CN209400548U - The fluorescence immune chromatography kit of quantitative detection SAA and CRP - Google Patents
The fluorescence immune chromatography kit of quantitative detection SAA and CRP Download PDFInfo
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- CN209400548U CN209400548U CN201822204641.5U CN201822204641U CN209400548U CN 209400548 U CN209400548 U CN 209400548U CN 201822204641 U CN201822204641 U CN 201822204641U CN 209400548 U CN209400548 U CN 209400548U
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Abstract
The utility model is a kind of kit of the fluorescence immune chromatography about quantitative detection serum amyloid A protein and C reactive protein.The kit includes getting stuck；Fluorescence immune chromatography test paper bar, sample pad and nitrocellulose filter including overlap joint setting；The nitrocellulose filter is disposed with the first detection line, the second detection line and nature controlling line from one end of connection sample pad；The fluorescence immune chromatography test paper bar is connected in described get stuck；Buffer unit is connected to described one end got stuck comprising buffer pedestal；The buffer unit further includes the buffer container being connected on the buffer pedestal；The buffer container open end forms sealing structure with aluminium foil sealing.The kit detects CRP and SAA concentration using red nano fluorescent microsphere labelled antibody simultaneously, and luminous intensity is high, emission spectrum is narrow, fluorescence lifetime is long, and testing result high sensitivity, accuracy are strong, being capable of quick diagnosis inflammation infection.
Description
Technical field
The utility model belongs to medical domain, in particular to a kind of quantitative detection serum amyloid A protein (SAA) and C- are anti-
The fluorescence immune chromatography kit of albumen (CRP) is answered, serum amyloid A protein and C- reaction egg can be rapidly and quantitatively detected
It is white.
Background technique
C reactive protein: CRP (C-Reactive protein) is after human body is invaded by a variety of factors such as microorganisms, liver
Dirty to generate a kind of acute phase reactive protein within a few hours, conventional CRP 6-8h after infection occurs starts to increase, and 24-48h reaches
To peak, up to hundreds times of normal value, the hurried decline of its content after infection is eliminated can restore normal peak value in one week.
This is provided an important basis for the identification of disease early infection type.Conventional CRP detection is mainly used for the reality of infectious diseases
Test inspection foundation.
Serum amyloid A protein (ser μm of amyloid A, SAA) is the precursor substance of tissue amyloid A, is belonged to
Acute reaction protein.SAA is in blood with normal component existing for low-level.When human body has virus or bacterial infection scorching
It is increased quickly when disease, activity lesion, tissue mass lesions.Especially in viral acute infection, (48~72h) can be rapid
It increases, and declines rapidly in the convalescence of disease.Therefore SAA is auxiliary diagnostic index very effective when infecting body virus.
Research shows that CRP/SAA concentration level and many severity of disease are in certain positive correlation in blood, it is
The detection project of many disease auxiliary diagnosis, joint-detection CRP/SAA concentration level can more enough effective predictive diseases and its hairs
The degree of danger of disease, the rising of CRP/SAA level implies body there are inflammation infections or disease to be in active stage in body；This
Outer CRP/SAA joint-detection can effectively identify, diagnose bacterium and virus infection, especially infant infection disease morning
Phase, bacterium and virus are difficult to discriminate between, and the variation of joint-detection serum CA125 and SAA level and SAA/CRP ratio facilitates
The antidiastole of children's bacterium infection and virus infection.Detection sensitivity and specificity can be improved in CRP and SAA joint-detection, right
Antidiastole provides more fully foundation, and the two joint-detection, which has, to have complementary advantages and clinical value-added meaning, than individually detecting
One project is more meaningful.
However, not occurring the product of CRP/SAA simultaneous quantitative joint-detection currently on the market, clinically to patient's
Detection can only be detected item by item while CRP/SAA concentration.On the one hand it takes time and effort, on the other hand causes reagent, consumptive material
Waste, increase detection treatment cost.
