CN107290547A - A kind of infertile quick detection kit and preparation method thereof - Google Patents
A kind of infertile quick detection kit and preparation method thereof Download PDFInfo
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- 238000005507 spraying Methods 0.000 claims description 8
- 238000002604 ultrasonography Methods 0.000 claims description 8
- 210000000481 breast Anatomy 0.000 claims description 5
- 102000011022 Chorionic Gonadotropin Human genes 0.000 claims description 4
- 108010062540 Chorionic Gonadotropin Proteins 0.000 claims description 4
- 230000000469 anti-sperm effect Effects 0.000 claims description 4
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- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 3
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Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- 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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/689—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to pregnancy or the gonads
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- 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
- G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
- G01N33/76—Human chorionic gonadotropin including luteinising hormone, follicle stimulating hormone, thyroid stimulating hormone or their receptors
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/36—Gynecology or obstetrics
- G01N2800/367—Infertility, e.g. sperm disorder, ovulatory dysfunction
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Medicinal Chemistry (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- Endocrinology (AREA)
- Biotechnology (AREA)
- Food Science & Technology (AREA)
- Reproductive Health (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Cell Biology (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Gynecology & Obstetrics (AREA)
- Pregnancy & Childbirth (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a kind of infertile quick detection kit and preparation method thereof, it is desirable to provide a kind of easy to operate, detects functional diversities, the reliable infertile quick detection kit of testing result and preparation method thereof；Technical points, including the box body with lid, described lid is provided with two wells；The sperm antibody detection card of the described box body microballoon arranged in parallel containing red latex, AEA ELISA detection card, the vitellary membrane antibody of yellow latex beads of the ovary antibody test card of pink colour latex beads, orange latex beads detect that the hCG antibodies of card, trophocyte's membrane antibody detection card of green latex beads and blue latex microballoon detect card；Belong to technical field of biological.
Description
Technical field
The invention discloses a kind of infertile detection kit, and in particular to a kind of infertile quick detection reagent
Box, the present invention also relates to the preparation method of the infertile quick detection kit.
Background technology
In recent years, due to environmental pollution exacerbation, lifestyle change, marriage and childbirth time retardation, sexually transmitted disease increase etc.
Influence, the infertile incidence of disease is in ascendant trend year by year.In industrialized country, infertile illness rate is about 8.5%-
20%.Correlation study shows that infertility occurs for about 8%-15% in the couple at child-bearing age of China, and its incidence of disease becomes in growth
Gesture.The World Health Organization predicts that infertility 21 century is by as being only second to tumour, the third-largest disease of cardiovascular disease.
It is infertile to be broadly divided into primary infertility and secondary infertility.Primary infertility is never to become pregnant；Secondary infertility is once to be pregnant
It is later and infertile.There are infertile, the intrauterine of ovulation failure, dysspermatism, oviduct abnormality, unknown cause the reason for causing infertile
Endometriosis and other such as immune infertilities.Immune infertility is the isoimmunization or autoimmunity by reproductive system antigen
Cause one of common disease and frequently-occurring disease, account for the 20-40% of a variety of causes infertility, and in recent years in the trend increased.Exempt from
Epidemic disease sexual factor cause it is infertile be the main of most of unknown sterility infertility, Secondary infertile and recurrent abortion
Reason, therefore, infertile detected caused to sexual factor is immunized also are necessary.
Infertile immunology diagnosis mainly includes AsAb (ASA, anti-spermatoz oa
Antibody), AOAb (AOA, anti-ovarian antibody), AEA (AEA, anti-
Endometrial antibody), anti-vitellary membrane antibody (AZP, anti-zonapel lucid antibody), anti-trophoderm
Cell membrane antibody (ATA, anti-trophoblastmcmbrane antibody), anti-hCG antibodies
Indexs such as (HCG, anti-humanchorionicgo nadotropin antibody).
Infertile detection method mainly has slide method, dot enzyme-linked immuno method and a colloidal gold method at present, but slide
Method, dot enzyme-linked immuno method need the steps such as cyclic washing, and operation is comparatively laborious, and the technical requirements to operating personnel are higher；Glue
Body gold method is simple to operate, but sensitivity is low, it is impossible to meet clinical needs.
Number of patent application 200810204669.7 provides a kind of protein chip detection method, can be to several antibody simultaneously
Detection, but it such as needs to wash, is incubated at step, it is cumbersome, it is necessary to which chemiluminescence instrument, is not suitable for basic unit and applies.Patent
Application number 201510444789.4 uses enzyme process, but cumbersome, and completing whole pattern detection needs 7 steps to complete, no
It is adapted to clinical practice.
The content of the invention
In view of the above-mentioned problems, it is an object of the invention to provide one kind is easy to operate, detecting functional diversities, testing result can
Infertile quick detection kit leaned on and preparation method thereof.
Therefore, first technical scheme that the present invention is provided is such：
A kind of infertile quick detection kit, including the box body with lid, described lid is provided with two wells
With a watch window；Sperm antibody detection card, the pink colour latex beads of the described box body microballoon arranged in parallel containing red latex
Ovary antibody test card, orange latex beads AEA ELISA detection card, yellow latex beads vitellary membrane antibody inspection
The human chorionic gonadotrophin for surveying card, trophocyte's membrane antibody detection card of green latex beads and blue latex microballoon resists
Card is surveyed in physical examination.
Further, a kind of above-mentioned infertile quick detection kit, the sperm of the microballoon containing red latex
Antibody test card, which includes successively being connected on the first bottom plate, the first described bottom plate, the first sample pad, goat anti-human igg antibody's mark
Red latex microballoon label pad, coated first coated film of sperm antigen and the first adsorptive pads, on the first described coated film
Provided with a sperm antigen detection zone T1 line and a first IgG quality control regions line C1 line, two lines are arranged in parallel.
Further, a kind of above-mentioned infertile quick detection kit, the ovary of the latex beads containing pink colour
Antibody test card, which includes successively being connected on the second bottom plate, the second described bottom plate, the second sample pad, goat anti-human igg antibody's mark
Pink colour latex beads label pad, the second coated film and the second adsorptive pads of ovary antigen coat, on the second described coated film
Provided with one bar of ovary antigen detection zone T2 line and one bar of IgG quality control region line C2 line of people the 2nd, two lines are arranged in parallel.
