CN106370849B - Immune lateral chromatography detection system and preparation method thereof - Google Patents
Immune lateral chromatography detection system and preparation method thereof Download PDFInfo
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- CN106370849B CN106370849B CN201610921295.5A CN201610921295A CN106370849B CN 106370849 B CN106370849 B CN 106370849B CN 201610921295 A CN201610921295 A CN 201610921295A CN 106370849 B CN106370849 B CN 106370849B
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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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- 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
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Abstract
The invention discloses a kind of immune lateral chromatography detection systems and preparation method thereof, and wherein detection system includes: bottom plate comprising for carrying the first offset plate of sample pad, nitrocellulose filter and the second offset plate for carrying water absorption pad;First offset plate and the second offset plate is fixedly connected with the both ends of the nitrocellulose filter respectively;Detection band and quality control band are provided on the nitrocellulose filter;Sample pad is fixed on first offset plate and contacts with the nitrocellulose filter to guarantee to climb water;Water absorption pad is fixed on second offset plate and contacts with the nitrocellulose filter to guarantee to climb water.Detection system of the invention has the advantages that at low cost, to climb water effect good.
Description
Technical field
The present invention relates to medical domains, and in particular to a kind of immune lateral chromatography detection system and preparation method thereof.
Background technique
Immune lateral chromatography diagnostic techniques is suitble to the real-time test in multiplicity as a kind of stabilization and practical technology
(POCT) or scene uses.Being divided into mainly by mark part has colloid gold immune lateral chromatography method, common fluorescent immune lateral
Chromatography, time-resolved fluoroimmunoassay lateral chromatography method, on turn electrochemiluminescent immunoassay lateral chromatography method, lateral layer is immunized in quantum dot fluorescence
Analysis method etc. mainly has nitrocellulose filter, composite material (fusion5), micro-fluidic etc. by coating part.
With the development of immunochromatography technique (immunochromatography) and colloidal gold technique, the especially nineties
Afterwards, diagnose the illness colloid gold immune lateral chromatography method (Gold immunochromatography assay, GICA) inspection in vitro
It is widely applied in survey.Quantitative detection is but carried out now, requires project precision and repeatability higher and higher, process
The development in many years, nearest all kinds of fluorescence immunoassay lateral chromatography methods emerge one after another.Mainstream advanced technology is had become by many public affairs
Department praises highly.
Current fluorescence immunoassay lateral chromatography method is generally used as using nitrocellulose filter and climbs water material, however at present
The length of nitrocellulose filter that uses of detection device it is longer, it is bad to climb water effect, and offset plate is when assemble reagent card, need
Nitrocellulose filter, sample pad, water absorption pad are first cut (5mm wide) then recomposition, packaging efficiency is low, and technique is cumbersome, because
Sample pad has processed, the sequence of sample pad both forward and reverse directions, front-rear direction and sample pad be all it is random, there are also manually putting
The precision when error of the position of sample pad all can influence finally to detect.
Summary of the invention
The purpose of the present invention is to provide a kind of immune lateral chromatography detection systems, can effectively reduce nitrocellulose filter
Length, reduce detection time, improve the accuracy of detection, saved cost.
The present invention also provides a kind of preparation methods of immune lateral chromatography detection system.
The purpose of the present invention is what is be achieved through the following technical solutions:
The present invention provides a kind of immune lateral chromatography detection systems, comprising:
Bottom plate comprising for carrying the first offset plate of sample pad, nitrocellulose filter and for carrying water absorption pad
Two offset plates;First offset plate and the second offset plate is fixedly connected with the both ends of the nitrocellulose filter respectively;Described
Detection band and quality control band are provided on nitrocellulose filter;
Sample pad is fixed on first offset plate and contacts with the nitrocellulose filter to guarantee to climb water;
Water absorption pad is fixed on second offset plate and contacts with the nitrocellulose filter to guarantee to climb water.
It further, further include card plug comprising:
Kerve is used to fix the bottom plate;
Upper cover is provided with the well for being loaded in the sample pad;
Window is detected, is used to detect the detection window of the detection band and quality control band;The detection window is arranged in kerve
Or on upper lid.
