CN220323330U - Device for rapidly detecting cathepsin Z - Google Patents
Device for rapidly detecting cathepsin Z Download PDFInfo
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- CN220323330U CN220323330U CN202321926118.8U CN202321926118U CN220323330U CN 220323330 U CN220323330 U CN 220323330U CN 202321926118 U CN202321926118 U CN 202321926118U CN 220323330 U CN220323330 U CN 220323330U
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- cathepsin
- quality control
- control line
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- 102000011937 Cathepsin Z Human genes 0.000 title claims abstract description 38
- 108010061117 Cathepsin Z Proteins 0.000 title claims abstract description 38
- 238000001514 detection method Methods 0.000 claims abstract description 60
- 238000003908 quality control method Methods 0.000 claims abstract description 23
- 239000000020 Nitrocellulose Substances 0.000 claims abstract description 18
- 239000007788 liquid Substances 0.000 claims abstract description 18
- 239000012528 membrane Substances 0.000 claims abstract description 18
- 229920001220 nitrocellulos Polymers 0.000 claims abstract description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 11
- 238000010521 absorption reaction Methods 0.000 claims abstract description 9
- 238000004587 chromatography analysis Methods 0.000 claims abstract description 8
- 239000003550 marker Substances 0.000 claims abstract description 8
- 241000283707 Capra Species 0.000 claims abstract description 5
- 230000000694 effects Effects 0.000 claims abstract description 5
- 241001529936 Murinae Species 0.000 claims abstract description 4
- 102100026657 Cathepsin Z Human genes 0.000 claims abstract 3
- 101000910979 Homo sapiens Cathepsin Z Proteins 0.000 claims abstract 3
- 239000004005 microsphere Substances 0.000 claims description 17
- 230000002745 absorbent Effects 0.000 claims description 10
- 239000002250 absorbent Substances 0.000 claims description 10
- 239000000463 material Substances 0.000 claims description 5
- 229920006267 polyester film Polymers 0.000 claims description 5
- 239000012491 analyte Substances 0.000 claims description 3
- 229910052747 lanthanoid Inorganic materials 0.000 claims description 3
- 150000002602 lanthanoids Chemical class 0.000 claims description 3
- 150000001875 compounds Chemical class 0.000 claims 1
- 238000005507 spraying Methods 0.000 abstract description 5
- 238000002965 ELISA Methods 0.000 abstract description 4
- 238000006243 chemical reaction Methods 0.000 abstract description 3
- 230000035484 reaction time Effects 0.000 abstract description 3
- 238000000034 method Methods 0.000 abstract description 2
- 230000008569 process Effects 0.000 abstract description 2
- 239000000523 sample Substances 0.000 description 27
- 239000000243 solution Substances 0.000 description 9
- 229910052751 metal Inorganic materials 0.000 description 3
- 239000002184 metal Substances 0.000 description 3
- 239000007987 MES buffer Substances 0.000 description 2
- 241001494479 Pecora Species 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 230000002776 aggregation Effects 0.000 description 2
- 238000004220 aggregation Methods 0.000 description 2
- 238000003317 immunochromatography Methods 0.000 description 2
- 201000011461 pre-eclampsia Diseases 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- 102000005927 Cysteine Proteases Human genes 0.000 description 1
- 108010005843 Cysteine Proteases Proteins 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000002759 chromosomal effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000002288 golgi apparatus Anatomy 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 238000006748 scratching Methods 0.000 description 1
- 230000002393 scratching effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
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- Investigating Or Analysing Biological Materials (AREA)
Abstract
The utility model provides a device for rapidly detecting cathepsin Z, which relates to the technical field of gynecological detection and comprises the following components: the detection part comprises a bottom plate, wherein the bottom plate is sequentially lapped and stuck with a sample pad, a bonding pad, a nitrocellulose membrane and a water absorption pad from left to right, so that sample liquid dropped on the sample pad sequentially passes through the bonding pad and the nitrocellulose membrane to swim towards the water absorption pad; spraying CTSZ antibody on the bonding pad; the display part comprises a detection line and a quality control line; the detection line and the quality control line are arranged on the nitrocellulose membrane along the flowing direction of the sample liquid, and the detection line is an anti-CTSZ antibody coated with a murine source and is used for forming a marker when the complex of the to-be-detected object moves forwards under the chromatography effect; the quality control line is goat anti-mouse IgG, and is used for forming a marker when an unbound object to be detected continuously moves forward, so that the problems of long reaction time, complex reaction operation, need of matching with a high-precision instrument and low efficiency of the whole detection process of ELISA detection are solved.
Description
Technical Field
The utility model relates to the technical field of gynecological detection, in particular to a device for rapidly detecting cathepsin Z.