Further, above-mentioned a kind of infertile quick detection kit, the described uterus containing orange latex beads
Inner membrance antibody test card, which includes successively being connected on the 3rd bottom plate, the 3rd described bottom plate, the 3rd sample pad, goat anti-human igg antibody
Orange latex beads label pad, coated 3rd coated film of Endometrial Antigen and the 3rd adsorptive pads of mark, the described the 3rd
Coated film is provided with an Endometrial Antigen detection zone T3 line and third party's IgG quality control region C3 line, two lines parallel
Row.
Further, above-mentioned a kind of infertile quick detection kit, the latex beads containing yellow it is transparent
Band antibody test card, which includes successively being connected on the 4th bottom plate, the 4th described bottom plate, the 4th sample pad, goat anti-human igg antibody's mark
The yellow latex beads label pad of note, the 4th coated film and the 4th adsorptive pads of anti-oolemma antigen coat, the 4th described bag
Envelope is provided with an anti-oolemma antigen detection zone T4 line and a human IgG quality control region C4 line, and two lines are arranged in parallel.
Further, a kind of above-mentioned infertile quick detection kit, the nourishing containing green latex beads
Confluent monolayer cells membrane antibody detection card, which includes successively being connected on the 5th bottom plate, the 5th described bottom plate, the 5th sample pad, goat anti-human igg
Coated 5th coated film of the green latex beads label pad of antibody labeling, trophocyte's membranous antigen and the 5th adsorptive pads, institute
The 5th coated film stated is provided with one bar of trophocyte's membranous antigen detection zone T5 line and one article of the 5th human IgG quality control region C5 line,
Two lines are arranged in parallel.
Further, above-mentioned a kind of infertile quick detection kit, people's suede of the microballoon containing blue latex
Chorionic Gonadotropin antibody test card, which includes successively being connected on the 6th bottom plate, the 6th described bottom plate, the 6th sample pad, sheep
Anti-human IgG antibodies mark blue latex microballoon label pad, the 6th coated film of human chorionic gonadotrophin antigen coat and
6th adsorptive pads, the 6th described coated film is provided with one bar of human chorionic gonadotrophin T6 line and one article of Sixth Man IgG matter
Area's line is controlled, two lines are arranged in parallel.
Further, a kind of above-mentioned infertile quick detection kit, two described wells are located at lid
Lid above the same side, every three tactic detection cards has a well.
The latter solution that the present invention is provided is the preparation method of the kit：
A kind of preparation method of infertile quick detection kit, first prepares each detection card, then each by what is prepared
Individual detection card is positioned in box body, and each described detection blocking Preparation Method is that linking has corresponding sample successively on bottom plate
Pad, corresponding latex beads label pad, corresponding coated film and corresponding adsorptive pads：
The preparation method of the red latex microballoon label pad of described AsAb comprises the steps successively：
1. microballoon is pre-processed：Take particle diameter 200nm the μ L of red latex microballoon 10 be added to 0.1M pH5.5 1mL lemon
In lemon acid buffer, mix；
2. the activation of microballoon：The μ L of CMC (sanlose) 50 that concentration is 20mg/mL are taken, above-mentioned solution is added
In, mix；Reaction 10min is mixed on rotary incubator；
3. microballoon is quenched：Unnecessary CMC is quenched in the 2 mercapto ethanol for adding final concentration of 0.02M；
4. coupled antibody：Goat anti-human igg antibody 30ug is added into above-mentioned solution, reaction is mixed on rotary incubator
1h；
5. microballoon is closed：The OVA200 μ L of sealer 5% are added, reaction 30min is mixed on rotary incubator；
6. beads purification：Protein microsphere compound is separated from solution with desalting column；
7. preserve：The preservation liquid that volume is 200 μ L is added into desalting column, is moved in EP pipes, ultrasound is broken up 2 times；It is described
Preserve liquid for 0.1M pH8.0 glycine-NaOH buffer casein-sodium containing 1wt%, 10% trehalose, 2%
PEG20000,0.5%EDTA and 0.2mg/mL mouse IgG.
8. the solution of above-mentioned gained is drawn into film instrument with metal spraying to be sprayed onto on polyester cellulose film, discharge rate is 1~5 μ L/cm, is put into
37 DEG C of baking ovens, are dried 2~5 hours；
The preparation method of the pink colour latex beads label pad of described AOAb detection card resists with described anti-sperm
The preparation method difference that the red latex microballoon label pad of card is surveyed in physical examination is, grain is chosen in step 1. microballoon pretreatment
Footpath 200nm pink colour latex beads；
The preparation method and described anti-essence of the orange latex beads label pad of described AEA detection card
The preparation method difference of the red latex microballoon label pad of sub- antibody test card is, is selected in step 1. microballoon pretreatment
Take particle diameter 200nm orange latex beads；
The preparation method and described anti-sperm of the yellow latex beads label pad of described anti-vitellary membrane antibody detection card
The preparation method difference of the red latex microballoon label pad of antibody test card is, is chosen in step 1. microballoon pretreatment
Particle diameter 200nm yellow latex beads；
The preparation method of the green latex beads label pad of described anti-trophocyte's membrane antibody detection card with it is described
The preparation method difference of red latex microballoon label pad of AsAb detection card be that 1. microballoon is located in advance in step
Particle diameter 200nm green latex beads is chosen in reason；
The preparation method of the blue latex microballoon label pad of described anti-hCG antibodies detection card with
The preparation method difference of the red latex microballoon label pad of described AsAb detection card is, in step 1. microballoon
Particle diameter 200nm blue latex microballoon is chosen in pretreatment；
1. sperm antigen is coated with
Detection line：Rule on nitrocellulose membrane, the mark of C lines and T lines is carried out with marking pen in one end of nitrocellulose membrane
Note, 1.0mg/mL bags are diluted to the PBS containing 3% methanol, the 0.01M of 0.5% trehalose pH7.4 by sperm antigen
Quilt, line concentration is 1 μ L/cm, speed 100mm/s；
Nature controlling line：With containing 3% methanol, the 0.01M of 0.5% trehalose pH7.4 PBS is by human IgG polyclonal antibody
It is diluted to 0.8mg/mL to be coated with, line concentration is 1 μ L/cm, speed 100mm/s；
2. dry
37 DEG C of baking ovens are put into, are dried 6~8 hours；
The preparation method of the second described coated film and the preparation method difference of the first coated film are with containing 3%
Ovary antigen diluent is coated with by methanol, the 0.01M of 0.5% trehalose pH7.4 PBS to 1.0mg/mL；
The described preparation method of the 3rd coated film is to use 3% first with the preparation method difference of the first coated film
Endometrial Antigen is diluted to 1.0mg/mL and is coated with by alcohol, the 0.01M of 0.5% trehalose pH7.4 PBS；
The described preparation method of the 4th coated film is to use 3% first with the preparation method difference of the first coated film
Oolemma antigen diluent is coated with by alcohol, the 0.01M of 0.5% trehalose pH7.4 PBS to 1.0mg/mL；
The described preparation method of the 5th coated film is to use 3% first with the preparation method difference of the first coated film
Trophocyte's membranous antigen is diluted to 1.0mg/mL and is coated with by alcohol, the 0.01M of 0.5% trehalose pH7.4 PBS；
The described preparation method of the 6th coated film is to use 3% first with the preparation method difference of the first coated film
Alcohol, the 0.01M of 0.5% trehalose pH7.4 PBS is by human chorionic gonadotrophin antigen diluent to 1.0mg/mL bags
Quilt.
