reagent solution, soaking for 10min, taking out, and drying to obtain test paper;
(6) And respectively bonding a first chromatography pad and a second chromatography pad to two ends of the detection test paper, bonding glass fibers below the second chromatography pad, and bonding liquid absorption cotton to one end, close to the detection test paper, of the glass fibers.
In the step (1), the urine C1 protein refers to the HPV virus coat protein C1, also called HPV L1 protein, and refers to the HPV virus coat. Actively infected HPV virus mainly comprises DNA containing virus and coat protein C1, while latent HPV virus only has DNA without coat protein C1, so that detection of HPV virus mainly shows expression of coat protein C1 to determine whether HPV is infected or not.
For latent HPV infection, HPV virus DNA is integrated with host DNA in a mosaic mode, so that active infection of HPV virus is converted into negative, the virus load is reduced, the titer of latent infection virus is changed, the infection degree of the HPV is qualitatively shown through color development, the coat protein C1 also has an antigen epitope, and a protein receptor is arranged on the surface of a host cell, so that the infection gradient of the HPV is judged.
The invention also aims to provide a preparation method of the in situ hybridization HPV detection kit, which comprises the following steps: and cutting the in-situ hybridization HPV detection test strip according to the size and the shape of the shell, and filling the cut test strip into the shell to obtain the test strip.
Due to the adoption of the technical scheme, the invention has the beneficial effects that:
the HPV detection test strip provided by the invention is obtained by soaking a solution prepared by carrying out in-situ hybridization on C1 protein-containing human papilloma virus carried in urine and cysteine, and then carrying out cross reaction with citric acid, sodium tungstate and sodium acetate, titrating the urine containing the C1 protein-containing human papilloma virus on the HPV detection test strip, and dyeing and developing through C1 protein weak acid sodium tungstate, so that the amount of HPV virus in the urine can be judged, and further whether a human body is infected with HPV and the carrying amount can be calculated. If the urine HPV is detected to be positive, the detection test strip is dyed to be blue, the darker the color is, the more HPV virus carried in the urine is shown, and the lighter the dyeing is, the less HPV virus carried in the urine is shown; if the urine HPV detection is negative, the detection test strip shows no color (colorless), which indicates that the urine is not infected with HPV virus. The HPV detection kit can protect the detection test strip, prevent the detection test strip from being polluted, and is convenient to carry and use. The preparation method provided by the invention is simple, easy to operate, low in cost and easy for industrial production.
Drawings
FIG. 1 is a front view of an in situ hybridization HPV detection test strip provided by the present invention;
FIG. 2 is a side view of the in situ hybridization HPV detection test strip provided by the present invention;
FIG. 3 is a front view of the in situ hybridization HPV detection kit provided in example 5;
FIG. 4 is a schematic structural view of an upper case provided in embodiment 5;
FIG. 5 is a schematic structural view of a lower case provided in embodiment 5;
FIG. 6 is a front view of the in situ hybridization HPV detection kit provided in example 6;
FIG. 7 is a schematic structural view of an upper case provided in embodiment 6;
FIG. 8 is a schematic structural view of a lower case provided in embodiment 6;
FIG. 9 is a front view of the in situ hybridization HPV detection kit provided in example 7;
FIG. 10 is a rear view of the in situ hybridization HPV detection kit provided in example 7;
FIG. 11 is a schematic structural view of an upper case provided in embodiment 7;
FIG. 12 is a schematic structural view of a lower casing provided in embodiment 7;
FIG. 13 is a graph showing the results of an accuracy test of the detection kit provided in example 5;
FIG. 14 is a graph showing the results of an accuracy test of the detection kit provided in example 6;
FIG. 15 is a graph showing the results of a specificity test of the detection kit provided in example 5;
FIG. 16 is a graph showing the results of a repetitive experiment using the detection kit provided in example 6;
wherein: 1-shell, 2-test paper, 3-first chromatography pad, 4-second chromatography pad, 5-glass fiber, 6-absorbent cotton, 7-dry sheet, 8-color chart, 9-support edge, 10-arc bulge, 11-upper shell, 12-lower shell, 111-first pore column, 112-second bulge, 113-observation pore, 114-urine pore, 115-absorbent cotton fixing groove, 116-third pore column, 117-fourth pore column, 118-color comparison pore, 121-first bulge, 122-second pore column, 123-third bulge, 124-fourth bulge and 125-groove.
