CN107290549A - A kind of kit, its preparation method and detection method for determining anti-Miao Le Shi pipe hormones - Google Patents
A kind of kit, its preparation method and detection method for determining anti-Miao Le Shi pipe hormones Download PDFInfo
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- CN107290549A CN107290549A CN201710568444.9A CN201710568444A CN107290549A CN 107290549 A CN107290549 A CN 107290549A CN 201710568444 A CN201710568444 A CN 201710568444A CN 107290549 A CN107290549 A CN 107290549A
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- 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
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Abstract
A kind of kit, its preparation method and detection method for determining anti-Miao Le Shi pipe hormones, the invention belongs to immune diagnostic technique field, it is an object of the invention to for the deficiencies in the prior art, the present invention uses following technical scheme：Detection band in test strips（T）The anti-AMH antibody corresponding with thing to be checked is coated with, in control band（C）Anti- Avidin antibody is coated with, the anti-AMH antibody of colloid gold label and the Avidin of colloid gold label are coated with conjugates pad.The beneficial effects of the present invention are, present invention detection card can be in Quantitative detection human serum, blood plasma, whole blood AMH concentration, it is simple to operate first, it is easy to use, single part is independently coated with, overcome the drawbacks of small sample amount can not be detected immediately, 20 minutes can reported concentrations result, substantially reduce detection time.
Description
Technical field
The present invention relates to medicine technology field, more particularly to a kind of kit for determining anti-Miao Le Shi pipe hormones and its
Preparation method.
Background technology
Anti- mullerian duct hormone（Anti-Mullerian hormone, AMH）Be transforming growth factor β superfamily into
One of member, was had found first by Professor Alfred Jost in 1974.AMH is, by two identical 70KD subunits, to pass through
The dimerization glycoprotein of disulfide bond composition, relative molecular mass is 140KD；Mankind AMH encoding genes are located at No. 19 chromosomes
Galianconism, 2.4~2.8kb of size contains 5 extrons.
Anti- mullerian duct hormone（AMH）Played an important role in sexual glands growth course, be men and women's gonad function
One of important marker：Mainly produced in male AMH by interstitial glands, start from embry ogenesis and run through life all the time；
In the growth course of male fetus, AMH causes mullerian duct to be degenerated, and forms the male genetic pipeline of normal development.In women AMH
Mainly produced by gonad granulocyte, serum AMH keeps a level relatively low relative to male, from pubarche, serum
AMH levels are slowly reduced with the time, and be reduced to climacteric ELISA method detection less than level.
At present, detect that the Method And Principle used mainly has for AMH both at home and abroad and exempt from ELISA, chemoluminescence method and glue
Body gold method.Chemiluminescence needs specialized analyzer, and expensive, and ELISA is detected using 96 microwell plates mostly, step
Cumbersome time-consuming, colloidal gold method is simple to operate, easy to use, detection speed is fast.
The content of the invention
The general principle that this kit is used is immune colloidal gold chromatography method：Detection band in test strips（T）Be coated with
The corresponding anti-AMH antibody of thing to be checked, in control band（C）Anti- Avidin antibody is coated with, collaurum is coated with conjugates pad
The anti-AMH antibody of mark and the Avidin of colloid gold label.It is to be checked in sample when containing thing to be checked in sample during detection
Thing to be checked on the anti-AMH antibody conjugates formation immune complex of thing and colloid gold label, subsequent immune complex is captured
And it is fixed on one red detection band of formation in nitrocellulose membrane（T）, meanwhile, the Avidin mark logistics on conjugates pad
Through check plot（C）And the anti-Avidin antibody fixed captures to form a red control band, after the completion of reaction, analyzer
Control band and detection band are analyzed, anti-Miao Le Shi pipe hormone concentrations in sample are calculated.