Utility model content
The main purpose of the utility model is to provide a kind of quantitative detection serum amyloid A proteins (SAA) and C- to react
When body is in Infection Status, amount exists as unit of mg/L by the fluorescence immune chromatography kit of albumen (CRP), CRP and SAA
Exist in body, the condition for having while detecting；The kit uses fluorescence immunoassay lateral chromatography technology, rapidly and quantitatively
Detect serum amyloid A protein and C reactive protein；The detection kit uses red nano fluorescent microsphere labelled antibody,
Its luminous intensity is high, emission spectrum is narrow, fluorescence lifetime is long, and testing result high sensitivity, accuracy be strong, being capable of quick diagnosis inflammation
Infection is suitable for situation of all-level hospitals and domestic medicine, thus more suitable for practical.
It the purpose of this utility model and solves its technical problem and adopts the following technical solutions to realize.It is practical according to this
The fluorescence immune chromatography kit of a kind of the quantitative detection serum amyloid A protein and C reactive protein of novel proposition comprising
It gets stuck；Fluorescence immune chromatography test paper bar, sample pad and nitrocellulose filter including overlap joint setting；The nitrocellulose
Film is disposed with the first detection line, the second detection line and nature controlling line from one end of connection sample pad；The fluorescence immunoassay layer
Analysis test strips are connected in described get stuck；Buffer unit is connected to described one end got stuck comprising buffer pedestal；Institute
Stating buffer unit further includes the buffer container being connected on the buffer pedestal；It uses the open end of the buffer container
Aluminium foil sealing forms sealing structure；It is packaged with serum amyloid A protein in the buffer container and C reactive protein sample is slow
Fliud flushing；The first anti-human specificity of serum amyloid A protein mouse of red nano fluorescent microsphere label is fixed in the sample pad
The anti-human specific antibody of the first C reactive protein mouse and red nano fluorescent microsphere mark of antibody, red nano fluorescent microsphere label
The chicken IgY antibody of note；The second C reactive protein that red nano fluorescent microsphere label is fixed in first detection line is special
Property antibody specificity antibody；The second serum amyloid sample egg of red nano fluorescent microsphere label is fixed in second detection line
White A specific antibody；Chicken endogenous antibody is fixed on the nature controlling line.
It the purpose of this utility model and solves its technical problem also following technical measures can be used to further realize.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the fluorescence immune chromatography test paper bar is provided with the bottom plate comprising nearly sample end and remote sample end, on the bottom plate by
Nearly sample end is successively overlapped to remote sample end is provided with sample pad, nitrocellulose filter and water absorption pad.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the Ka Gai to get stuck including card bottom and lid on it；The fluorescence immune chromatography test paper bar is placed in the card bottom
It is interior；Being internally provided with for the Ka Gai is several for compressing the stuck point of the fluorescence immune chromatography test paper bar；The card covers also
It is provided with well and observation window；The sample pad is located at the lower section of the well；First detection line, the second detection line
It is located at the lower section of the observation window with nature controlling line.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the excitation-emission wavelength of the red nano fluorescent microsphere is to for 570-590nm and 610-620nm.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the second serum amyloid A protein specific antibody is mouse anti-human serum amyloid A monoclonal antibody；Institute
Stating the second C reactive protein specific antibody is the anti-human C reactive protein monoclonal antibody of mouse.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the chicken endogenous antibody is goat-anti chicken IgY polyclonal antibody.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the serum amyloid A protein and C reactive protein sample buffer in the buffer container include several person-portions
Buffer；Single part volume of the buffer is
100-1500uL。
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box wherein the bottom plate is white, and is attached with adhesive sticker；It is described get stuck, nitrocellulose filter, bottom plate and adhesive sticker are free of
There is fluorescer.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the volume of the sample to be tested of the kit is 5-40uL.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the bottom plate uses PVC offset plate, having a size of length 6-8cm, width 4-6mm, thickness 0.25-0.5mm.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the sample pad uses glass fibre filter membrane, having a size of length 15-30mm, width 4-6mm, thickness 0.5-
1mm。
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the size of the nitrocellulose filter is length 20-35mm, width 4-6mm, thickness 0.2-0.3mm.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the size of the water absorption pad is length 10-30mm, width 4-6mm, thickness 0.9-1.8mm.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the distance between first detection line and well orthographic projection position are 15-25mm.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein second detection line is apart from the first detection line 3-6mm.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the nature controlling line is apart from the second detection line 3-6mm.