Further, a kind of preparation method of above-mentioned infertile quick detection kit, described sperm antigen comes
From in Seminal plasma；Described ovary antigen comes from monkey ovary；Described Endometrial Antigen comes from cattle uterus inner membrance；It is described
Oolemma antigen come from ox ovarian follicle；Described trophocyte's membranous antigen comes from human trophoblastic cells；It is described
Human chorionic gonadotrophin come from people's pregnant urine.
Compared with prior art, infertile quick detection kit provided by the present invention has the following advantages that：
1st, the technical scheme method that the present invention is provided detects six kinds of indexs using the color latex microballoon of six kinds of different colours,
It can judge which project detected according to the color of latex beads, it is to avoid mutually obscure between project；
2nd, lid of the technical scheme method that the present invention is provided above every three detection cards has a well, operates
When, it is only necessary to it can obtain six testing results in plus sample twice, 10min；
3rd, to efficiently solve infertile detection reagent in the market cumbersome for the technical scheme that provides of the present invention
Problem；The method that the present invention is provided only needs to 2 steps, simple to operate, and 10min can go out result, it is only necessary to plus sample is twice
6 projects are can detect, relatively low are required to operating personnel, are adapted to basic unit's application；
4th, the technical scheme that the present invention is provided is compared with colloidal gold method, and its sensitivity is higher than colloidal gold method, due to collaurum
Particle diameter it is smaller, general 40nm；Color latex microballoon, its particle diameter 200nm, sensitivity are used in the technical scheme that the present invention is provided
Height, better meets clinical practice.
Brief description of the drawings
Fig. 1 is the pregnant sterile six joint inspections quick detection kit structural representation that the present invention is provided；
Fig. 2 is the sperm antibody detection card A structural representations of red latex microballoon；
Fig. 3 is the ovary antibody test card B structure schematic diagram of pink colour latex beads；
Fig. 4 is the AEA ELISA detection card C-structure schematic diagram of orange latex beads；
Fig. 5 is the vitellary membrane antibody detection card D structural representations of yellow latex beads；
Fig. 6 is trophocyte's membrane antibody detection card E structural representations of green latex beads；
Fig. 7 is the hCG antibodies detection card F structural representations of blue latex microballoon.
Embodiment
It is further to the claim of the present invention to be limited with reference to embodiment, but do not constitute to this
Any limitation of invention.
A kind of infertile quick detection kit that the present invention is provided refers to Fig. 1 to Fig. 7, including the box body with lid 11
1, described lid 11 is provided with two wells 12 and an observation window 13, and two described wells 12 are located at the same of lid 11
Side, the width sums of the distance between two wells 12 with two detection cards, wherein the lids above every three detections card
11 have a well 12, during operation, it is only necessary to plus sample twice, and six testing results can be obtained in 10min；Described box body
Sperm antibody detection card A, ovary antibody test card B, the orange breast of pink colour latex beads of 1 microballoon arranged in parallel containing red latex
The AEA ELISA detection card C of glue microballoon, the vitellary membrane antibody detection card D of yellow latex beads, the taste of green latex beads
Support the hCG antibodies detection card F of confluent monolayer cells membrane antibody detection card E and blue latex microballoon.
More specifically, the sperm antibody detection card A of the microballoon containing red latex includes the first bottom plate 1a, described
The first bottom plate 1a on successively linking have the first sample pad 2a, goat anti-human igg antibody mark red latex microballoon label pad 3a,
The coated first coated film 4a of sperm antigen and the first adsorptive pads 5a, the first described coated film 4a is provided with a sperm antigen
Detection zone T1 lines and a first IgG quality control regions C1 line, two lines are arranged in parallel.
More specifically, the ovary antibody test card B of the latex beads containing pink colour includes the second bottom plate 1b, described
The second bottom plate 1b on successively linking have the second sample pad 2b, goat anti-human igg antibody mark pink colour latex beads label pad 3b,
Second coated film 4b of ovary antigen coat and the second adsorptive pads 5b, the second described coated film 4b is provided with an ovary antigen
Detection zone T1 lines and one bar of IgG quality control region C2 line of people the 2nd, two lines are arranged in parallel.
More specifically, the described detection card of the AEA ELISA containing orange latex beads C includes the 3rd bottom plate 1c,
Linking has the orange latex beads mark of the 3rd sample pad 2c, goat anti-human igg antibody's mark successively on the 3rd described bottom plate 1c
Pad 3c, Endometrial Antigen coated 3rd coated film 4c and the 3rd adsorptive pads 5c, the 3rd described coated film 4c and be provided with one
Bar Endometrial Antigen detection zone T3 lines and third party's IgG quality control region C3 line, two lines are arranged in parallel.
More specifically, the vitellary membrane antibody detection card D of the latex beads containing yellow includes the 4th bottom plate 1d, institute
Linking has the 4th sample pad 2d, the yellow latex beads label pad of goat anti-human igg antibody's mark successively on the 4th bottom plate 1d stated
3d, anti-oolemma antigen coat the 4th coated film 4d and the 4th adsorptive pads 5d, the 4th described coated film 4d are provided with one article
Sperm antigen detection zone T5 lines and a human IgG quality control region C5 line, two lines are arranged in parallel.