Further, the baseplate width is 4-6mm, length 6.5-7.5cm.
Further, the length of first offset plate is 1.8-2.0cm;The length of second offset plate is 1.5-
1.7cm;The length of the sample pad is 1.7-1.8cm;The length of the water absorption pad is 1.2-1.4cm.
The present invention also provides a kind of preparation methods of immune lateral chromatography detection system, include the following steps:
(1) the coating detection band and quality control band on nitrocellulose filter, by the two sides of nitrocellulose filter respectively with offset plate A
It is fixedly connected with offset plate B, the detection band is between offset plate A and quality control band;
(2) sample pad is fixed on the offset plate A and is contacted with the nitrocellulose filter to guarantee to climb water, it will
Water absorption pad is fixed on the offset plate B and contacts with the nitrocellulose filter to guarantee to climb water;Obtain detection system hair
Base;
(3) the detection system blank is cut into required size up to the immune lateral chromatography detection system.
Further, the detection system blank is cut into after required size further includes following steps:
It is put into card plug after the detection system blank is cut into required size, the card plug includes:
Kerve is used to fix the bottom plate;The kerve bottom is equipped with for detecting the detection band and Quality Control
The detection window of band;
Upper cover is provided with the well for being loaded in the sample pad.
Further, the step (2) specifically comprises the following steps:
With the fixed bottom plate of " work " pattern tool, the sample pad is pasted on offset plate A and pushes down the nitre
Acid cellulose film, pushing down width is 1-3mm;The water absorption pad is pasted on offset plate B and pushes down the nitrocellulose
Film, pushing down width is 1-3mm.
Further, the offset plate A width is 1.8-2.0cm, and the width of the nitrocellulose filter is 3.3-
The width of 3.8cm, the offset plate B are 1.5-1.7cm.
Further, it is described it is required having a size of width be 4-6mm.
Compared with prior art, the present patent application at least have it is following the utility model has the advantages that
It will be arranged to offset plate below sample pad and water absorption pad, the length for reducing nitrocellulose filter (is reduced to original
Carry out the half or so of length), since the surface of offset plate is similar to mirror surface, to the resistance very little for climbing water, sample pad and suction
Water cushion aperture is very big, to the resistance very little for climbing water, compared with nitrocellulose filter, it might even be possible to ignore, this is just dropped significantly
It is low to climb the distance of water, the water time is climbed to reduce, improves the accuracy of testing result.After wedge-shaped card adds offset plate, than
In the case that product overall length is constant, nitrocellulose film length reduces to 3.5cm, nitrocellulose filter cost by original 7cm
It is 10 times or so of offset plate, cost of material can be reduced.
Wedge-shaped card, when assembling reagent card, needs that nitrocellulose filter, sample pad, water absorption pad are first cut (5mm without offset plate
It is wide) then recomposition, packaging efficiency is low, and technique is cumbersome;And after offset plate is added in new system, it can be when assembling kilocalorie (before cutting out)
Sample pad, water absorption pad are posted, then cut again, packaging efficiency greatly improves, and can reduce cost of labor, because of sample pad
It is processed, the sequence of sample pad both forward and reverse directions, front-rear direction and sample pad be all it is random, there are also the positions of artificial setting-out product pad
The precision when error set all can influence finally to detect;And after offset plate is added in new system, it can be in kilocalorie sample pad, suction
Water cushion posts, and then cuts again, sample pad both forward and reverse directions, in the front-back direction and the sequence of sample pad is determined, and makes simultaneously
The location error of sample pad is smaller, reduces detection CV, improves system precision and repeatability.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of existing detection card in the immune lateral chromatography detection system embodiment of the present invention;
Fig. 2 is a kind of structural schematic diagram of detection card in the immune lateral chromatography detection system embodiment of the present invention;
Fig. 3 is a kind of schematic diagram of internal structure of kerve in the immune lateral chromatography detection system embodiment of the present invention;
Fig. 4 is a kind of structural schematic diagram of upper cover in the immune lateral chromatography detection system embodiment of the present invention.