By above-mentioned technical proposal, the utility model proposes a kind of quantitative detection serum amyloid A protein and C- reaction
The fluorescence immune chromatography kit of albumen at least has the advantage that
1) kit of the utility model uses red nano fluorescent microsphere labelled antibody, not only can be to avoid blood background
The interference of fluorescent material, and have that luminous intensity is high, emission spectrum is narrow, fluorescence lifetime is long, surface modification multifunction, stabilization
Property the advantages such as good and sensitivity height；
2) getting stuck in the kit forms component of the utility model, bottom plate and nitrocellulose filter are without fluorescence
Matter, it is possible to reduce to the influence that fluorescent microsphere fluorescence signal obtains, to guarantee to obtain high fluorescence signal-to-background ratio, and then reach and mention
Highly sensitive purpose；
3) kit of the utility model using fluorescence immune chromatography method rapidly and quantitatively detect serum amyloid A protein and
C reactive protein (SAA/CRP) can reduce non-specific adsorption, reduce autofluorescent background, enhancing specific binding, and then improve inspection
Survey sensitivity, when being conducive to serum amyloid A protein and extremely low C reactive protein (SAA/CRP) content in sample it is accurate calmly
Amount；
4) kit of the utility model is compared to conventional fluorescent immune chromatography reagent kit, have label stability it is good,
Non-specific low, high sensitivity, the range of linearity are wide and quantify the advantages such as accurate；
5) kit of the utility model being capable of CRP and SAA concentration in accurate detection blood, and two kinds of antigens simultaneously
It does not interfere with each other, is as a result read by instrument, have many advantages, such as quick, simplicity, operated without professional；On the one hand the time is saved
And manpower, detection reagent and consumptive material are on the other hand saved, the cost of detection treatment is significantly reduced；
6) detection sensitivity and specificity can be improved for the joint-detection of CRP/SAA in the kit of the utility model, right
Antidiastole provides more fully foundation, and the two joint-detection, which has, to have complementary advantages and clinical value-added meaning, than individually detecting
One project is more meaningful.
The above description is merely an outline of the technical solution of the present invention, in order to better understand the skill of the utility model
Art means, and can be implemented in accordance with the contents of the specification, below on the preferred embodiment of the present invention and the accompanying drawings in detail
It describes in detail bright as after.
Detailed description of the invention
Fig. 1 is the kit overlooking structure diagram of the utility model；
Fig. 2 is the A-A the schematic diagram of the section structure of Fig. 1；
Fig. 3 is the structural schematic diagram of the fluorescence immune chromatography test paper bar of the utility model.
Specific embodiment
Further to illustrate that the utility model is the technical means and efficacy reaching predetermined goal of the invention and being taken, below
In conjunction with attached drawing and preferred embodiment, to according to the utility model proposes a kind of quantitative detection serum amyloid A protein and C- it is anti-
The fluorescence immune chromatography kit of albumen is answered, specific embodiment, structure, feature and its effect, detailed description is as follows.