Background
Cathepsin Z (CTSZ), a novel human cysteine protease, has a short propeptide domain and unique chromosomal location, and its function is related to vesicle-mediated and transport to the golgi apparatus and subsequent modification; long-term researches show that the content of cathepsin Z in peripheral blood of preeclampsia pregnant women is obviously higher than that of healthy pregnant women, so that based on the research result, the cathepsin Z is used as a detection index of preeclampsia to be applied to detection products;
however, the detection mode of cathepsin Z in hospitals is mainly single detection, and samples are detected by ELISA detection, so that ELISA detection has long reaction time, complex reaction operation, high-precision instrument matching and lower efficiency of the whole detection process.
Disclosure of Invention
The utility model provides a device for rapidly detecting cathepsin Z, which specifically comprises:
the detection part comprises a bottom plate, wherein the bottom plate is sequentially lapped and stuck with a sample pad, a bonding pad, a nitrocellulose membrane and a water absorption pad from left to right, so that sample liquid dropped on the sample pad sequentially passes through the bonding pad and the nitrocellulose membrane to swim towards the water absorption pad; the CTSZ antibody is sprayed on the binding pad and used for being combined with a part of the to-be-detected objects in the sample liquid to form to-be-detected object complexes;
the display part comprises a detection line and a quality control line; the detection line and the quality control line are arranged on the nitrocellulose membrane along the flowing direction of the sample liquid, and the detection line is an anti-CTSZ antibody coated with a murine source and is used for forming a marker when the complex of the to-be-detected object moves forwards under the chromatography effect; the quality control line is goat anti-mouse IgG and is used for forming a marker when an unbound analyte continues to advance.
Optionally, the detector comprises a protection part sleeved on the outer side of the detection part; the protection part comprises a box body; the middle part of the upper end of the box body is provided with a round-corner rectangular observation window; the observation window is positioned at the upper end of the nitrocellulose membrane, and the display part is displayed through the observation window.
Optionally, a conical placing window is formed on the left side of the upper end of the box body; the placing window is positioned at the upper end of the sample pad and is used for dripping sample liquid into the sample pad.
Optionally, the bonding pad is formed from a polyester film.
Optionally, the surface of the bonding pad is sprayed with a CTSZ antibody material layer, and lanthanide fluorescent microspheres are coupled on the CTSZ antibody material layer.
Optionally, the diameter of the fluorescent microsphere is 120mm.
Optionally, the interval between the quality control line and the detection line is set to be 5mm-8mm.
Optionally, the widths of the quality control line and the detection line are set to be 3mm-5mm.
Optionally, the chromatography is directed from the conjugate pad to the absorbent pad.
Optionally, the absorbent pad is absorbent paper.
Advantageous effects
According to the utility model, the cathepsin Z is detected by a fluorescence immunochromatography detection technology, the reaction time is greatly shortened to 15 minutes compared with 2-4 hours of ELISA, the operation is simple, a sample and a buffer solution are fully mixed and then are dripped into a detection area, the detection value can be read by a hand-held fluorescence detector, and the detection efficiency is greatly improved.
Drawings
In order to more clearly illustrate the technical solution of the embodiments of the present utility model, the drawings of the embodiments will be briefly described below.
The drawings described below are only for illustration of some embodiments of the utility model and are not intended to limit the utility model.
In the drawings:
fig. 1 is a schematic perspective view showing the whole structure of a cathepsin Z detection device according to an embodiment of the present utility model.
FIG. 2 shows a schematic perspective cross-sectional view of the whole of a cathepsin Z detection device according to an embodiment of the utility model.
Fig. 3 is a schematic perspective view showing a detecting section of a cathepsin Z detecting device according to an embodiment of the present utility model.
FIG. 4 shows an enlarged schematic view of a portion of FIG. 3 of a cathepsin Z detection device according to an embodiment of the utility model.
FIG. 5 shows a cross-sectional view of the entirety of a cathepsin Z detection device according to an embodiment of the utility model.
List of primary reference numerals
1. A protection part; 101. a case body; 102. an observation window; 103. placing a window;
2. a detection unit; 201. a bottom plate; 202. a sample pad; 203. a bonding pad; 204. a nitrocellulose membrane; 205. a water absorbing pad;
3. a display unit; 301. a detection line; 302. and a quality control line.
Detailed Description
In order to make the objects, aspects and advantages of the technical solution of the present utility model more clear, the technical solution of the embodiment of the present utility model will be clearly and completely described below with reference to the accompanying drawings of the specific embodiment of the present utility model.