Specific embodiment
For convenience of those skilled in the art understand that technical solution of the present invention, with reference to the accompanying drawing and preferred embodiment is to this hair
Bright technical solution is further elaborated, it should be understood that preferred embodiment is the understanding of this programme for convenience, is not intended as this
The restriction of invention.
The detection card of existing immune lateral chromatography system is as shown in Figure 1 comprising nitrocellulose filter 1,2 He of sample pad
Water absorption pad 3, sample pad 2 and water absorption pad 3 are separately positioned on 1 both ends of nitrocellulose filter, and sample pad 2 and water absorption pad 3 are all entire
It is pressed on nitrocellulose filter 1, climb water by the capillarity of nitrocellulose filter 1, on nitrocellulose filter 1 successively
It is provided with detection band 4 and quality control band 5, is respectively used to detect and verify testing result.
The immune lateral chromatography detection system of the present invention program it is as shown in Figure 2 comprising nitrocellulose filter 8, nitric acid
Cellulose membrane both ends are connect with offset plate, and wherein one end is the first offset plate 6 for carrying sample pad 2, and the other end is for holding
The second offset plate 7 of water absorption pad 3 is carried, sample pad 2 and water absorption pad 3 are separately fixed on corresponding offset plate, on nitrocellulose filter 8
It is provided with detection band 4 and quality control band 5, is respectively used to detect and verify testing result.
Compared with existing detection card, detection card both ends of the invention replace part nitrocellulose filter using offset plate,
Shortening is climbed the water time, and the cost of detection card is reduced.
It will be clear that the present invention program to detection band 4 and quality control band 5 sequence of positions without limitation, according to need
Select suitable sequence and position.
Embodiment 1
Immune lateral chromatography detection system provided by the invention the preparation method is as follows:
Stick lower offset plate:
Nitrocellulose filter specification: 23*3.5cm,
Offset plate A specification: 23* (1.9+0.2) cm, having at 2mm in side has a tangent line;
Offset plate B specification: 23* (1.6+0.2) cm, having at 2mm in side has a tangent line;
Note: offset plate surface has double-sided adhesive, and the tangent line of two sides is to cut double-sided adhesive, facilitates subsequent and nitrocellulose filter
Connection.
Two blocks of offset plates are attached to nitre by paster outside the double-sided adhesive of the 2mm wide to be torn on the outside of two pieces of offset plate tangent lines respectively respectively
The two sides of acid cellulose film, that does not tear double-sided adhesive is partially located in nitrocellulose filter outside, and the part torn is pressed in nitric acid fibre
It ties up on plain film.
Paste sample pad:
The nitrocellulose filter of offset plate will be posted, according to the locality indicated, water absorption pad end is put into and is made upper
In " I " fonts mold.Upper edge and the mold alignment for ensuring backboard, avoid leaving gap, push down nitric acid fibre with " I " fonts fixture
Tie up plain film and by its clamping in a mold (one side of non cohesive gel plate is upper), then tear sample pad location and (remove 2mm on offset plate A
Part other than wide small item) double-sided adhesive (specification: 23*1.9cm).Be careful not to the water absorption pad of the mold of " I " fonts and
Sample pad end is put back.
By the sample pad cut (wide 1.75cm), along the close coated film side of " I " fonts mold since one end pair
(sample pad pushes down nitrocellulose filter 2mm) together, sample pad is successively pressed lightly on to the other end and is attached on offset plate.
The double-sided adhesive paster torn before using again is gently placed in the sample pad posted, and right-handed index finger is again from a left side
It presses one time to the right, it is ensured that then double-sided adhesive paster is taken down in the smooth and stickup completely of sample pad.
By the water absorption pad cut (wide 1.3cm) according to patch sample pad the step of be attached on offset plate B, by the profiled sheeting posted from
It is taken out in mold.The extra sample pad in profiled sheeting both sides and water absorption pad are cut with scissors.
Slitting
The above-mentioned big plate posted is cut into required width detection card, generally 5mm width.Detect the structural representation of card
Scheme as shown in Figure 2 (structure is for example aforementioned).