A kind of reagent box preparation method for determining anti-Miao Le Shi pipe hormones, described preparation method comprises the following steps：
First, it is coated with nitrocellulose filter
1）Prepare coating buffer：Anti- AMH antibody and avidin antibody are configured to 0.5-2mg/mL coating concentration respectively；
2）Coating：Nitrocellulose membrane is cut into piece, by step 1）In be coated on respectively on diaphragm with coating buffer, be made anti-containing AMH
The nitrocellulose membrane of body coating buffer and the nitrocellulose membrane of the antibody containing avidin；
3）Dry：The nitrocellulose membrane being coated with is placed in 18-20 hour of 37 DEG C of freeze-day with constant temperature；
2nd, it is coated with gold standard pad
1）Labelled antibody：The mouse monoclonal antibody AMH antibody and avidin conjugates of colloid gold label are prepared, and is configured to 0.5-2mg/mL
Coating concentration；
2）Coating：By gold standard pad dicing, dried after being closed with confining liquid, the coating buffer that previous step is prepared is coated on
In gold standard pad；
3）Dry：The gold standard pad bar being coated with is placed in 18-22 hour of 37 DEG C of freeze-day with constant temperature.
3rd, pad pasting
1st, pasted after combining coated nitrocellulose filter, coating gold standard pad, sample pad and liquid adsorptive pads successively in order
In PVC board, detector bar is made；
4th, cut
1）Will（Three）In the detector bar that posts be cut into the 4.0 ± 0.05mm for cutting into single part band；
5th, assemble
1）Then band is compressed gold and put on lid according to order loaded on gold mark shell.
A kind of application method for the kit for determining anti-Miao Le Shi pipe hormones, comprises the following steps：, kit is kept flat,
Sample to be tested is added in well with sample loading gun, is put into gold-marking immunity analyzer, obtains testing result.
A kind of kit for being used to determine anti-Miao Le Shi pipe hormones.
The beneficial effects of the present invention are present invention detection card can be in Quantitative detection human serum, blood plasma, whole blood
AMH concentration, simple to operate first, easy to use, single part is independently coated with, and overcomes the disadvantage that small sample amount can not be detected immediately
End, 20 minutes can reported concentrations result, substantially reduce detection time.
Embodiment
Below in conjunction with to be illustrated to the present invention：
Embodiment 1
First, it is coated with nitrocellulose filter
1）Prepare coating buffer：Anti- AMH antibody and avidin antibody are configured to 1.5mg/mL coating concentration；
2）Coating：Nitrocellulose membrane is planted into 300mmX25mm diaphragm, by being coated with machine, by 1）The middle coating buffer bag configured
By on diaphragm；
3）Dry：The film strip being coated with is placed in 19 hours of 37 DEG C of freeze-day with constant temperature.
2nd, it is coated with gold standard pad
1）Labelled antibody：Colloid gold label mouse monoclonal antibody AMH antibody and avidin conjugates are prepared, and are configured to 1mg/mL coating
Concentration；
2）Coating：The plain film of glass fibre is cut into 300X13mm diaphragm, dried after being closed with confining liquid, by 1）In prepare
Solution be coated on pad；
3）Dry：The gold standard pad bar being coated with is placed in 20 hours of 37 DEG C of freeze-day with constant temperature.
3rd, pad pasting
1st, dried nitrocellulose filter film, gold standard pad and sample pad, liquid adsorptive pads are pasted according to certain sequential combination
In the front of sticking PVC board；
5th, cut
2）Will（Three）In the diaphragm that posts be cut into the band of one fixed width；
5th, assemble
5.1 slitting：The kilocalorie being laminated is cut into 4.0mm with automatic cutting machine（±0.05mm）Test strips, waste material is all end to end
There should be complete fluorescent microsphere lines.Underproof test strips should be found out having cut by such as lacking in test strips, remove cellulose nitrate
Marked useless bar on plain film；
5.2 put bar：Test strips, which have fluorescent microsphere end to be placed on bottom, has digital one end to be carefully placed into neck, and confirms that each component is complete
It is good；
5.3 covered in pressure：One end alignment of adding mouth is stamped by has fluorescent microsphere end cap is tightly upper to cover, and is put into press flask compression,
And confirm to press closed seamless gap；
5.4 visual inspection：Test strips should be with detecting that parallel, nitrocellulose filter the film surface of card is clean, no breakage, covers whole on detection card
Clean and pattern is complete, bar code is clear, position without skew, sample pad without skew side leakage free, impressed in confirmation brush project name and
Whether production ordering is consistent, and upper and lower covers coordinate closed seamless gap；
Before 5.5 dress drier：Drier is confirmed without breakage, and humidity card (drier is carried) does not have discoloration；Aluminium foil bag is without breakage；
5.6 dress drier：1 drier is put into aluminium foil bag front vertically（It is front to have label surface）The lower left corner；
5.7 dress detection cards：Qualified detection card is put into the right side for having been loaded with 1 drier aluminium foil bag bottom vertically；
Before 5.8 sealings：Sealing temperature sets correct, confirms there is 1 drier and 1 detection card in each aluminium foil bag, and dry
Agent and detection card can not be overlapping, and aluminium foil bag is without breakage；
5.9 sealing：Aluminium foil bag is face-up, sealing machine sealing, and sealing part is answered smooth without twisted；
During detection, detection card is lain against on experimental bench, 100 μ l samples to be tested is separately added into well with sample loading gun, puts
Enter gold-marking immunity analyzer, instrument reports testing result automatically after 20 minutes.