Examples: please refer to fig. 1 to 5:
the utility model provides a device for rapidly detecting cathepsin Z, which comprises:
the detection part 2 comprises a bottom plate 201, wherein a sample pad 202, a bonding pad 203, a nitrocellulose membrane 204 and a water absorption pad 205 are sequentially lapped and stuck on the bottom plate 201 from left to right, so that sample liquid dropped on the sample pad 202 sequentially flows to the water absorption pad 205 through the bonding pad 203 and the nitrocellulose membrane 204; the CTSZ antibody is sprayed on the bonding pad 203 and is used for being combined with an object to be detected in the sample liquid to form an object complex to be detected;
a display unit 3 including a detection line 301 and a quality control line 302; the detection line 301 and the quality control line 302 are arranged on the nitrocellulose membrane 204 along the flowing direction of the sample liquid, the detection line 301 is an anti-CTSZ antibody coated with a murine source, and a marker is formed when the complex of the to-be-detected object moves forwards under the chromatography effect; the quality control line 302 is goat anti-mouse IgG, and is used for forming a marker when an unbound analyte continues to advance;
the sample pad was soaked in a phosphate buffer of 0.02M pH7.1-7.5 containing 1% BSA and 0.1% casein by mass for one hour, and then dried at 35-40℃for 3 hours.
As shown in fig. 1 and 2, the detector comprises a protection part 1 sleeved on the outer side of a detection part 2; the protection part 1 comprises a box body 101; the middle part of the upper end of the box body 101 is provided with a round-corner rectangular observation window 102; the observation window 102 is positioned at the upper end of the nitrocellulose membrane 204, and the display part 3 is displayed through the observation window 102; the detection part 2 is protected by the whole box body 101, the influence of the outside on the detection part 2 is reduced, and the change of the display part 3 is observed through the observation window 102; the left side of the upper end of the box body 101 is provided with a conical placing window 103; a placing window 103 is positioned at the upper end of the sample pad 202 and is used for dripping sample liquid into the sample pad 202; by the conical design, the sample liquid can be prevented from overflowing when the sample liquid is dripped into the sample pad 202 through the placing window 103.
Wherein, as shown in fig. 3 and 4, the bonding pad 203 is formed of a polyester film; the surface of the bonding pad 203 is sprayed with a CTSZ antibody material layer, and lanthanide fluorescent microspheres are coupled on the CTSZ antibody material layer; the diameter of the fluorescent microsphere is 120mm;
taking 50 mu L of the coated fluorescent microsphere solution, adding EDC and MES buffer solution into the solution, wherein the final concentration of EDC is 0.05-0.2mg/ml, the final concentration of MES is 0.05-0.2mmol/L, oscillating for 15 minutes at room temperature, and centrifuging for fifteen minutes in a centrifugal tube flash refrigerated centrifuge at 18000 g/min; after removing the supernatant, adding a CTSZ antibody, wherein the mass ratio of the fluorescent microsphere to the CTSZ antibody is 1:300-1:100; after the reaction was carried out at room temperature for 1 hour and centrifuged at 18000g/min, the supernatant was removed, and then a BSA solution having a mass concentration of 1% was added to a final concentration of 0.05mol/L BSA.
The prepared fluorescent microsphere solution is sprayed on a polyester film by a metal spraying film drawing instrument in an amount of 2-5 mu L/cm, and the mixed microsphere solution is dried for one hour at 35-40 ℃ in the dark to obtain the bonding pad 203.
As shown in fig. 3, the interval between the quality control line 302 and the detection line 301 is set to be 5mm-8mm; the widths of the quality control line 302 and the detection line 301 are set to be 3mm-5mm;
the concentrations of the mouse anti-human IgG antibody and the sheep anti-mouse polyclonal antibody are respectively diluted to 1 mug/mu L by using 0.02mol/L phosphate buffer solution with pH of 7.1-7.5 and containing trehalose with the mass concentration of 1%, and the mouse anti-human IgG antibody and the sheep anti-mouse polyclonal antibody are respectively sprayed on the nitrocellulose membrane 204 by using a metal spraying and film scratching instrument in the amount of 1 mu L/cm, so as to obtain a detection line 301 and a quality control line 302 which are sequentially arranged.
Wherein, as shown in FIG. 5, the chromatography is directed from the conjugate pad 203 to the absorbent pad 205.
As shown in fig. 3, the absorbent pad 205 is absorbent paper.
And sequentially assembling the sample pad, the bonding pad, the nitrocellulose membrane and the absorbent paper prepared by the steps on a bottom plate to obtain the CTSZ antibody fluorescence immunochromatographic test paper.