More specifically, trophocyte's membrane antibody detection card E containing green latex beads includes the 5th bottom
Linking has the 5th sample pad 2e, the green latex beads of goat anti-human igg antibody's mark successively on plate 1e, the 5th described bottom plate 1e
Label pad 3e, trophocyte's membranous antigen coated 5th coated film 4e and the 5th adsorptive pads 5e, the 5th described coated film 4e
One bar of sperm antigen detection zone T line and one article of the 5th human IgG quality control region C line are provided with, two lines are arranged in parallel.
More specifically, trophocyte's membrane antibody detection card F of the microballoon containing blue latex includes the 6th bottom
Linking has the 6th sample pad 2f, the blue latex microballoon of goat anti-human igg antibody's mark successively on plate 1f, the 6th described bottom plate 1f
Label pad 3f, human chorionic gonadotrophin antigen coat the 6th coated film 4f and the 6th adsorptive pads 5f, the 6th described bag
Envelope 4f is provided with a human chorionic gonadotrophin T6 line and a Sixth Man IgG quality control region C line, two lines parallel
Row.
A kind of preparation method of above-mentioned infertile quick detection kit, first prepares each detection card, then will prepare
Each good detection card is positioned in box body 1.
Described sperm antibody detection card A comprises the steps successively：It is connected the first sample successively on the first bottom plate 1a
Pad 2a, red latex microballoon label pad 3a, the coated first coated film 4a and first of sperm antigen of goat anti-human igg antibody's mark
Adsorptive pads 5a, wherein：
1) the first sample pad 2a preparation method is：Sample pad is cut into 20mm × 300mm size, sample pad is immersed in
In buffer solution, take out after 1 hour, in drying at room temperature 16-18 hours.
First sample pad 1b buffer formulation is as follows：2%BSA, 1%PVP, 0.5%Tween are dissolved in 0.01 PBS
(pH7.4) in buffer solution.
The red latex microballoon label pad 2a of described AsAb detection card preparation method includes following steps successively
Suddenly：
1. microballoon is pre-processed：Take particle diameter 200nm the μ L of red latex microballoon 10 be added to 0.1M pH5.5 1mL lemon
In lemon acid buffer, mix；
2. the activation of microballoon：The CMC50 μ L that concentration is 20mg/mL are taken, are added in above-mentioned solution, are mixed；In rotating and culturing
Reaction 10min is mixed on device；
3. microballoon is quenched：Add final concentration of 0.02M 2 mercapto ethanol be quenched it is unnecessary
CMC；
4. coupled antibody：Goat anti-human igg antibody 30ug is added into above-mentioned solution, reaction is mixed on rotary incubator
1h；
5. microballoon is closed：The OVA200 μ L of sealer 5% are added, reaction 30min is mixed on rotary incubator；
6. beads purification：Protein microsphere compound is separated from solution with desalting column；
7. preserve：The preservation liquid that volume is 200 μ L is added into desalting column, is moved in EP pipes, ultrasound is broken up 2 times；It is described
Preserve liquid for 0.1M pH8.0 glycine-NaOH buffer casein-sodium containing 1wt%, 10% trehalose, 2%
PEG20000,0.5%EDTA and 0.2mg/mL mouse IgG.
8. the solution of above-mentioned gained is drawn into film instrument with metal spraying to be sprayed onto on polyester cellulose film, discharge rate is 1~5 μ L/cm, is put into
37 DEG C of baking ovens, are dried 2~5 hours；
3) the coated first coated film 4a preparation methods of sperm antigen are：
1. sperm antigen is coated with
Detection line (T1 lines)：Rule on nitrocellulose membrane, the mark of C1 lines and T1 lines is carried out with marking pen in one end of film
The distance of note, C lines and T lines is 0.5cm, and the distance of T lines and nitrocellulose filter lower edge is 1cm, uses 3% methanol, 0.5% marine alga
Sperm antigen is diluted to 1.0mg/mL and is coated with by the 0.01M of sugar pH7.4 PBS；Described sperm antigen comes from people
Seminal fluid, line concentration is 1 μ L/cm, speed 100mm/s.
Nature controlling line (C1 lines)：Use 3% methanol, the 0.01M of 0.5% trehalose pH7.4 PBS is by human IgG Anti-TNF-α
Body is diluted to 0.8mg/mL and is coated with, and line concentration is 1 μ L/cm, speed 100mm/s.
The ovary antibody test card B of described pink colour latex beads comprises the steps successively：On the second bottom plate 1b according to
The second sample pad 2b of secondary linking, the pink colour latex beads label pad 3b of goat anti-human igg antibody's mark, the second of ovary antigen coat
Coated film 4b and the second adsorptive pads 5b, wherein：
1) the second sample pad 2b preparation method is：Sample pad is cut into 20mm × 300mm size, sample pad is immersed in
In buffer solution, take out after 1 hour, in drying at room temperature 16-18 hours.
Second sample pad 1b buffer formulation is as follows：Buffer formulation is as follows：2%BSA, 1%PVP, 0.5%Tween
It is dissolved in 0.01 PBS (pH7.4) buffer solution.
The pink colour latex beads label pad 2b of described AOAb detection card preparation method includes following steps successively
Suddenly：
1. microballoon is pre-processed：Take particle diameter 200nm the μ L of pink colour latex beads 10 be added to 0.1M pH5.5 1mL lemon
In lemon acid buffer, mix；
2. the activation of microballoon：The CMC50 μ L that concentration is 20mg/mL are taken, are added in above-mentioned solution, are mixed；In rotating and culturing
Reaction 10min is mixed on device；
3. microballoon is quenched：Unnecessary CMC is quenched in the 2 mercapto ethanol for adding final concentration of 0.02M；
4. coupled antibody：Goat anti-human igg antibody 30ug is added into above-mentioned solution, reaction is mixed on rotary incubator
1h；
5. microballoon is closed：The OVA200 μ L of sealer 5% are added, reaction 30min is mixed on rotary incubator；
6. beads purification：Protein microsphere compound is separated from solution with desalting column；
7. preserve：The preservation liquid that volume is 200 μ L is added into desalting column, is moved in EP pipes, ultrasound is broken up 2 times；It is described
Preserve liquid for 0.1M pH8.0 glycine-NaOH buffer casein-sodium containing 1wt%, 10% trehalose, 2%
PEG20000,0.5%EDTA and 0.2mg/mL mouse IgG.
