CN109406773A - A kind of capillary detection kit and preparation method thereof for interstitialcellstimulating hormone (ICSH) - Google Patents
A kind of capillary detection kit and preparation method thereof for interstitialcellstimulating hormone (ICSH) Download PDFInfo
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- CN109406773A CN109406773A CN201811614753.6A CN201811614753A CN109406773A CN 109406773 A CN109406773 A CN 109406773A CN 201811614753 A CN201811614753 A CN 201811614753A CN 109406773 A CN109406773 A CN 109406773A
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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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
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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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
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Abstract
The capillary detection kit and preparation method thereof that the present invention provides a kind of for interstitialcellstimulating hormone (ICSH), comprising: capillary detection pipe, blotting paper, drop bottle and probe tube；The capillary detection pipe is the capillary that surface is from top to bottom provided with nature controlling line and max line, nature controlling line area above is quality control region in the capillary detection pipe, max line following region is pattern detection area, the inner wall in the pattern detection area is coated with first antibody, and the inner wall of the quality control region is coated with the secondary antibody of secondary antibody；The secondary antibody of color latex microballoon label is loaded in the drop bottle；The first antibody and secondary antibody are interstitialcellstimulating hormone (ICSH) antibody.Kit of the present invention simplifies experimental implementation process, reduces using difficulty, improves product quality while simplification using tool, while reducing process costs.
Description
Technical field
The invention belongs to medical instruments fields, more particularly, to a kind of capillary detection reagent for interstitialcellstimulating hormone (ICSH)
Box and preparation method thereof.
Background technique
Interstitialcellstimulating hormone (ICSH) (LH) is the substance secreted by adenohypophysis basophil.It is common in women (LH) collaboration FSH
Effect maintains the menstrual cycle of ovary, and ovulation is caused to be formed with corpus luteum.Control of the generation of LH by hypothalamus gonadotropin releasing hormone
System, while being regulated and controled by the positive-negative feedback of ovary.On the release peak of menstrual cycle LH and ovary ovulation close relation, LH
Peak indicates ovary ovulation in 24-36 hours once appearance, therefore serum Lh peak value can be monitored in the menstrual cycle, with determination
Best Time to pregnancy.The testing result is indicated with milli-international unit/milliliter (MIU/ML).
Family common are card-type and two kinds of pen type, block for being pregnant testing product based on colloidal gold colloidal gold detection test paper strip
Type needs the pipettor or rubber head dropper of profession, and pen type, which can help a small child urinate by holding his legs apart directly to pour, pours detection zone, and two kinds of reagent costs are opposite
Higher, technique is also relatively complicated, and still, its detection speed ratio is very fast, and accuracy different manufacturers are slightly different, the amount of convenience
It produces.
Capillary phenomenon (also known as capillarity) refers to liquid on the inside of thin tube-like object, due to cohesive force and adhesive force
Difference, the phenomenon that overcoming centrifugal force and rise or fall.When object containing micropore and liquid contact, make the liquid
The phenomenon that being risen or fallen along hole.When the adhesive force between liquid and solid (tube wall) is greater than liquid cohesive force itself, just
Capillary phenomenon (rising) can be generated；Conversely, when the adhesive force between liquid and solid (tube wall) is less than liquid cohesive force itself,
Capillary phenomenon (decline) will be generated.Liquid level is in shape concave or convex and porous material object energy when liquid is in vertical tubule
Absorbing liquid is all caused by this phenomenon.The appearance of capillarity is that due to water there is stickiness-hydrone to stick together mutually to be attached to it
Characteristic on his object, these objects can be glass, cloth, organ-tissue or soil.And mercury is because of the cohesion between its atom
Power is extremely strong, so capillary phenomenon (decline) occurs.Effects of air pressure suffered by thinner capillary suction is more unobvious, so thinner
Capillary perpendicular to the water surface water sucting degree it is stronger.