Detailed Description
The present invention will be further described with reference to the following detailed description and accompanying drawings. Wherein the showings are for the purpose of illustration only and not for the purpose of limiting the same, the same is shown by way of illustration only and not in the form of limitation; to better illustrate the embodiments of the present invention, some parts of the drawings may be omitted, enlarged or reduced, and do not represent the size of an actual product; it will be understood by those skilled in the art that certain well-known structures in the drawings and descriptions thereof may be omitted.
The same or similar reference numerals in the drawings of the embodiments of the present invention correspond to the same or similar components; in the description of the present invention, it should be understood that if there is an orientation or positional relationship indicated by the terms "upper", "lower", "left", "right", etc., based on the orientation or positional relationship shown in the drawings, it is only for convenience of description and simplification of the description, but it is not intended to indicate or imply that the device or element referred to must have a specific orientation, be constructed in a specific orientation and operate, and therefore the terms describing the positional relationship in the drawings are only used for illustrative purposes and are not to be construed as limiting the present patent, and it is possible for one of ordinary skill in the art to understand the specific meaning of the above terms according to the specific situation.
Example 1
Preparing HPVC1 protein in situ hybridization solution: taking C1 protein human papilloma virus in urine, carrying out PCR-DNA nucleic acid multiplication to enable the C1 protein to be inlaid in DNA cells, and obtaining HPVC1 protein; adding purified water into cysteine to prepare a cysteine solution with the concentration of 24 mmol/L; mixing the HPVC1 protein and the cysteine solution, wherein the weight ratio of the HPVC1 protein to the cysteine solution is 1:99, standing for 48 hours to obtain the HPVC1 protein in situ hybridization solution.
The reagent liquid formula comprises: 5% of HPVC1 protein in situ hybridization solution, 5% of citric acid, 15% of sodium acetate, 20% of sodium tungstate, 5% of mercury chloride and the balance of water.
Taking the citric acid, the sodium tungstate and the sodium acetate according to the proportion, uniformly mixing, and standing to obtain a mixed solution; and then taking the HPVC1 protein in-situ hybridization solution, mercuric chloride and water according to a certain proportion, uniformly mixing the mixed solution, standing for 2 hours to obtain a reagent solution, putting the test paper into the reagent solution, soaking for 10min, taking out, and drying at 50-75 ℃ to obtain the test paper.
Example 2
Preparing HPVC1 protein in situ hybridization solution: taking C1 protein human papilloma virus in urine, carrying out PCR-DNA nucleic acid multiplication to enable the C1 protein to be inlaid in DNA cells, and obtaining HPVC1 protein; adding purified water into cysteine to prepare a cysteine solution with the concentration of 24 mmol/L; mixing the HPVC1 protein and the cysteine solution, wherein the weight ratio of the HPVC1 protein to the cysteine solution is 5: and 95, standing for 48 hours to obtain the HPVC1 protein in-situ hybridization solution.
The reagent liquid formula comprises: 8% of HPVC1 protein in situ hybridization solution, 5% of citric acid, 10% of sodium acetate, 40% of sodium tungstate, 5% of mercury chloride and the balance of water.
Taking the citric acid, the sodium tungstate and the sodium acetate according to the proportion, uniformly mixing, and standing to obtain a mixed solution; and then taking the HPVC1 protein in-situ hybridization solution, mercuric chloride and water according to a certain proportion, uniformly mixing the mixed solution, standing for 2 hours to obtain a reagent solution, putting the test paper into the reagent solution, soaking for 10min, taking out, and drying at 50-75 ℃ to obtain the test paper.
Example 3
Preparing HPVC1 protein in situ hybridization solution: taking C1 protein human papilloma virus in urine to carry out PCR-DNA nucleic acid multiplication so as to enable the C1 protein to be inlaid in DNA cells, thus obtaining HPVC1 protein; adding purified water into cysteine hydrochloride to prepare cysteine hydrochloride solution with the concentration of 24 mmol/L; mixing the HPVC1 protein and the cysteine hydrochloride solution, wherein the weight ratio of the HPVC1 protein to the cysteine hydrochloride solution is 10: standing for 48 hours to obtain the HPVC1 protein in situ hybridization solution.