8. the solution of above-mentioned gained is drawn into film instrument with metal spraying to be sprayed onto on polyester cellulose film, discharge rate is 1~5 μ L/cm, is put into
37 DEG C of baking ovens, are dried 2~5 hours；
3) the second coated film 4b preparation methods of ovary antigen coat are：
1. ovary antigen coat
Detection line (T2 lines)：Rule on nitrocellulose membrane, the mark of C lines and T lines carried out with marking pen in one end of film,
The distance of C2 lines and T2 lines is 0.5cm, and the distance of T lines and nitrocellulose filter lower edge is 1cm, uses 3% methanol, 0.5% marine alga
Ovary antigen diluent is coated with by the 0.01M of sugar pH7.4 PBS to 1.0mg/mL；Described ovary antigen comes from monkey
Ovary, line concentration is 1 μ L/cm, speed 100mm/s.
Nature controlling line (C2 lines)：Use 3% methanol, the 0.01M of 0.5% trehalose pH7.4 PBS is by human IgG Anti-TNF-α
Body is diluted to 0.8mg/mL and is coated with, and line concentration is 1 μ L/cm, speed 100mm/s.
The AEA ELISA detection card C of described orange latex beads comprises the steps successively：In the 3rd bottom plate 1c
On be connected successively the 3rd sample pad 2c, goat anti-human igg antibody mark orange latex beads label pad 3c, Endometrial Antigen bag
3rd coated film 4c of quilt and the 3rd adsorptive pads 5c, wherein：
1) the 3rd sample pad 2c preparation method is：Sample pad is cut into 20mm × 300mm size, sample pad is immersed in
In buffer solution, take out after 1 hour, in drying at room temperature 16-18 hours.
3rd sample pad 1c buffer formulation is as follows：2%BSA, 1%PVP, 0.5%Tween are dissolved in 0.01 PBS
(pH7.4) in buffer solution.
Under the orange latex beads label pad 2c of described AEA detection card preparation method includes successively
State step：
1. microballoon is pre-processed：Take particle diameter 200nm the μ L of orange latex beads 10 be added to 0.1M pH5.5 1mL lemon
In lemon acid buffer, mix；
2. the activation of microballoon：The CMC50 μ L that concentration is 20mg/mL are taken, are added in above-mentioned solution, are mixed；In rotating and culturing
Reaction 10min is mixed on device；
3. microballoon is quenched：Unnecessary CMC is quenched in the 2 mercapto ethanol for adding final concentration of 0.02M；
4. coupled antibody：Goat anti-human igg antibody 30ug is added into above-mentioned solution, reaction is mixed on rotary incubator
1h；
5. microballoon is closed：The OVA200 μ L of sealer 5% are added, reaction 30min is mixed on rotary incubator；
6. beads purification：Protein microsphere compound is separated from solution with desalting column；
7. preserve：The preservation liquid that volume is 200 μ L is added into desalting column, is moved in EP pipes, ultrasound is broken up 2 times；It is described
Preserve liquid for 0.1M pH8.0 glycine-NaOH buffer casein-sodium containing 1wt%, 10% trehalose, 2%
PEG20000,0.5%EDTA and 0.2mg/mL mouse IgG.
8. the solution of above-mentioned gained is drawn into film instrument with metal spraying to be sprayed onto on polyester cellulose film, discharge rate is 1~5 μ L/cm, is put into
37 DEG C of baking ovens, are dried 2~5 hours；
3) the coated 3rd coated film 4c preparation methods of Endometrial Antigen are：
1. Endometrial Antigen is coated with
Detection line (T3 lines)：Rule on nitrocellulose membrane, the mark of C3 lines and T3 lines is carried out with marking pen in one end of film
The distance of note, C3 lines and T3 lines is 0.5cm, and the distance of T lines and nitrocellulose filter lower edge is 1cm, uses 3% methanol, 0.5% sea
Endometrial Antigen is diluted to 1.0mg/mL and is coated with by the 0.01M of algae sugar pH7.4 PBS；Described endometrium resists
Original comes from cattle uterus inner membrance, and line concentration is 1 μ L/cm, speed 100mm/s.
Nature controlling line (C3 lines)：Use 3% methanol, the 0.01M of 0.5% trehalose pH7.4 PBS is by human IgG Anti-TNF-α
Body is diluted to 0.8mg/mL and is coated with, and line concentration is 1 μ L/cm, speed 100mm/s.
The vitellary membrane antibody detection of described yellow latex beads blocks D, comprised the steps successively：On the 4th bottom plate 1d
The 4th sample pad 2d of linking, the yellow latex beads label pad 3d of goat anti-human igg antibody's mark, oolemma antigen coat successively
4th coated film 4d and the 4th adsorptive pads 5d, wherein：
1) the 4th sample pad 2d preparation method is：Sample pad is cut into 20mm × 300mm size, sample pad is immersed in
In buffer solution, take out after 1 hour, in drying at room temperature 16-18 hours.
4th sample pad 1d buffer formulation is as follows：2%BSA, 1%PVP, 0.5%Tween are dissolved in 0.01 PBS
(pH7.4) in buffer solution.
The yellow latex beads label pad 2d of described anti-vitellary membrane antibody detection card preparation method includes following successively
Step：
1. microballoon is pre-processed：Take particle diameter 200nm the μ L of yellow latex beads 10 be added to 0.1M pH5.5 1mL lemon
In lemon acid buffer, mix；
2. the activation of microballoon：The CMC50 μ L that concentration is 20mg/mL are taken, are added in above-mentioned solution, are mixed；In rotating and culturing
Reaction 10min is mixed on device；
3. microballoon is quenched：Unnecessary CMC is quenched in the 2 mercapto ethanol for adding final concentration of 0.02M；
4. coupled antibody：Goat anti-human igg antibody 30ug is added into above-mentioned solution, reaction is mixed on rotary incubator
1h；
5. microballoon is closed：The OVA200 μ L of sealer 5% are added, reaction 30min is mixed on rotary incubator；
6. beads purification：Protein microsphere compound is separated from solution with desalting column；
7. preserve：The preservation liquid that volume is 200 μ L is added into desalting column, is moved in EP pipes, ultrasound is broken up 2 times；It is described
Preserve liquid for 0.1M pH8.0 glycine-NaOH buffer casein-sodium containing 1wt%, 10% trehalose, 2%
PEG20000,0.5%EDTA and 0.2mg/mL mouse IgG.