As shown in attached drawing 1 to attached drawing 3, the utility model proposes a kind of quantitative detection serum amyloid A protein and C- it is anti-
Answer the fluorescence immune chromatography kit of albumen comprising get stuck 1；Fluorescence immune chromatography test paper bar 2, the sample including overlap joint setting
Product pad 22 and nitrocellulose filter 23；The nitrocellulose filter 23 is disposed with from one end of connection sample pad 22
First detection line 231, the second detection line 232 and nature controlling line 233；The fluorescence immune chromatography test paper bar 2, which is connected in, described gets stuck 1
It is interior；Buffer unit 3, be connected to it is described get stuck 1 one end comprising buffer pedestal；The buffer unit 3 further includes card
The buffer container being connected on the buffer pedestal；The open end of buffer container aluminium foil sealing forms sealing knot
Structure；Serum amyloid A protein and C reactive protein sample buffer are packaged in the buffer container；In the sample pad 22
It is fixed with the anti-human specific antibody of the first serum amyloid A protein mouse, the red nano fluorescence of red nano fluorescent microsphere label
The anti-human specific antibody of the first C reactive protein mouse of microballoon label and the chicken IgY antibody of red nano fluorescent microsphere label；Institute
State be fixed in the first detection line 231 red nano fluorescent microsphere label the second C reactive protein specific antibody specificity it is anti-
Body；The second serum amyloid A protein specificity that red nano fluorescent microsphere label is fixed in second detection line 232 is anti-
Body；Chicken endogenous antibody is fixed on the nature controlling line 233.
It is described get stuck 1 be the fluorescence immune chromatography test paper bar 2 external shell structure.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the fluorescence immune chromatography test paper bar 2 is provided with the bottom plate 21 including nearly sample end and remote sample end, the bottom plate
It is successively overlapped on 21 by nearly sample end to remote sample end and is provided with sample pad 22, nitrocellulose filter 23 and water absorption pad 24.
When the fluorescence immune chromatography kit of the quantitative detection serum amyloid A protein and C reactive protein is for examining
When surveying serum amyloid A protein and C reactive protein, sample to be tested is serum, blood plasma or whole blood.The sample to be tested is through institute
The well 111 stated is added in sample pad 22, the first blood marked in the sample pad with red nano fluorescent microsphere
Clear amyloid A and C reactive protein specific antibody combine, and chromatograph and transport to 24 direction of water absorption pad along nitrocellulose filter 23
It is dynamic, the chromatography time preferably 5 minutes.The sample pad 22 of the fluorescence immune chromatography test paper bar 2 has sample simultaneously at this time
The effect of product collection, combination, release and filtering.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the 1 card lid 11 including card bottom 12 and lid on it that gets stuck；The fluorescence immune chromatography test paper bar 2 is placed in institute
It states in card bottom 12；Being internally provided with for the card lid 11 is several for compressing the stuck point of the fluorescence immune chromatography test paper bar 2；Institute
It states and is additionally provided with well 111 and observation window 112 on card lid 11；The sample pad 22 is located at the lower section of the well 111；Institute
State the lower section that the first detection line 231, the second detection line 232 and nature controlling line 233 are located at the observation window 112.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the excitation wavelength of the red nano fluorescent microsphere is 570-590nm, launch wavelength 610-620nm.
The red nano fluorescent microsphere can be the fluorescent microsphere of single compound or be made of several compounds
Composite fluorescence microballoon；The preferred single compound fluorescent microsphere of the fluorescent microsphere and preferably have stronger photostability
Fluorescent microsphere.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the second serum amyloid A protein specific antibody is mouse anti-human serum amyloid A monoclonal antibody；Institute
Stating the second C reactive protein specific antibody is the anti-human C reactive protein monoclonal antibody of mouse.
The second serum amyloid A protein specific antibody is coated in second detection line 232.
The second C reactive protein specific antibody is coated in first detection line 231.
First serum amyloid A protein of fixed red nano fluorescent microsphere label and C- reaction in the sample pad
The of coated red nano fluorescent microsphere label in albumen (SAA/CRP) specific antibody and second detection line 232
The 2nd C- of coated red nano fluorescent microsphere label is anti-on two serum amyloid A proteins and first detection line 231
The antigenic determinant for answering protein specific antibody to be identified is different.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the chicken endogenous antibody is the anti-chicken IgY polyclonal antibody of sample.