The reagent liquid formula comprises: 10% of HPVC1 protein in situ hybridization solution, 5% of citric acid, 20% of sodium acetate, 10% of sodium tungstate, 5% of mercury chloride and the balance of water.
Taking the citric acid, the sodium tungstate and the sodium acetate according to the proportion, uniformly mixing, and standing to obtain a mixed solution; and then taking the HPVC1 protein in-situ hybridization solution, mercuric chloride and water according to a certain proportion, uniformly mixing the mixed solution, standing for 2 hours to obtain a reagent solution, putting the test paper into the reagent solution, soaking for 10min, taking out, and drying at 50-75 ℃ to obtain the test paper.
Example 4
Preparing HPVC1 protein in situ hybridization solution: taking C1 protein human papilloma virus in urine, carrying out PCR-DNA nucleic acid multiplication to enable the C1 protein to be inlaid in DNA cells, and obtaining HPVC1 protein; adding purified water into cysteine hydrochloride to prepare cysteine hydrochloride solution with the concentration of 24 mmol/L; mixing the HPVC1 protein and the cysteine hydrochloride solution, wherein the weight ratio of the HPVC1 protein to the cysteine hydrochloride solution is 7:93, standing for 48 hours to obtain the HPVC1 protein in-situ hybridization solution.
The reagent liquid formula comprises: 7% of HPVC1 protein in situ hybridization solution, 5% of citric acid, 12% of sodium acetate, 30% of sodium tungstate, 5% of mercury chloride and the balance of water.
Taking the citric acid, the sodium tungstate and the sodium acetate according to the proportion, uniformly mixing, and standing to obtain a mixed solution; and then taking the HPVC1 protein in-situ hybridization solution, mercuric chloride and water according to a certain proportion, uniformly mixing the mixed solution, standing for 2 hours to obtain a reagent solution, putting the test paper into the reagent solution, soaking for 10min, taking out, and drying at 50-75 ℃ to obtain the test paper.
Example 5
As shown in fig. 1-2, an in situ hybridization HPV detection test strip comprises a detection test paper 2, a first chromatographic pad 3, a second chromatographic pad 4, glass fibers 5 and absorbent cotton 6 prepared in embodiments 1-4, wherein the first chromatographic pad 3 is adhered to the left end of the detection test paper 2, the second chromatographic pad 4 is adhered to the right end of the detection test paper 2, the ends of the first chromatographic pad 3 and the second chromatographic pad 4, which are adhered to the detection test paper 2, are located on the detection test paper 2, the glass fibers 5 are adhered to the lower side of the second chromatographic pad 4, the length of the glass fibers 5 is greater than that of the second chromatographic pad 4, the absorbent cotton 6 is adhered to the lower side of the glass fibers 5, and the absorbent cotton 6 is adhered to the end of the glass fibers 5, which is close to the detection test paper 2.
During detection, the urine is dropped on the glass fiber 5, the urine flows towards the second chromatographic pad 4 through the glass fiber 5, the second chromatographic pad 4 can filter out some impurities in the urine, when the urine flows to the liquid absorption cotton 6, the flow speed of the urine can be controlled through the size of the liquid absorption cotton 6, so that the color development time of the test paper 2 can be controlled, if the liquid absorption cotton 6 is large, the flow speed of the urine can be accelerated, the color development time of the test paper 2 is slightly faster, on the contrary, if the liquid absorption cotton 6 is small, the flow speed of the urine is relatively slow, the color development time of the test paper 2 is slightly slower, the first chromatographic pad 3 can block the urine from continuously flowing, so that the display time of the test paper 2 is relatively stable, and the phenomenon that the urine continuously flows to cause the color development of the test paper 2 to be unstable is avoided.