There are following performance characteristics for color latex microballoon: (1) sensitivity is higher, the ideal substitute of colloidal gold；(2) internal
Dyeing, color is full securely, no dyestuff surface, convenient for coupling；(3) water-wetted surface and group content are abundant, so that albumen
Binding ability is higher；(4) scale supply and performance stabilization can be achieved, it can be achieved that single batch highest 100L yield；(5) framboid
Diameter is uniform, and conventional particle size can realize C.V. less than 5%；(6) customizable produces, particle size (100nm~10um), surface base
Mass contg and rainbow series color are adjustable.
There is not the related report that capillarity is used for interstitialcellstimulating hormone (ICSH) detection in conjunction with color latex microballoon also at present
Road and document.
Summary of the invention
In view of this, the present invention is directed to propose a kind of for the capillary detection kit of interstitialcellstimulating hormone (ICSH) and its preparation
Method, the kit simplify experimental implementation process, reduce using difficulty, improve product matter while simplification using tool
Amount, while reducing process costs.
In order to achieve the above objectives, the technical scheme of the present invention is realized as follows:
A kind of capillary detection kit for interstitialcellstimulating hormone (ICSH), comprising: capillary detection pipe, blotting paper, drop bottle and
Probe tube；
The capillary detection pipe is the capillary that surface is from top to bottom provided with nature controlling line and max line, the capillary detection
Nature controlling line area above is quality control region in pipe, and max line following region is pattern detection area, and the inner wall in the pattern detection area is coated with
There is first antibody, the inner wall of the quality control region is coated with the secondary antibody of secondary antibody；
The secondary antibody of color latex microballoon label is loaded in the drop bottle；
The first antibody and secondary antibody are interstitialcellstimulating hormone (ICSH) antibody.
Further, the secondary antibody of the first antibody, the secondary antibody of secondary antibody and color latex microballoon label is equal
It is diluted by buffer.
Further, the buffer is 0.01M PBS.
Further, the concentration of the first antibody after buffer dilution is 0.1-1mg/ml；Second after buffer dilution
The concentration of the secondary antibody of antibody is 0.1-1mg/ml, the volume hundred of the secondary antibody of the color latex microballoon label after buffer dilution
Divide than being 1-10%.
Further, the probe tube is eight connecting legs.
Further, the kit further includes rubber head compatible with capillary detection pipe.
The preparation method of the above-mentioned capillary detection kit for interstitialcellstimulating hormone (ICSH), includes the following steps:
S1: capillary is from top to bottom identified to nature controlling line and max line, wherein max line is attainable for sample upstream end
Maximum value, for max line the following are pattern detection area, nature controlling line is that Quality Control end reaches maximum value, nature controlling line and the above are quality control regions；
S2: being diluted with buffer and pass through coated first antibody, is encapsulated spare；With the two of buffer dilution secondary antibody
It is anti-, it encapsulates spare；The secondary antibody marked with buffer dilution color latex microballoon, and seal, be kept in dark place in drop bottle；
S3: the sample end of capillary is inserted into the first antibody solution diluted, until liquid level rises to max line,
It takes out and dries；
S4: the Quality Control end of capillary is inserted into the secondary antibody diluent of secondary antibody, until liquid level rises to nature controlling line, is taken
Out and dry to get capillary detection pipe；
S5: the capillary detection pipe prepared is put into desiccant seal up for safekeeping it is spare；
S6: the capillary detection pipe sealed up for safekeeping, drop bottle, blotting paper, eight connecting legs, the rubber head for being adapted to capillary and desiccant are sealed up for safekeeping
The capillary detection kit of interstitialcellstimulating hormone (ICSH) is used for after in container.
Further, the temperature dried in the step S3 is 45 DEG C, time 1h.
Further, the temperature dried in the step S4 is 45 DEG C and stays overnight.
Mentioned reagent box applies also for the detection of other albumen, antibody or antigen.