The chicken endogenous antibody is coated on the nature controlling line 233, to capture extra fluorescent microsphere.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein serum amyloid A protein and C reactive protein sample buffer in the buffer container include several person-portions
Buffer；Single part volume of the buffer is 100-1500uL.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the bottom plate 21 is white, and surface has adhesive sticker；The sample pad 22, nitrocellulose filter 23 and water suction
Pad 24 is bonded on the bottom plate 21 by adhesive sticker；The 1, nitrocellulose filter 23 that gets stuck, bottom plate 21 and adhesive sticker are equal
Without containing fluorescer.
In order to reduce influence of the kit to fluorescent microsphere fluorescence signal, the utility model uses the nitric acid without fluorescence fine
Tie up plain film 23, bottom plate 21 and get stuck 1, to guarantee to obtain high fluorescence signal-to-background ratio, can good discrimination signal and background, Jin Erti
High detection sensitivity.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the volume of the sample to be tested of the kit is 5-40uL.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the bottom plate 21 uses PVC offset plate, having a size of length 6-8cm, width 4-6mm, thickness 0.25-0.5mm.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the sample pad 22 uses glass fibre filter membrane, having a size of length 15-30mm, width 4-6mm, thickness
0.5-1mm。
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the size of the nitrocellulose filter 23 is length 20-35mm, width 4-6mm, thickness 0.2-0.3mm.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the size of the water absorption pad 24 is length 10-30mm, width 4-6mm, thickness 0.9-1.8mm.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the distance between 111 orthographic projection position of first detection line 231 and well is 15-25mm.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein second detection line 232 is 3-6mm at a distance from first detection line 231.
Preferably, the reagent of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein above-mentioned
Box, wherein the nature controlling line 233 is 3-6mm at a distance from second detection line 232.
The water absorption pad 24 uses the mixing material of glass and velveteen.
The preparation side of the fluorescence immune chromatography kit of above-mentioned quantitative detection serum amyloid A protein and C reactive protein
Method is as follows:
1) fluorescent labeled antibody:
Red nano fluorescent microsphere is anti-with serum amyloid A protein (SAA), C reactive protein (CRP) monoclonal respectively
Body and the mixing of chicken IgY antibody, ultrasonic reaction 2h are added 1mol/L glycine and terminate reaction, and cleaning saves, and it is glimmering to obtain red nano
C reactive protein (CRP) monoclonal that serum amyloid A protein (SAA), the red nano fluorescent microsphere of light microballoon label mark
The chicken IgY antibody of antibody and red nano fluorescent microsphere label.Used chemical cross-linking agent includes 1- ethyl -3- (3- diformazan
Base amine propyl) carbodiimides (EDC) or n-hydroxysuccinimide (NHS) etc..
After anti-Miao Le Shi pipe hormone (AMH) specific antibody label of the red nano fluorescent microsphere label, first
It is stored in storing liquid, it is spare；The storing liquid is 50 mMs of Tris and 0.5%BSA, pH value 7.8.
The serum amyloid A protein and C reactive protein of the label of red nano fluorescent microsphere used by the utility model
(SAA/CRP) specific antibody is the product of Beijing O&D Biotech Co. Ltd..
2) building of fluorescence immune chromatography test paper bar:
A, by sample pad using sample treatment liquid by the immersion of required concentration, or sample treatment liquid is sprayed in sample pad or
Sample pad surface, drying for standby.Preferred sample treatment liquid includes: 50 mMs of Tris-HCl, 0.5wt%Casein,
Bis- water of 0.5wt%BSA, 0.1wt%Tween-20,0.05wt%PEG, 0.1wt%Tween-80,0.05%PVP, 0.3wt%
Close citric acid sodium and 2wt% sucrose, wherein Tris-HCl is " trishydroxymethylaminomethane hydrochloride buffer ", and Casein is
" casein ", BSA are " bovine serum albumin(BSA) ", and PEG is " polyethylene glycol ", and Tween-20 is " polysorbas20 ", and Tween-80 is " to spit
80 ", PVP of temperature is " polyvinylpyrrolidone ".