As shown in fig. 3-5, an in situ hybridization HPV detection kit includes a housing 1, the housing 1 is in a strip shape, the housing 1 includes an upper housing 11 and a lower housing 12, the edges of the upper housing 11 and the lower housing 12 are connected and fixed by means of insertion of a protrusion and a hole column, a first hole column 111, a second protrusion 112, an observation hole 113, a urine hole 114 and a fixing groove 115 for absorbent cotton are provided in the upper housing 11, a third hole column 116 and a fourth hole column 117 are symmetrically provided at two corners of the observation hole 113, the detection test paper 2 manufactured in embodiments 1-4 is clamped between the third hole column 116 and the fourth hole column 117, a first chromatography pad 3 and a second chromatography pad 4 are respectively adhered to two ends of the detection test paper 2, a through hole is provided on the first chromatography pad 3, the through hole can penetrate through the second protrusion 112 and hang on the second protrusion 112, the second protrusion 112 is preferably plastic, and this design can prevent the first chromatography pad 3 from falling off from the plastic clamping nail, and then the second protrusion 112 is inserted into the second hole column 122, so as to fix the first chromatography pad 3. The sight glass 113 is located above the test strip 2 and the urine glass 114 is located above the glass fibre 5. The below of second chromatography pad 4 bonds and has glass fiber 5, and glass fiber 5 bonds and has the imbibition cotton 6 near the one end below of test paper strip 2, thereby imbibition cotton 6 impresses in imbibition cotton fixed slot 115 can fix second chromatography pad 4 and glass fiber 5 in last casing 11, and the inside wall of imbibition cotton fixed slot 115 is equipped with the protruding 10 of a plurality of arcs, and the protruding 10 of arc can further fix imbibition cotton 6 in imbibition cotton fixed slot 115.
A first protrusion 121, a second hole column 122, a third protrusion 123 and a fourth protrusion 124 are arranged in the lower shell 12, a groove 125 is arranged in the lower shell 12 at a position close to the first chromatographic pad 3, and a drying sheet 7 is arranged in the groove 125. The main component of the drying sheet 7 is silica gel mineral powder. The drying sheet 7 can keep the test paper 2 dry, and prevent the test paper from absorbing moisture to affect the detection result. First arch 121 and first hole post 111 looks adaptation, second arch 112 and second hole post 122 looks adaptation, third arch 123 and third hole post 116 looks adaptation, fourth arch 124 and fourth hole post 117 looks adaptation, through inserting first arch 121, second arch 112, third arch 123 and fourth arch 124 respectively in first hole post 111, second hole post 122, third hole post 116 and fourth hole post 117, can be with upper casing 11 and lower casing 12 closed formation casing 1, and will inhale liquid detection test paper 2, first chromatography pad 3, second chromatography pad 4, glass fiber 5, cotton 6 and dry sheet 7 fix in casing 1, avoid it to remove at will.
The preparation method of the in-situ hybridization HPV detection kit comprises the following steps: the detection test paper 2, the first chromatographic pad 3, the second chromatographic pad 4, the glass fiber 5 and the absorbent cotton 6 are firstly cut into required shapes and sizes according to the shape of the shell 1, the first chromatographic pad 3 and the second chromatographic pad 4 are respectively bonded at two ends of the detection test paper 2, the glass fiber 5 is bonded with the second chromatographic pad 4, the absorbent cotton 6 is bonded with one end of the glass fiber 5, the first chromatographic pad 3 is hung on the second protrusion 112, the detection test paper 2 is clamped between the third pore column 116 and the fourth pore column 117, the absorbent cotton 6 is pressed into the absorbent cotton fixing groove 115, so that the second chromatographic pad 4, the glass fiber 5 and the absorbent cotton 6 can be fixed in the upper shell 11, the drying sheet 7 is placed in the groove 125, and the first protrusion 121, the second protrusion 112, the third protrusion 123 and the fourth protrusion 124 are respectively inserted into the first pore column 111, the second pore column 122, the third pore column 116 and the fourth pore column 117, so that the upper shell 11 and the inserting body 12 are combined to obtain the plug-in-shell.
Example 6
As shown in FIGS. 6 to 8, the in situ hybridization HPV detection kit of the present example is substantially the same in structure as the in situ hybridization HPV detection kit of example 5, except that: the shape of the housing 1 of this embodiment is a leaf shape, the upper housing 11 is provided with a colorimetric hole 118, the colorimetric card 8 is provided below the colorimetric hole 118, the colorimetric card 8 is provided with a color developing block No. 1-9, and the colorimetric card 8 is fixed in the upper housing 11. The user can judge the negative or positive of urine and the amount of HPV virus by comparing the detected color with the color developing block on the color comparison card 8.