8. the solution of above-mentioned gained is drawn into film instrument with metal spraying to be sprayed onto on polyester cellulose film, discharge rate is 1~5 μ L/cm, is put into
37 DEG C of baking ovens, are dried 2~5 hours；
3) the 4th coated film 4d preparation methods of oolemma antigen coat are：
1. oolemma antigen coat
Detection line (T4 lines)：Rule on nitrocellulose membrane, the mark of C4 lines and T4 lines is carried out with marking pen in one end of film
The distance of note, C4 lines and T4 lines is 0.5dm, and the distance of T lines and nitrocellulose filter lower edge is 1cm, uses 3% methanol, 0.5% sea
Oolemma antigen diluent is coated with by the 0.01M of algae sugar pH7.4 PBS to 1.0mg/mL；Described oolemma antigen comes
Come from ox ovarian follicle from oolemma antigen, line concentration is 1 μ L/cm, speed 100mm/s.
Nature controlling line (D lines)：Use 3% methanol, the 0.01M of 0.5% trehalose pH7.4 PBS is by human IgG polyclonal antibody
It is diluted to 0.8mg/mL to be coated with, line concentration is 1 μ L/cm, speed 100mm/s.
Trophocyte's membrane antibody detection of described green latex beads blocks E, comprised the steps successively：At the 5th bottom
It is connected the 5th sample pad 2e, green latex beads label pad 3e, the trophocyte of goat anti-human igg antibody's mark on plate 1e successively
The coated 5th coated film 4e and the 5th adsorptive pads 5e of membranous antigen, wherein：
1) the 5th sample pad 2e preparation method is：Sample pad is cut into 20mm × 300mm size, sample pad is immersed in
In buffer solution, take out after 1 hour, in drying at room temperature 16-18 hours.
5th sample pad 1e buffer formulation is as follows：2%BSA, 1%PVP, 0.5%Tween are dissolved in 0.01 PBS
(pH7.4) in buffer solution.
The green latex beads label pad 2e of described anti-trophocyte's membrane antibody detection card preparation method is wrapped successively
Include following step：
1. microballoon is pre-processed：Take particle diameter 200nm the μ L of green latex beads 10 be added to 0.1M pH5.5 1mL lemon
In lemon acid buffer, mix；
2. the activation of microballoon：The CMC50 μ L that concentration is 20mg/mL are taken, are added in above-mentioned solution, are mixed；In rotating and culturing
Reaction 10min is mixed on device；
3. microballoon is quenched：Unnecessary CMC is quenched in the 2 mercapto ethanol for adding final concentration of 0.02M；
4. coupled antibody：Goat anti-human igg antibody 30ug is added into above-mentioned solution, reaction is mixed on rotary incubator
1h；
5. microballoon is closed：The OVA200 μ L of sealer 5% are added, reaction 30min is mixed on rotary incubator；
6. beads purification：Protein microsphere compound is separated from solution with desalting column；
7. preserve：The preservation liquid that volume is 200 μ L is added into desalting column, is moved in EP pipes, ultrasound is broken up 2 times；It is described
Preserve liquid for 0.1M pH8.0 glycine-NaOH buffer casein-sodium containing 1wt%, 10% trehalose, 2%
PEG20000,0.5%EDTA and 0.2mg/mL mouse IgG.
8. the solution of above-mentioned gained is drawn into film instrument with metal spraying to be sprayed onto on polyester cellulose film, discharge rate is 1~5 μ L/cm, is put into
37 DEG C of baking ovens, are dried 2~5 hours；
3) the coated 5th coated film 4e preparation methods of trophocyte's membranous antigen are：
1. trophocyte's membranous antigen is coated with
Detection line (T5 lines)：Rule on nitrocellulose membrane, the mark of C5 lines and T5 lines is carried out with marking pen in one end of film
The distance of note, C5 lines and T5 lines is 0.5cm, and the distance of T lines and nitrocellulose filter lower edge is 1cm, uses 3% methanol, 0.5% sea
Trophocyte's membranous antigen is diluted to 1.0mg/mL and is coated with by the 0.01M of algae sugar pH7.4 PBS；Described trophoderm
The anti-trophocyte's membranous antigen of cell membrane comes from human trophoblastic cells, and line concentration is 1 μ L/cm, speed 100mm/
s。
Nature controlling line (C5 lines)：Use 3% methanol, the 0.01M of 0.5% trehalose pH7.4 PBS is by human IgG Anti-TNF-α
Body is diluted to 0.8mg/mL and is coated with, and line concentration is 1 μ L/cm, speed 100mm/s.
The hCG antibodies detection of described blue latex microballoon blocks F, comprised the steps successively：
It is connected the 6th sample pad 2f, blue latex microballoon label pad 3f, the people's suede of goat anti-human igg antibody's mark on 6th bottom plate 1f successively
6th coated film 4f of Chorionic Gonadotropin antigen coat and the 6th adsorptive pads 5f, wherein：
1) the 6th sample pad 2f preparation method is：Sample pad is cut into 20mm × 300mm size, sample pad is immersed in
In buffer solution, take out after 1 hour, in drying at room temperature 16-18 hours.
6th sample pad 1f buffer formulation is as follows：2%BSA, 1%PVP, 0.5%Tween are dissolved in 0.01 PBS
(pH7.4) in buffer solution.
The blue latex microballoon label pad 2f of described anti-hCG antibodies detection card preparation method
Comprise the steps successively：
1. microballoon is pre-processed：Take particle diameter 200nm the μ L of blue latex microballoon 10 be added to 0.1M pH5.5 1mL lemon
In lemon acid buffer, mix；
2. the activation of microballoon：The CMC50 μ L that concentration is 20mg/mL are taken, are added in above-mentioned solution, are mixed；In rotating and culturing
Reaction 10min is mixed on device；
3. microballoon is quenched：Unnecessary CMC is quenched in the 2 mercapto ethanol for adding final concentration of 0.02M；
4. coupled antibody：Goat anti-human igg antibody 30ug is added into above-mentioned solution, reaction is mixed on rotary incubator
1h；
5. microballoon is closed：The OVA200 μ L of sealer 5% are added, reaction 30min is mixed on rotary incubator；
6. beads purification：Protein microsphere compound is separated from solution with desalting column；
7. preserve：The preservation liquid that volume is 200 μ L is added into desalting column, is moved in EP pipes, ultrasound is broken up 2 times；It is described
Preserve liquid for 0.1M pH8.0 glycine-NaOH buffer casein-sodium containing 1wt%, 10% trehalose, 2%
PEG20000,0.5%EDTA and 0.2mg/mL mouse IgG.