Group card
Plastics card plug introduction
Plastics card plug is made of upper cover and bottom cover two parts, and Fig. 3 is the structural schematic diagram of kerve, and Fig. 4 is the structure of upper cover
Schematic diagram;As shown in Fig. 2, kerve includes fixation hole 11, it is used to fix kerve and upper cover, detects card placing groove, by solid
Fixed board 9 and the limiting slot at both ends 10 surround, for placing detection card;Middle part and nitrocellulose filter 8 in detection card placing groove
Corresponding position is provided with detection window 12, is used to collect the testing result of detection band 4 and quality control band 5.
Slot bottom is confirmed before being loaded without dirty, completely without incompleteness, the fixed plate 9 of especially detection card placing groove two sides is complete,
It can be used normally.
Confirmation detection card is complete, sample pad, water absorption pad N/D, detection card specification shape, film no marking, hairless invariably
Side, film surface are without signature pen trace etc..
By the site at the top of the card plug of water absorption pad one end alignment card bottom suction side, card compacting then will test, it is ensured that detection
Block all smooth be placed in film slot and the contact surface of slot bottom is seamless, no shaking.
Buckle covers beyond the Great Wall:
Cap sheathing mechanism is used cooperatively with by upper cover as shown in figure 4, there is fixed column 13 in inside with the fixation hole 11 on kerve
It is fixed together with kerve;Sample holes 14 are provided at position corresponding with sample pad 2, are used to that sample to be added dropwise in sample pad 2
Product.
Check whether upper cover is complete, whether has sprayed bar code on upper lid, whether there is or not damaged or incomplete, it is ensured that upper cover is intact, can be with
Normal use.
Be placed in assembled kerve is neat on levels operation table top, and check whether the position of detection card accurate,
Completely, test strips sequence not put back.
Upper cover is buckled on kerve (sample holes 14 of upper cover and 2 extreme direction of sample pad of kerve are consistent) again, it will be upper with hand
Lid is pushed down.
It by assembled detection system, is numbered according to nitrocellulose filter, is put on case pressing machine respectively and carries out pressure shell.It presses
Shell to be checked, check whether the upper and lower covers of card plug press sternly.
Detection system that is assembled, posting label is successively placed in the valve bag accordingly numbered, is packed into appropriate dry
Agent places into aluminium foil bag, and carries out mark, seals sack with sealing machine, spare.
Embodiment 2
A kind of preparation method of immune lateral chromatography detection system, includes the following steps:
Coated antibody: human muscle hemoglobin (MYO) detection antibody 1 is diluted to fixed concentration (2.0mg/ with coating buffer
Ml), contrast agents 1 (goat-anti chicken IgY) are diluted to fixed concentration (2.0mg/ml), using special coating equipment BIODOT company
XYZ3060 above-mentioned two liquid is coated on Sai Duolisi nitrocellulose filter 140 (NC film), 37 DEG C of drying boxes dry 4
Hour (respectively obtains detection band and quality control band), spare.It is coated with the phosphate buffer (PBs) plus 3% that buffer is 0.01Mol/l
Sucrose as protective agent.
Labelled antibody: detecting antibody 2 and contrast agents 2 (chicken IgY) latex fluorescent marker for human muscle hemoglobin (MYO),
It is stored in storing liquid, spare, (50mMol/l Tris, 0.5%BSA, pH7.8).
Marker prepares: above-mentioned labelled antibody is dried standby by 5 microlitres of volumes of required concentration specking in testing tube
With.
Sample pad preparation: by sample treatment liquid specking in or on sample pad, drying for standby.Sample treatment liquid are as follows: wherein
Tris-CL is molar concentration, remaining is mass percentage.Specific formula is as follows: Tris-CL:50mMol/l, and 0.5%
Casein, 0.5%BSA, 0.1%Tween-20,0.05%PEG, 0.1%Tween-80,0.05%PVP, 0.3% 2 hydration lemon
The sour sodium of lemon acid, 2% sucrose.