Claims (3)
1. a kind of reagent box preparation method for determining anti-Miao Le Shi pipe hormones, it is characterised in that described preparation method is included such as
Lower step：
First, it is coated with nitrocellulose filter
1）Prepare coating buffer：Anti- AMH antibody and avidin antibody are configured to 0.5-2mg/mL coating concentration respectively；
2）Coating：Nitrocellulose membrane is cut into piece, by step 1）In be coated on respectively on diaphragm with coating buffer, be made anti-containing AMH
The nitrocellulose membrane of body coating buffer and the nitrocellulose membrane of the antibody containing avidin；
3）Dry：The nitrocellulose membrane being coated with is placed in 18-20 hour of 37 DEG C of freeze-day with constant temperature；
2nd, it is coated with gold standard pad
1）Labelled antibody：The mouse monoclonal antibody AMH antibody and avidin conjugates of colloid gold label are prepared, and is configured to 0.5-2mg/mL
Coating concentration；
2）Coating：By gold standard pad dicing, dried after being closed with confining liquid, the coating buffer that previous step is prepared is coated on
In gold standard pad；
3）Dry：The gold standard pad bar being coated with is placed in 18-22 hour of 37 DEG C of freeze-day with constant temperature;
3rd, pad pasting
1st, pasted after combining coated nitrocellulose filter, coating gold standard pad, sample pad and liquid adsorptive pads successively in order
In PVC board, detector bar is made；
4th, cut
1）Will（Three）In the detector bar that posts be cut into the 3.5 ± 0.05mm for cutting into single part band；
5th, assemble
1）Then band is compressed gold and put on lid according to order loaded on gold mark shell.
2. a kind of application method for the kit for determining anti-Miao Le Shi pipe hormones, it is characterised in that comprise the following steps：, will try
Agent box is kept flat, and sample to be tested is added in well with sample loading gun, is put into gold-marking immunity analyzer, is obtained testing result.