8. the solution of above-mentioned gained is drawn into film instrument with metal spraying to be sprayed onto on polyester cellulose film, discharge rate is 1~5 μ L/cm, is put into
37 DEG C of baking ovens, are dried 2~5 hours；
3) the 6th coated film 4f preparation methods of human chorionic gonadotrophin antigen coat are：
1. human chorionic gonadotrophin antigen coat
Detection line (T6 lines)：Rule on nitrocellulose membrane, the mark of C6 lines and T6 lines is carried out with marking pen in one end of film
The distance of note, C6 lines and T6 lines is 0.5cm, and the distance of T lines and nitrocellulose filter lower edge is 1cm, uses 3% methanol, 0.5% sea
Human chorionic gonadotrophin antigen diluent is coated with by the 0.01M of algae sugar pH7.4 PBS to 1.0mg/mL；Described
Human chorionic gonadotrophin comes from people's pregnant urine, and line concentration is 1 μ L/cm, speed 100mm/s.
Nature controlling line (C6 lines)：Use 3% methanol, the 0.01M of 0.5% trehalose pH7.4 PBS is by human IgG Anti-TNF-α
Body is diluted to 0.8mg/mL and is coated with, and line concentration is
| Project name | Detection line | Negative findings | Positive findings |
1 μ L/cm, speed 100mm/s.
For the infertile quick detection kit preferably provided using the present invention, offer of the present invention is given below
Sample testing method：
1. take in 100 μ L plasma/serums, the PBS dilutions for being added to 900 μ L；
2. each well respectively adds the 200 above-mentioned solution of μ L；
Result is read during 3.10min, as a result such as following table.
Above-described embodiment is preferably embodiment, but embodiments of the present invention are not by above-described embodiment of the invention
Limitation, other any Spirit Essences without departing from the present invention and the change made under principle, modification, replacement, combine, simplification,
Equivalent substitute mode is should be, is included within protection scope of the present invention.
Claims (10)
1. a kind of infertile quick detection kit, including the box body (1) with lid (11), it is characterised in that described box
Cover (11) and be provided with two wells (12) and a watch window (13)；Described box body (1) is arranged in parallel micro- containing red latex
Sperm antibody detection card (A), ovary antibody test card (B), the endometrium of orange latex beads of pink colour latex beads of ball
Antibody test card (C), the vitellary membrane antibody detection card (D) of yellow latex beads, trophocyte's film of green latex beads resist
The hCG antibodies detection card (F) of card (E) and blue latex microballoon is surveyed in physical examination.
2. a kind of infertile quick detection kit according to claim 1, it is characterised in that described breast containing red
The sperm antibody detection card (A) of glue microballoon, which includes successively being connected on the first bottom plate (1a), described the first bottom plate (1a), first
Sample pad (2a), the red latex microballoon label pad (3a) of goat anti-human igg antibody's mark, coated first coated film of sperm antigen
(4a) and the first adsorptive pads (5a), described the first coated film (4a) is provided with one bar of sperm antigen detection zone T1 line and one article the
One human IgG quality control region C1 lines, two lines are arranged in parallel.
3. a kind of infertile quick detection kit according to claim 1, it is characterised in that described is newborn containing pink colour
The ovary antibody test card (B) of glue microballoon, which includes successively being connected on the second bottom plate (1b), described the second bottom plate (1b), second
Sample pad (2b), the pink colour latex beads label pad (3b) of goat anti-human igg antibody's mark, the second coated film of ovary antigen coat
(4b) and the second adsorptive pads (5b), described the second coated film (4b) is provided with an ovary antigen detection zone T1 line and a people
2nd IgG quality control region C2 lines, two lines are arranged in parallel.
4. a kind of infertile quick detection kit according to claim 1, it is characterised in that described contains orange breast
The AEA ELISA detection card (C) of glue microballoon, which includes successively being connected on the 3rd bottom plate (1c), the 3rd described bottom plate (1c), to be had
3rd sample pad (2c), the orange latex beads label pad (3c) of goat anti-human igg antibody's mark, Endometrial Antigen coated the
Three coated films (4c) and the 3rd adsorptive pads (5c), the 3rd described coated film (4c) is provided with one article of Endometrial Antigen detection zone
T3 lines and third party's IgG quality control region C3 line, two lines are arranged in parallel.
5. a kind of infertile quick detection kit according to claim 1, it is characterised in that described is newborn containing yellow
The vitellary membrane antibody detection card (D) of glue microballoon includes the 4th bottom plate (1d), and linking has the successively on the 4th described bottom plate (1d)
Four sample pads (2d), the yellow latex beads label pad (3d) of goat anti-human igg antibody's mark, the 4th of anti-oolemma antigen coat the
Coated film (4d) and the 4th adsorptive pads (5d), the 4th described coated film (4d) are provided with one bar of oolemma detection zone T4 line and one
Bar human IgG quality control region C5 lines, two lines are arranged in parallel.
6. a kind of infertile quick detection kit according to claim 1, it is characterised in that described breast containing green
Trophocyte's membrane antibody detection card (E) of glue microballoon includes holding in the mouth successively on the 5th bottom plate (1e), the 5th described bottom plate (1e)
It is connected to the 5th sample pad (2e), the green latex beads label pad (3e) of goat anti-human igg antibody's mark, trophocyte's membranous antigen
Coated 5th coated film (4e) and the 5th adsorptive pads (5e), the 5th described coated film (4e) is provided with one article of trophocyte
Membranous antigen detection zone T5 lines and one article of the 5th human IgG quality control region C line, two lines are arranged in parallel.
7. a kind of infertile quick detection kit according to claim 1, it is characterised in that described breast containing blueness
Trophocyte's membrane antibody detection card (F) of glue microballoon includes holding in the mouth successively on the 6th bottom plate (1f), the 6th described bottom plate (1f)
The 6th sample pad (2f), the blue latex microballoon label pad (3f) of goat anti-human igg antibody's mark, human chorionic gonadotropin's gland is connected to swash
6th coated film (4f) of plain antigen coat and the 6th adsorptive pads (5f), the 6th described coated film (4f) is provided with one article of people's suede
Chorionic Gonadotropin antigen detection zone T6 lines and a Sixth Man IgG quality control region C line, two lines are arranged in parallel.