Abbreviation: BSA: bovine serum albumin(BSA), Casein: casein, Tris-CL: trishydroxymethylaminomethane hydrochloric acid, PEG:
Polyethylene glycol, Tween-20: polysorbas20, Tween-80: Tween 80, PVP: polyvinylpyrrolidone.
Assembling detection card: will coating it is good needed for reagent nitrocellulose filter, offset plate and processed sample pad and
Water absorption pad is pasted together according to mold, and is cut into required width detection card, generally 5mm width, the detection card group sheared
It is attached in corresponding card plug (Cassette);(old system is first nitrocellulose filter and processed sample pad and water suction
Pad is all cut, generally 5mm width, is then sequentially being assembled into corresponding card plug (Cassette)) card plug is in sample pad
There is well in top, and there is observation window in nitrocellulose filter lower section side, acquires for data.With common suction nozzle pipette samples dilution
(sample diluting liquid is more people point mixing, i.e. one big bottle of sample diluting liquid of box product) is to the testing tube for having put fluorescent material
It is interior, then product are loaded, it blows and beats repeatedly for several times, dissolves fluorescent material, and be immunoreacted between measured object in sample.It is then added to card plug
In well, 11 minutes reading data.Or it is first loaded in product to the testing tube for having put fluorescent material, then be loaded product dilution
Liquid is blown and beaten for several times repeatedly, dissolves fluorescent material, and be immunoreacted between measured object in sample.It is then added in card plug well,
Due to capillary action lateral chromatography forward, 11 minutes whens, read data.
When doing human muscle hemoglobin (MYO) project, since nitrocellulose filter shortens, climbs water and obviously become faster, specifically look at
Following table:
Explain: this method is climbed water and is improved 10 seconds.
Embodiment 3
Coated antibody: troponin (cTnI) detection antibody 1 is diluted to 3.0mg/ml concentration with coating buffer, is compareed
Reagent 1 (goat-anti chicken IgY) is diluted to 3.0mg/ml concentration, will be upper using the XYZ3060 of special coating equipment BIODOT company
It states in two liquid coatings to Sai Duolisi nitrocellulose filter (NC), 37 DEG C of drying boxes are 4 hours dry, spare.Coating buffering
Liquid be 0.01Mol/l phosphate buffer (PBs) plus 3% sucrose as protective agent.
Labelled antibody: troponin (cTnI) is detected into antibody 2 and contrast agents 2 (chicken IgY) latex fluorescent marker, storage
There are in storing liquid, spare, (50mM Tris, 0.5%BSA, pH 7.8).
Marker prepares: by above-mentioned labelled antibody by 5 microlitres of volumes to suction nozzle inner wall of required concentration specking, being dried
It is spare.
Sample pad preparation: sample treatment liquid is impregnated by required concentration or specking is in or on sample pad, drying for standby.One
The formula of a optimization are as follows: wherein Tris-CL is molar concentration, remaining is mass percent.Specific formula is as follows: 50mMol/l
Tris-CL, 0.5%Casein, 0.5%BSA, 0.1%Tween-20,0.05%PEG, 0.1%Tween-80,0.05%PVP,
0.3% 2 citric acid monohydrate acid sodium, 2% sucrose.
Abbreviation: BSA: bovine serum albumin(BSA), Casein: casein, Tris-CL: trishydroxymethylaminomethane hydrochloric acid, PEG:
Polyethylene glycol, Tween-20: polysorbas20, Tween-80: Tween 80, PVP: polyvinylpyrrolidone.
Assembling detection card: the nitrocellulose filter and offset plate of reagent needed for new system will be coated with well and processed sample
Pad and water absorption pad are pasted together according to mold, and are cut into required width detection card, generally 5mm width, the inspection sheared
Card is surveyed to be assembled into corresponding card plug (Cassette);Old system first nitrocellulose filter and processed sample pad and
Water absorption pad is all cut, generally 5mm width, is then sequentially being assembled into corresponding card plug (Cassette).Card plug is in sample
There is well above product pad, have observation window above nitrocellulose filter, is acquired for data.With with marker fluorescent material
In suction nozzle pipette samples to reaction buffer bottle, blows and beats repeatedly for several times, dissolve fluorescent material, and between measured object in sample
Immune response.It is then added in card plug well, due to capillary action lateral chromatography forward,
Data are read at 19 minutes.