Compared with the existing technology, of the present invention for the capillary detection kit of interstitialcellstimulating hormone (ICSH) and its preparation
Method has the advantage that
(1) kit of the present invention is convenient to use does not need professional tool；It is inhaled automatically using capillary effect
Liquid is taken, carrying simple auxiliary detection instrument, (wherein, rubber head prevents liquid level not to be able to satisfy experiment demand；Eight connecting legs are for examining
Test agent is held, and prevents cross contamination；Blotting paper is for adsorbing useless reagent and sample)；
It (2) is main detection device using capillary in kit of the present invention, compared to colloidal gold and latex
The cost of product is low；
(3) user of service of kit of the present invention does not need professional guidance, using color latex microballoon, develops the color bright
It is aobvious, easy interpretation；
(4) reagent stability used in kit of the present invention is good, so it is low to environmental requirement；
(5) seminal plasma fructose detection kit of the present invention is pollution-free, and experiment rubbish is few and small, and environmental protection is easy to handle；
(6) kit of the present invention can do more people's detections simultaneously, and the time difference is extremely short, and high-volume is facilitated to screen,
It is easy to use；
(7) kit of the present invention is suitable for multiple protein, antigen, antibody test, is particularly suitable for liquid sample class
Type detection.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of capillary detection pipe in kit of the present invention.
Appended drawing reference:
1, Quality Control end；2, quality control region；3, nature controlling line；4, max line；5, pattern detection area；6, sample upstream end.
Specific embodiment
In addition to being defined, technical term used in following embodiment has universal with those skilled in the art of the invention
The identical meanings of understanding.Test reagent used in following embodiment is unless otherwise specified conventional biochemical reagent；It is described
Experimental method is unless otherwise specified conventional method.
Below with reference to examples and drawings, the present invention will be described in detail.
A kind of capillary detection kit for interstitialcellstimulating hormone (ICSH), comprising: capillary detection pipe, blotting paper, drop bottle, glue
Head and probe tube；
Capillary detection pipe, as shown in Figure 1, being from top to bottom provided with the capillary of nature controlling line 3 and max line 4, capillary for surface
Tubing matter is polystyrene, and 3 area above of nature controlling line is quality control region 2, and 4 following region of max line is pattern detection area 5, sample inspection
The inner wall for surveying area 5 is coated with first antibody, and the inner wall of quality control region 2 is coated with the secondary antibody of secondary antibody；
The secondary antibody of color micro-sphere label is loaded in drop bottle；
Specifically, probe tube is eight connecting legs, in order to hold detection reagent.
Specifically, rubber head is adapted with capillary detection pipe, and liquid level is not able to satisfy experiment demand in order to prevent.
Specifically, blotting paper is for adsorbing useless reagent and sample.
Wherein, first antibody and secondary antibody are interstitialcellstimulating hormone (ICSH) antibody.
Specifically, the secondary antibody of first antibody, the secondary antibody of secondary antibody and color latex microballoon label is through too slow
Fliud flushing dilution.Buffer uses 0.01M PBS.
Specifically, the concentration of the first antibody after buffer dilution is 1mg/ml；Secondary antibody after buffer dilution
The concentration of secondary antibody is 1mg/ml, and the percent by volume of the secondary antibody of the color latex microballoon label after buffer dilution is 5%.
Kit of the invention the preparation method is as follows:
S1: capillary is from top to bottom identified to nature controlling line and max line, wherein max line is attainable for sample upstream end
Maximum value, for max line the following are pattern detection area, nature controlling line is that Quality Control end reaches maximum value, nature controlling line and the above are quality control regions；
S2: first antibody is diluted to respective concentration with buffer, encapsulation is standby to deposit；With the secondary antibody of buffer dilution secondary antibody
To respective concentration, encapsulation is standby to be deposited；With buffer dilution color latex microballoon label secondary antibody, and seal, be kept in dark place in
In drop bottle；
S3: the sample insertion end of capillary is inserted into the first antibody solution diluted, until liquid level rises to max
Line is removed and placed in 1h in 45 DEG C of baking ovens；
S4: the Quality Control end of capillary is inserted into the secondary antibody diluent of secondary antibody, until liquid level rises to nature controlling line, is taken
Out and it is put into baking oven, 45 DEG C overnight to get capillary detection pipe；
S5: the capillary detection pipe prepared is put into desiccant seal up for safekeeping it is spare；
S6: the capillary detection pipe sealed up for safekeeping, drop bottle, blotting paper, eight connecting legs, the rubber head for being adapted to capillary and desiccant are sealed up for safekeeping
The capillary detection kit of interstitialcellstimulating hormone (ICSH) is used for after in container.