Example 7
As shown in fig. 9 to 12, the in situ hybridization HPV detection kit of the present embodiment has substantially the same structure as the in situ hybridization HPV detection kit of example 5, except that: the urine hole 114 is not arranged in the embodiment, the glass fiber 5 extends out of the shell 1, and the urine can be directly sprayed on the exposed glass fiber 5 for detection during use.
In embodiments 5 to 7, a plurality of supporting ribs 9 are further disposed in the upper housing 11 and the lower housing 12, the supporting ribs 9 can prevent the housing 1 from deforming, so as to increase the stability and pressure resistance, and the supporting ribs 9 can also compress the test paper 2, the first chromatographic pad 3, the second chromatographic pad 4, the glass fiber 5, and the absorbent cotton 6 in the housing 1 to a certain extent, so as to prevent the test paper from shaking or shifting in the housing 1.
Accuracy test
Detection of cysteine aqueous solutions at various concentration levels was performed with the in situ hybridization HPV detection kit of example 5, reference concentrations of cysteine solutions: 0.7mmol/L, 1.4mmol/L, 2.8mmol/L, 4mmol/L, 12mmol/L, 16mmol/L, 24mmol/L. Each concentration level was repeated 3 times, comparing the difference in magnitude of the detection result and the reference solution's labeled concentration, with a relative deviation of the detection result from the labeled concentration of no more than 90%. The detection result is shown in FIG. 13. As can be seen from FIG. 13, the color development result of the in situ hybridization HPV detection kit of example 5 is compared with that of a handheld color card, and the higher the concentration of the cysteine aqueous solution is, the darker the color development is, which indicates that the detection result of the in situ hybridization HPV detection kit is accurate.
The in situ hybridization HPV detection kit of example 6 was used to detect cysteine aqueous solutions at various concentration levels, and the reference concentrations of the cysteine aqueous solutions were: 0.75mmol/L, 1.5mmol/L, 6mmol/L, 12mmol/L. Each concentration level was repeated 3 times, comparing the difference in magnitude of the detection result and the reference solution's labeled concentration, with a relative deviation of the detection result from the labeled concentration of no more than 90%. The detection result is shown in FIG. 14. As can be seen from FIG. 14, the color development result of the in situ hybridization HPV detection kit of example 6 is compared with that of a handheld color development card, and the greater the concentration of the cysteine aqueous solution is, the darker the color development is, indicating that the detection result of the in situ hybridization HPV detection kit is accurate.
Experiment of specificity
The same negative samples were each tested with the detection kit of example 5 and had cysteine concentrations of 0.6mmol/L, 1.2mmol/L, 2.4mmol/L, 4mmol/L, 8mmol/L, 12mmol/L, 16mmol/L and 24mmol/L, respectively. The measurement is repeated 3 times at each concentration level, and the consistency degree of the detection results of the detection items is observed and compared, the results are negative, and the relative deviation is not more than 90%. The detection result is shown in FIG. 15. As can be seen from FIG. 15, the specificity of the present invention is strong.
Repeatability test
The cysteine solution with the reference concentration of 24mmol/L is repeatedly measured by the detection kit in the embodiment 6 for 4 times, laid flat and kept still for 10min, the difference of the detection result and the magnitude of the labeled concentration of the reference solution is compared, and the relative deviation of the detection result and the labeled concentration is not more than 90%. The detection result is shown in FIG. 16. As can be seen from fig. 16, the present invention is good in reproducibility.
The use method of the detection kit provided by the invention comprises the following steps: taking the initial 5-10ml urine when urinating for the first time from morning, dripping the urine into the urine hole, keeping flat and standing for 10-15 minutes to wait for a detection result, and comparing the color displayed in the observation hole with the color in the color comparison card, thereby determining whether the urine is infected with HPV and the carrying amount, wherein the normal urine does not contain HPV, and the detection test paper is colorless and shows negative. The urine containing HPV shows positive, the detection test paper shows color, and the darker the color shows that the HPV carrying amount is larger.
The above-described embodiments of the present invention should not be construed as limiting the scope of the present invention. Any other corresponding changes and modifications made according to the technical idea of the present invention should be included in the protection scope of the claims of the present invention.