8. a kind of infertile quick detection kit according to claim 1, it is characterised in that two described sample-addings
The lid (11) that hole (12) is located above the same side of lid (11), every three tactic detection cards has a well
(12)。
9. a kind of preparation method of infertile quick detection kit described in claim 1, first prepares each detection card, then
Each detection card prepared is positioned in box body (1), each described detection blocking Preparation Method is held in the mouth successively on bottom plate
It is connected to corresponding sample pad, corresponding latex beads label pad, corresponding coated film and corresponding adsorptive pads, it is characterised in that
The preparation method of the red latex microballoon label pad of described AsAb detection card comprises the steps successively：
1. microballoon is pre-processed：Take particle diameter 200nm the μ L of red latex microballoon 10 be added to 0.1M pH5.5 1mL citric acid
In buffer solution, mix；
2. the activation of microballoon：The μ L of CMC 50 that concentration is 20mg/mL are taken, are added in above-mentioned solution, are mixed；On rotary incubator
Mix reaction 10min；
3. microballoon is quenched：Unnecessary CMC is quenched in the 2 mercapto ethanol for adding final concentration of 0.02M；
4. coupled antibody：Goat anti-human igg antibody 30ug is added into above-mentioned solution, reaction 1h is mixed on rotary incubator；
5. microballoon is closed：The OVA200 μ L of sealer 5% are added, reaction 30min is mixed on rotary incubator；
6. beads purification：Protein microsphere compound is separated from solution with desalting column；
7. preserve：The preservation liquid that volume is 200 μ L is added into desalting column, is moved in EP pipes, ultrasound is broken up 2 times；Described guarantor
Liquid storage for 0.1M pH8.0 glycine-NaOH buffer casein-sodium containing 1wt%, 10% trehalose, 2%PEG20000,
0.5%EDTA and 0.2mg/mL mouse IgG.
8. the solution of above-mentioned gained is drawn into film instrument with metal spraying to be sprayed onto on polyester cellulose film, discharge rate is 1~5 μ L/cm, is put into 37 DEG C
Baking oven, is dried 2~5 hours；
The preparation method of the pink colour latex beads label pad (3b) of described AOAb detection card resists with described anti-sperm
The preparation method difference that the red latex microballoon label pad of card is surveyed in physical examination is, particle diameter is chosen in the pretreatment of 1. microballoon
200nm pink colour latex beads；
The preparation method and described anti-essence of the orange latex beads label pad (3c) of described AEA detection card
The preparation method difference of the red latex microballoon label pad of sub- antibody test card is, grain is chosen in the pretreatment of 1. microballoon
Footpath 200nm orange latex beads；
The preparation method and described anti-sperm of the yellow latex beads label pad (3d) of described anti-vitellary membrane antibody detection card
The preparation method difference of the red latex microballoon label pad of antibody test card is, particle diameter is chosen in the pretreatment of 1. microballoon
200nm yellow latex beads；
The preparation method of the green latex beads label pad (3e) of described anti-trophocyte's membrane antibody detection card with it is described
The preparation method difference of red latex microballoon label pad of AsAb detection card be, in the pretreatment of 1. microballoon
Choose particle diameter 200nm green latex beads；
The preparation method of the blue latex microballoon label pad (3f) of described anti-hCG antibodies detection card with
The preparation method difference of the red latex microballoon label pad of described AsAb detection card is, locates in advance in 1. microballoon
Particle diameter 200nm blue latex microballoon is chosen in reason；
The preparation method of the first described coated film comprises the steps：
1. sperm antigen is coated with
Detection line：Rule on nitrocellulose membrane, carry out the mark of C lines and T lines with marking pen in one end of nitrocellulose membrane, used
Sperm antigen is diluted to 1.0mg/mL and is coated with by 3% methanol, the 0.01M of 0.5% trehalose pH7.4 PBS, is rule dense
Spend for 1 μ L/cm, speed 100mm/s；
Nature controlling line：3% methanol is used, human IgG polyclonal antibody is diluted to by the 0.01M of 0.5% trehalose pH7.4 PBS
0.8mg/mL is coated with, and line concentration is 1 μ L/cm, speed 100mm/s；
2. dry
37 DEG C of baking ovens are put into, are dried 6~8 hours；
The preparation method of the second described coated film and the preparation method difference of the first coated film be to use 3% methanol,
Ovary antigen diluent is coated with by the 0.01M of 0.5% trehalose pH7.4 PBS to 1.0mg/mL；
The described preparation method of the 3rd coated film is to use 3% methanol with the preparation method difference of the first coated film,
Endometrial Antigen is diluted to 1.0mg/mL and is coated with by the 0.01M of 0.5% trehalose pH7.4 PBS；
The described preparation method of the 4th coated film is to use 3% methanol with the preparation method difference of the first coated film,
Oolemma antigen diluent is coated with by the 0.01M of 0.5% trehalose pH7.4 PBS to 1.0mg/mL；
The described preparation method of the 5th coated film is to use 3% methanol with the preparation method difference of the first coated film,
Trophocyte's membranous antigen is diluted to 1.0mg/mL and is coated with by the 0.01M of 0.5% trehalose pH7.4 PBS；
The described preparation method of the 6th coated film is to use 3% methanol with the preparation method difference of the first coated film,
Human chorionic gonadotrophin antigen diluent is coated with by the 0.01M of 0.5% trehalose pH7.4 PBS to 1.0mg/mL.
10. a kind of preparation method of infertile quick detection kit according to claim 9, it is characterised in that institute
The sperm antigen stated comes from Seminal plasma；Described ovary antigen comes from monkey ovary；Described Endometrial Antigen comes from
Cattle uterus inner membrance；Described oolemma antigen comes from ox ovarian follicle；Described trophocyte's membranous antigen comes from human chorionic
Trophocyte；Described human chorionic gonadotrophin comes from people's pregnant urine.
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| CN114414798A (en) * | 2021-12-08 | 2022-04-29 | 北京泰格科信生物科技有限公司 | Chlamydia trachomatis/gonococcus/mycoplasma genitalium antigen combined detection kit and preparation method thereof |
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Application publication date: 20171024 |