The application method of mentioned reagent box are as follows: (secondary antibody of the first antibody of LH, secondary antibody and secondary antibody, respectively
It is indicated with Ab1, Ab2, secondary antibody)
A. capillary detection pipe, blotting paper, dropper and eight connecting legs are taken out, the sample upstream end of capillary detection pipe is inserted into liquid
In sample, through capillary action, sample can be adsorbed therewith increases to max line or less area, capillary detection pipe is taken out, if sample
Containing Ag to be detected (english abbreviation of antigen), that is, the compound that will form Ab1-Ag is adsorbed on capillary tube inner wall, if
This compound is not will form without Ag.
B. sample is emptied by the sample upstream end 6 of capillary detection pipe, i.e., it will be in capillary detection pipe with dry blotting paper
Liquid sample is sucked out, and still can be adsorbed on inner wall if forming compound.
C. eight connecting legs are vertically put on the table, the Ab2 examination of color latex microballoon label is added dropwise into eight connecting legs with drop bottle
Agent is inserted into the sample upstream end 6 of capillary detection pipe in the Ab2 reagent of color latex microballoon label, Ab2- color latex microballoon
It can go up with capillarity to top, if containing Ag to be detected in sample, in pattern detection area, i.e. max line portion below
Position, just will form the compound of Ab1-Ag-Ab2- color latex microballoon, is adsorbed on inner wall, the Ab2 of color latex microballoon label
Reagent will continue to go up to Quality Control region, and two anti-bindings with the Ab2 antibody being coated in advance form the colored cream of two anti-Ab2-
The compound of glue microballoon is adsorbed in the capillary tube inner wall of quality control region.
D. reagent (dry blotting paper can be sucked out) is discharged by the Quality Control end 1 of capillary detection pipe.
Result judgement:
If being free of Ag in sample, sample detection zone will not form coloured complex Ab1-Ag-Ab2- color latex microballoon,
So capillary detection pipe color change cannot be observed from appearance, as negative, color latex microballoon can be anti-with the Ab2 of quality control region
Two anti-bindings of body form the coloured complex of two anti-Ab2- color latex microballoons, so quality control region has color change；
If containing Ag to be detected in sample, sample detection zone will form compound Ab1-Ag-Ab2- color micro-sphere, institute
It can observe capillary detection pipe color change with appearance, i.e., it is positive, while as described above, quality control region also has colour switching.
In summary: sample no matter is detected to be negative or positive, quality control region can all change colour, and it is non-discolouring, prove that reagent goes out
Existing problem, cannot use；Then detection zone is colourless, quality control region discoloration to negative sample；Positive sample then quality control region and detection
Area changes colour simultaneously.
The invention advantage is as follows: first, corresponding product mainly has colloidal gold and emulsion products, it is all quickly to examine
Disconnected favorite, colloidal gold and emulsion products detection speed are fast, using relatively convenient, still, and for conventional kitchen, a part
Test instrument does not have, such as pipettor etc., and kit detection of the invention does not need special experimental facilities, passes through capillary
Action draws, two kinds of experiment types, the present invention are lower to equipment requirement relatively above.Second, the two of colloidal gold and emulsion products
Kind product needs the personnel of profession to teach the judgement of detection zone in the detection process, and some weakly positive poor signals judge, and
The sensitivity for this reagent that the present invention designs is higher to keep color difference more obvious, easily judges.Third, both the above method prepares work
Skill is relative complex, needs a large amount of matched instrument and equipment, and the detection of method name does not need complicated instrument and equipment, technique letter
It is single, low manufacture cost.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Within mind and principle, any modification, equivalent replacement, improvement and so on be should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of capillary detection kit for interstitialcellstimulating hormone (ICSH), it is characterised in that: include: capillary detection pipe, water suction
Paper, drop bottle and probe tube；
The capillary detection pipe is the capillary that surface is from top to bottom disposed with nature controlling line and max line, the capillary detection
Nature controlling line area above is quality control region in pipe, and max line following region is pattern detection area, and the inner wall in the pattern detection area is coated with
There is first antibody, the inner wall of the quality control region is coated with the secondary antibody of secondary antibody；
The secondary antibody of color latex microballoon label is loaded in the drop bottle；
The first antibody and secondary antibody are the antibody of interstitialcellstimulating hormone (ICSH).