Claims (10)
1. The in-situ hybridization HPV detection test strip is characterized by comprising a first chromatographic pad, detection test paper and a second chromatographic pad which are sequentially connected, wherein glass fibers are bonded below the second chromatographic pad, the length of the glass fibers is greater than that of the second chromatographic pad, and absorbent cotton is bonded below one end, close to the detection test paper, of the glass fibers; the detection test paper is obtained by soaking test paper in a reagent solution and then drying the soaked test paper, wherein the reagent solution comprises 5-10% of HPVC1 protein in-situ hybridization solution, 5% of citric acid, 10-20% of sodium acetate, 10-40% of sodium tungstate, 5% of mercury chloride and the balance of water by weight, and the HPVC1 protein in-situ hybridization solution is formed by in-situ hybridization of HPVC1 protein and a cysteine solution or a cysteine hydrochloride solution.
2. The in situ hybridization HPV detection test strip according to claim 1, characterized in that the weight ratio of the HPVC1 protein to the cysteine solution or the cysteine hydrochloride solution is 1-10.
3. An in-situ hybridization HPV detection kit, which comprises a shell, and is characterized in that the in-situ hybridization HPV detection test strip of claim 1 is arranged in the shell, a urine hole and an observation hole are arranged on the shell, the observation hole is positioned above the detection test strip, and the urine hole is positioned above the glass fiber.
4. The in situ hybridization HPV detection kit according to claim 3 wherein the housing comprises an upper housing and a lower housing, the upper housing is plugged with the lower housing, the upper housing is internally provided with a first hole column, a second protrusion, the observation hole, the urine hole and a liquid-absorbing cotton fixing groove, the observation hole is diagonally provided with a third hole column and a fourth hole column at both sides, the lower housing is internally provided with a first protrusion, a second hole column, a third protrusion and a fourth protrusion, the first protrusion is adapted to the first hole column, the second protrusion is adapted to the second hole column, the third protrusion is adapted to the third hole column, the fourth protrusion is adapted to the fourth hole column, the first chromatographic pad is connected to the second protrusion, the detection test paper card is arranged between the third hole column and the fourth hole column, and the liquid-absorbing cotton card is arranged in the liquid-absorbing cotton fixing groove.
5. The in situ hybridization HPV detection kit of claim 4, in which the second protrusion is a plastic staple.
6. The in situ hybridization HPV detection kit according to claim 4 wherein the lower housing is provided with a recess in a location adjacent to the first chromatographic pad, the recess being provided with a drying sheet therein.
7. The in situ hybridization HPV detection kit of claim 3 wherein the housing is in the shape of a strip or leaf.
8. The in situ hybridization HPV detection kit according to claim 3 characterised in that a colour chart is provided on the housing.
9. The method for preparing the in-situ hybridization HPV detection test strip of claim 1, which is characterized by comprising the following steps:
(1) Taking C1 protein human papilloma virus in urine to carry out PCR-DNA nucleic acid multiplication to obtain HPVC1 protein;
(2) Adding purified water into cysteine or cysteine hydrochloride to prepare a cysteine solution or cysteine hydrochloride solution with the concentration of 24 mmol/L;
(3) Mixing the HPVC1 protein with the cysteine solution or cysteine hydrochloride solution, and standing for 48 hours to obtain an HPVC1 protein in-situ hybridization solution;
(4) Taking citric acid, sodium tungstate and sodium acetate according to a proportion, uniformly mixing, and standing to obtain a mixed solution;
(5) Taking the HPVC1 protein in-situ hybridization solution, mercuric chloride, the mixed solution and water according to a certain proportion, uniformly mixing, standing for 2 hours to obtain a reagent solution, putting the test paper into the reagent solution, soaking for 10min, taking out, and drying to obtain test paper;
(6) And respectively bonding a first chromatography pad and a second chromatography pad to two ends of the detection test paper, bonding glass fibers below the second chromatography pad, and bonding liquid absorption cotton to one end, close to the detection test paper, of the glass fibers.
10. A method for preparing the in situ hybridization HPV detection kit of claim 3, characterized by comprising the following steps: the in situ hybridization HPV detection test strip of claim 1 is cut according to the size and shape of the shell, and is filled into the shell after being cut.
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Cited By (1)
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