Specific use and action of the embodiment: in the utility model, a fluorescent microsphere coated with Eu3+ is marked with CTSZ antibody, a metal spraying and film drawing instrument is used for spraying on a polyester film, after a sample liquid is added on a sample pad 202, the sample liquid swims to a water absorbing pad 205 for a section under the capillary action, and an object to be detected in the sample and an antigen Eu3+ -CTSZ antibody on the fluorescent microsphere form an object complex to be detected (Eu3+ -CTSZ antibody-CTSZ); with the chromatography, the complex moves forward, and reaches the detection line 301 for identifying the object to be detected (CTSZ) to form a secondary antibody-object to be detected-specific antibody sandwich complex (Eu3+ -mouse anti-human IgG-CTSZ antibody-CTSZ); aggregation of fluorescent microspheres occurs at detection line 301; when the unbound fluorescent microspheres continue to advance and reach the quality control line 302, the secondary antibodies (goat anti-mouse IgG) against the secondary antibodies bind with the secondary antibodies on the fluorescent microspheres, aggregation of the fluorescent microspheres occurs at the quality control line 302, and excessive secondary antibodies of the fluorescent microspheres continue to migrate to the water absorption pad 205; and a time-resolved fluorescence immunochromatography analyzer is adopted to read the detection line 301 and the quality control line 302 to generate corresponding fluorescence signals, and the concentration of the sample solution to be detected can be obtained by referring to a standard concentration curve.
The foregoing is merely exemplary embodiments of the present utility model and is not intended to limit the scope of the utility model, which is defined by the appended claims.
Claims (10)
1. A device for rapid detection of cathepsin Z, comprising:
the detection part (2) comprises a bottom plate (201), wherein the bottom plate (201) is sequentially lapped and stuck with a sample pad (202), a bonding pad (203), a nitrocellulose membrane (204) and a water absorption pad (205) from left to right, so that sample liquid dropped on the sample pad (202) sequentially passes through the bonding pad (203) and the nitrocellulose membrane (204) to swim towards the water absorption pad (205); the CTSZ antibody is sprayed on the bonding pad (203) and used for being combined with a part of the objects to be detected in the sample liquid to form the complex of the objects to be detected;
a display unit (3) comprising a detection line (301) and a quality control line (302); the detection line (301) and the quality control line (302) are arranged on the nitrocellulose membrane (204) along the flowing direction of the sample liquid, the detection line (301) is an anti-CTSZ antibody coated with a murine source and is used for forming a marker when the compound to be detected moves forwards under the chromatography effect; the quality control line (302) is goat anti-mouse IgG and is used for forming a marker when an unbound analyte continues to move forward.
2. The device for rapid detection of cathepsin Z according to claim 1, characterized in that it comprises a protection part (1) sleeved outside the detection part (2); the protection part (1) comprises a box body (101); the middle part of the upper end of the box body (101) is provided with a round-corner rectangular observation window (102); the observation window (102) is positioned at the upper end of the nitrocellulose membrane (204), and the display part (3) is displayed through the observation window (102).
3. The device for rapidly detecting cathepsin Z according to claim 2, wherein a conical placement window (103) is formed on the left side of the upper end of the box body (101); the placing window (103) is positioned at the upper end of the sample pad (202) and is used for dripping sample liquid into the sample pad (202).
4. The device for rapid detection of cathepsin Z according to claim 1, characterized in that the binding pad (203) is formed by a polyester film.
5. The device for rapid detection of cathepsin Z according to claim 4, wherein the surface of the binding pad (203) is sprayed with a layer of CTSZ antibody material, onto which lanthanide fluorescent microspheres are coupled.
6. The device for rapid detection of cathepsin Z of claim 5 wherein the fluorescent microsphere has a diameter of 120nm.
7. The device for rapidly detecting cathepsin Z according to claim 1, wherein the distance between the quality control line (302) and the detection line (301) is set to be 5mm-8mm.
8. The device for rapid detection of cathepsin Z according to claim 1, wherein the width of the quality control line (302) and the detection line (301) is set to 3mm-5mm.
9. A device for rapid detection of cathepsin Z according to claim 1, characterized in that the chromatography is directed from the conjugate pad (203) to the absorbent pad (205).
10. The device for rapid detection of cathepsin Z according to claim 1, wherein the absorbent pad (205) is absorbent paper.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202321926118.8U CN220323330U (en) | 2023-07-21 | 2023-07-21 | Device for rapidly detecting cathepsin Z |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202321926118.8U CN220323330U (en) | 2023-07-21 | 2023-07-21 | Device for rapidly detecting cathepsin Z |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN220323330U true CN220323330U (en) | 2024-01-09 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202321926118.8U Active CN220323330U (en) | 2023-07-21 | 2023-07-21 | Device for rapidly detecting cathepsin Z |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN220323330U (en) |
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2023
- 2023-07-21 CN CN202321926118.8U patent/CN220323330U/en active Active
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