2. the capillary detection kit according to claim 1 for interstitialcellstimulating hormone (ICSH), it is characterised in that: described
The secondary antibody of one antibody, the secondary antibody of secondary antibody and color latex microballoon label passes through buffer dilution.
3. the capillary detection kit according to claim 2 for interstitialcellstimulating hormone (ICSH), it is characterised in that: buffer
The concentration of first antibody after dilution is 0.1-1mg/ml；The concentration of the secondary antibody of secondary antibody after buffer dilution is 0.1-
The percent by volume of 1mg/ml, the secondary antibody of the color latex microballoon label after buffer dilution are 1-10%.
4. the capillary detection kit according to claim 3 for interstitialcellstimulating hormone (ICSH), it is characterised in that: described
Buffer is 0.01M PBS.
5. the capillary detection kit according to claim 1 for interstitialcellstimulating hormone (ICSH), it is characterised in that: described to take
Sample pipe is eight connecting legs.
6. the capillary detection kit according to claim 1 for interstitialcellstimulating hormone (ICSH), it is characterised in that: the examination
Agent box further includes rubber head compatible with capillary detection pipe.
7. the preparation method of any capillary detection kit for interstitialcellstimulating hormone (ICSH) of claim 1-6, special
Sign is: including the following steps:
S1: capillary is from top to bottom identified to nature controlling line and max line, wherein max line is the attainable maximum of sample upstream end
Value, for max line the following are pattern detection area, nature controlling line is that Quality Control end reaches maximum value, nature controlling line and the above are quality control regions；
S2: first antibody is diluted with buffer, is encapsulated spare；With the secondary antibody of buffer dilution secondary antibody, encapsulate spare；With slow
Fliud flushing dilutes color latex microballoon and marks secondary antibody, and seals, is kept in dark place in drop bottle；
S3: the sample end of capillary is inserted into the first antibody solution diluted, and until liquid level rises to max, taking-up is simultaneously
Drying；
S4: the Quality Control end of capillary is inserted into the secondary antibody diluent of secondary antibody, and until liquid level rises to nature controlling line, taking-up is simultaneously
Drying is to get capillary detection pipe；
S5: it is sealed up for safekeeping after the capillary detection pipe and desiccant that prepare are stored jointly spare；
S6: the capillary detection pipe sealed up for safekeeping, drop bottle, blotting paper, eight connecting legs, the rubber head for being adapted to capillary and desiccant are sealed in appearance
The capillary detection kit of interstitialcellstimulating hormone (ICSH) is used for after in device.
8. the preparation method of the capillary detection kit according to claim 7 for interstitialcellstimulating hormone (ICSH), feature
Be: the temperature dried in the step S3 is 45 DEG C, time 1h.
9. the preparation method of the capillary detection kit according to claim 7 for interstitialcellstimulating hormone (ICSH), feature
Be: the temperature dried in the step S4 is 45 DEG C and stays overnight.
10. aforementioned described in any item kits are detecting the application in other albumen, antibody or antigen.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811614753.6A CN109406773A (en) | 2018-12-27 | 2018-12-27 | A kind of capillary detection kit and preparation method thereof for interstitialcellstimulating hormone (ICSH) |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811614753.6A CN109406773A (en) | 2018-12-27 | 2018-12-27 | A kind of capillary detection kit and preparation method thereof for interstitialcellstimulating hormone (ICSH) |
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| CN109406773A true CN109406773A (en) | 2019-03-01 |
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