B, the first SAA antibody, the first CRP antibody and the chicken IgY antibody for marking red nano fluorescent microsphere, it is micro- with fluorescence
Ball dilution sprays in sample pad in proportion, and 37 DEG C of dry 2h, 2-30 DEG C saves backup；
First SAA antibody marker, the first CRP antibody marker and the chicken IgY marker is mixed in the ratio of 2:2:1
It closes in marker dilution, above-mentioned handle well is sprayed on using special spray marker equipment (XYZ3060 of BIODOT company)
Sample pad on, drying for standby；Make final every person-portion containing the first SAA antibody marker 1-5 μ g, the first CRP antibody marker 1-5
μ g, chicken IgY marker 0.5-2.5 μ g.The marker dilution includes: 50 mMs of Tris-HCl, 2wt%BSA,
0.5wt%PEG, 0.1wt%PVP, 6wt% mannitol and 8wt% trehalose.
C, first detection line, second detection line and a nature controlling line are respectively set on nitrocellulose filter,
Wherein, red can be fixed on the chicken endogenous antibody in conjunction with Quality Control molecular specificity, the first detection line by being fixed on nature controlling line
The second C reactive protein (CRP) specific antibody of nanometer fluorescent microspheres label, it is glimmering to be fixed with red nano in the second detection line
The second serum amyloid A protein (SAA) specific antibody of light microballoon label；And second C reactive protein (CRP) specificity it is anti-
The antigenic determinant and the red being fixed in sample pad that body, the second serum amyloid A protein (SAA) specific antibody are identified
The anti-human specific antibody of the first serum amyloid A protein (SAA) mouse and the first C reactive protein of nanometer fluorescent microspheres label
(CRP) antigenic determinant that the anti-human specific antibody of mouse is identified is different；
In a preferred scheme, by the second C reactive protein (CRP) specific antibody, the second serum amyloid protein
A (SAA) specific antibody and Quality Control antibody are diluted to the fixed concentration of 0.5-3.0mg/ml with coating buffer respectively；Described
It is coated with the phosphate buffer (PBS) that buffer is 0.01M, adds 3% sucrose as protective agent；The coating is using special
Above three liquid is coated on nitrocellulose filter (detection film) by the XYZ3060 for being coated with equipment-BIODOT company, 37 DEG C
It is 4 hours dry in drying box, it is spare.
D, sample pad, nitrocellulose filter, water absorption pad are successively equipped on bottom plate and are built into fluorescence immune chromatography
Test strips；
3) it is loaded:
Fluorescence immune chromatography test paper bar loading is got stuck interior, installation direction needs so that described in the well face that card covers
Sample pad, the first detection line, the second detection line and nature controlling line described in observation window face；
4) buffer unit encapsulates:
The serum amyloid A protein and C reactive protein template buffering of designer part are packed into the buffer container
Liquid, aluminium foil sealing form closed structure；Then the closed structure is connected on the buffer pedestal；
The sample buffer includes: that 50 mMs of Tris-HCl, 0.1wt%Tween-80 and 0.73wt% bis- are hydrated
Trisodium citrate.
The detection of the utility model uses fluorescent quantitation instrument.The fluorescent quantitation instrument includes excitation light source module, filters
Module, photoelectric conversion module, control analysis module and software systems.Wherein excitation light source module includes that light source and optically focused fill
It sets, which is light emitting diode or laser diode, and wavelength is between 550~750nm.Filtration module includes optical filter
Wheel, which includes optical filter not of the same race, to obtain corresponding fluorescence signal.Photoelectric conversion module includes imaging sensor
Or photomultiplier tube.