When doing troponin (cTnI) project, this system precision is significantly improved, and specifically looks at following table:
Explain: either fluorescent value or detectable concentration, this method coefficient of variation (CV) are at least enhanced about more than once, i.e., originally
Method precision improves significantly.
The not most place of the present invention, those skilled in the art, which can according to need, selects existing technological means to complete, than
Such as, the specific location of band and quality control band is detected, specific location of well etc., details are not described herein.
The above description is merely a specific embodiment, but scope of protection of the present invention is not limited thereto, any
Those familiar with the art in the technical scope disclosed by the present invention, can easily think of the change or the replacement, and should all contain
Lid is within protection scope of the present invention.Therefore, protection scope of the present invention should be based on the protection scope of the described claims.
Claims (6)
1. a kind of immune lateral chromatography detection system characterized by comprising
Bottom plate comprising for carrying the first offset plate of sample pad, nitrocellulose filter and the second glue for carrying water absorption pad
Plate;First offset plate and the second offset plate is fixedly connected with the both ends of the nitrocellulose filter respectively;The nitric acid
Detection band and quality control band are provided on cellulose membrane;The baseplate width is 4-6mm, length 6.5-7.5cm;
Sample pad is fixed on first offset plate and contacts with the nitrocellulose filter to guarantee to climb water;
Water absorption pad is fixed on second offset plate and contacts with the nitrocellulose filter to guarantee to climb water;
The immune lateral chromatography detection system further includes card plug comprising:
Kerve is used to fix the bottom plate;
Upper cover is provided with the well for being loaded in the sample pad;
Window is detected, is used to detect the detection window of the detection band and quality control band;The detection window is arranged on kerve.
2. immune lateral chromatography detection system according to claim 1, which is characterized in that the length of first offset plate
For 1.8-2.0cm;The length of second offset plate is 1.5-1.7cm;The length of the sample pad is 1.7-1.8cm;Institute
The length for the water absorption pad stated is 1.2-1.4cm.
3. a kind of preparation method of immune lateral chromatography detection system, which comprises the steps of:
(1) the coating detection band and quality control band on nitrocellulose filter, by the two sides of nitrocellulose filter respectively with offset plate A and glue
Plate B is fixedly connected, and the detection band is between offset plate A and quality control band;
(2) sample pad is fixed on the offset plate A and is contacted with the nitrocellulose filter to guarantee to climb water, will absorbed water
Pad is fixed on the offset plate B and contacts with the nitrocellulose filter to guarantee to climb water;Obtain detection system blank;
(3) the detection system blank is cut into required size up to the immune lateral chromatography detection system;
It further includes following steps that the detection system blank, which is cut into after required size:
It is put into card plug after the detection system blank is cut into required size, the card plug includes:
Kerve is used to fix the bottom plate;The kerve bottom is equipped with for detecting the detection band and quality control band
Detect window;
Upper cover is provided with the well for being loaded in the sample pad.
4. the preparation method of immune lateral chromatography detection system according to claim 3, which is characterized in that the step
(2) specifically comprise the following steps:
With the fixed bottom plate of " work " pattern tool, the sample pad is pasted on offset plate A and pushes down the nitric acid fibre
Plain film is tieed up, pushing down width is 1-3mm;The water absorption pad is pasted on offset plate B to and is pushed down the nitrocellulose filter, pressure
Firmly width is 1-3mm.
5. the preparation method of immune lateral chromatography detection system according to claim 3, which is characterized in that the offset plate
A width is 1.8-2.0cm, and the width of the nitrocellulose filter is 3.3-3.8cm, and the width of the offset plate B is 1.5-
1.7cm。
6. the preparation method of immune lateral chromatography detection system according to claim 3, which is characterized in that described is required
It is 4-6mm having a size of width.
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