3. a kind of kit as claimed in claim 1 or 2 for being used to determine anti-Miao Le Shi pipe hormones.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710568444.9A CN107290549A (en) | 2017-07-13 | 2017-07-13 | A kind of kit, its preparation method and detection method for determining anti-Miao Le Shi pipe hormones |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710568444.9A CN107290549A (en) | 2017-07-13 | 2017-07-13 | A kind of kit, its preparation method and detection method for determining anti-Miao Le Shi pipe hormones |
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| Publication Number | Publication Date |
|---|---|
| CN107290549A true CN107290549A (en) | 2017-10-24 |
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ID=60100747
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710568444.9A Pending CN107290549A (en) | 2017-07-13 | 2017-07-13 | A kind of kit, its preparation method and detection method for determining anti-Miao Le Shi pipe hormones |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110702927A (en) * | 2019-11-04 | 2020-01-17 | 南京欧凯生物科技有限公司 | Colloidal gold immunochromatographic test strip for detecting anti-mullerian hormone and preparation method thereof |
| CN111196851A (en) * | 2018-11-20 | 2020-05-26 | 厦门万泰凯瑞生物技术有限公司 | Specific antibody aiming at human anti-mullerian hormone and application thereof |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102323422A (en) * | 2011-05-30 | 2012-01-18 | 中国科学院上海微系统与信息技术研究所 | Immunochromatographic test strip for semi-quantitatively and simultaneously detecting cTnI and Myo and preparation method thereof |
| WO2013126517A1 (en) * | 2012-02-21 | 2013-08-29 | Northwestern University | Anti-mullerian hormone detection in whole blood |
| CN105181960A (en) * | 2015-10-15 | 2015-12-23 | 厦门宝太生物科技有限公司 | Fluorescent immunochromatography test paper and preparation method thereof |
| CN105527448A (en) * | 2015-12-31 | 2016-04-27 | 苏州市博纳泰科生物技术有限公司 | A fluorescence immunochromatographic detecting method for anti-mullerian hormone and a kit |
| CN105527447A (en) * | 2015-12-10 | 2016-04-27 | 南京沐美生物科技有限公司 | Human anti-mullerian hormone gold immunochromatographic detection kit and method thereof |
| CN105891490A (en) * | 2016-04-05 | 2016-08-24 | 付国亮 | Test strip for quantitatively detecting anti-mullerian hormone, preparation method thereof and determination method for concentration of anti-mullerian hormone |
| CN106290832A (en) * | 2016-08-12 | 2017-01-04 | 上海铭源数康生物芯片有限公司 | A kind of immunity lateral chromatography quantitative detecting reagent and preparation method thereof, detection method |
| CN206038685U (en) * | 2016-07-06 | 2017-03-22 | 北京康思润业生物技术有限公司 | Immunity side direction chromatography detecting system |
-
2017
- 2017-07-13 CN CN201710568444.9A patent/CN107290549A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102323422A (en) * | 2011-05-30 | 2012-01-18 | 中国科学院上海微系统与信息技术研究所 | Immunochromatographic test strip for semi-quantitatively and simultaneously detecting cTnI and Myo and preparation method thereof |
| WO2013126517A1 (en) * | 2012-02-21 | 2013-08-29 | Northwestern University | Anti-mullerian hormone detection in whole blood |
| CN105181960A (en) * | 2015-10-15 | 2015-12-23 | 厦门宝太生物科技有限公司 | Fluorescent immunochromatography test paper and preparation method thereof |
| CN105527447A (en) * | 2015-12-10 | 2016-04-27 | 南京沐美生物科技有限公司 | Human anti-mullerian hormone gold immunochromatographic detection kit and method thereof |
| CN105527448A (en) * | 2015-12-31 | 2016-04-27 | 苏州市博纳泰科生物技术有限公司 | A fluorescence immunochromatographic detecting method for anti-mullerian hormone and a kit |
| CN105891490A (en) * | 2016-04-05 | 2016-08-24 | 付国亮 | Test strip for quantitatively detecting anti-mullerian hormone, preparation method thereof and determination method for concentration of anti-mullerian hormone |
| CN206038685U (en) * | 2016-07-06 | 2017-03-22 | 北京康思润业生物技术有限公司 | Immunity side direction chromatography detecting system |
| CN106290832A (en) * | 2016-08-12 | 2017-01-04 | 上海铭源数康生物芯片有限公司 | A kind of immunity lateral chromatography quantitative detecting reagent and preparation method thereof, detection method |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111196851A (en) * | 2018-11-20 | 2020-05-26 | 厦门万泰凯瑞生物技术有限公司 | Specific antibody aiming at human anti-mullerian hormone and application thereof |
| CN111196851B (en) * | 2018-11-20 | 2021-11-16 | 厦门万泰凯瑞生物技术有限公司 | Specific antibody aiming at human anti-mullerian hormone and application thereof |
| CN110702927A (en) * | 2019-11-04 | 2020-01-17 | 南京欧凯生物科技有限公司 | Colloidal gold immunochromatographic test strip for detecting anti-mullerian hormone and preparation method thereof |
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Application publication date: 20171024 |
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