Sample to be tested is added dropwise in sample pad by well, the sample to be tested and red nano fluorescent microsphere mark
Note the first serum amyloid A protein, red nano fluorescent microsphere label C reactive protein (SAA/CRP) specific antibody and
The chicken IgY antibody of red nano fluorescent microsphere label is chromatographed after being combined in sample pad；After chromatographing, use
The light source that wavelength is 570-590nm irradiates the test strips, and the fluorescent material in the test strips launches wavelength and is
The fluorescence of 610-620nm.The fluorescence signal that the test strips generate filters out stray light and background fluorescence through filtration module, reaches
Photoelectric conversion module obtains digital signal.The fluorescence signal intensity that the fluorescence signal intensity and detection line that nature controlling line obtains obtain
With correlation: if nature controlling line fluorescence signal intensity exceeds the acceptable value of fluorescent quantitation instrument inner setting, illustrating detection knot
Fruit is invalid；Under the premise of testing result is effective, the ratio of fluorescence signal intensity and nature controlling line fluorescence signal intensity that detection line obtains
Value is higher, indicates that the concentration of target detection thing in sample is higher, otherwise lower.
Blood of human body itself has a fluorescent characteristic, excitation-emission to mainly have 260-630nm, 280-340nm,
340-460nm, 450-520nm, corresponding to Endogenous fluorophores be respectively porphyrin, tryptophan, reduction nicotinamide adenine
Therefore how dinucleotides and yellow adenine-dinucleotide in using detection of the blood as sample to be tested, avoid blood
Middle Endogenous fluorophores are the problem of having to think deeply to the interference of testing result.
The kit of the utility model uses red nano fluorescent microsphere labelled antibody, uses wavelength for 570-590nm's
Light source irradiates the test strips, and the fluorescent material in the test strips launches the fluorescence that wavelength is 610-620nm, not only
Can be to avoid the interference of blood background fluorescence substance, and it is high with luminous intensity, emission spectrum is narrow, fluorescence lifetime is long, table
Face modifies that multifunction, stability is good and the advantages such as sensitivity height.
The utility model detects a series of standard items of various concentrations using fluorescent quantitation instrument, draws standard curve.Then
Test sample, establishing criteria curve obtain the dense of serum amyloid A protein and C reactive protein (SAA/CRP) in sample to be tested
Degree.
The fluorescence immune chromatography kit of above-mentioned quantitative detection serum amyloid A protein and C reactive protein is for detecting
When, it follows the steps below:
A) normal human serum is used to prepare gradient calibration object respectively as dilution SAA/CRP antigen standard；
B) buffer container is added in the 5 μ L of serum solution prepared in step a), making it, sufficiently piping and druming is mixed in buffer
It is even, react 5min；
C) it places it in and obtains fluorescence signal intensity in fluorescent quantitation instrument, and draw corresponding standard curve；
D) corresponding lot card is written into standard curve；
E) it is operated with step b), sample to be tested is detected, after chromatography, be placed in fluorescent quantitation instrument and obtain fluorescence
Signal strength, instrument analyze the content of SAA/CRP in the sample automatically according to the standard curve of lot card；
It is detected according to above-mentioned detecting step, the results showed that, C reactive protein lowest detection line is 0.5mg/L, blood
Clear amyloid A lowest detection is limited to 5mg/L, and batch in batch between preferable, coefficient R > 0.99 of repeatability.By above
The fluorescence immune chromatography for showing quantitative detection serum amyloid A protein and C reactive protein that the utility model is proposed
Kit provides reference to Evaluation of Inflammation.
The working principle of the fluorescence immune chromatography kit of the utility model are as follows: using fluorescence immune chromatography technology and dual anti-
Serum amyloid A protein and C reactive protein (SAA/ in body sandwich method principle quantitative detection sample (whole blood, serum or blood plasma)
CRP content).When detection, sample is added drop-wise in well, is mutually tied in sample pad with red nano Fluorescent microsphere marker
It closes, along nitrocellulose filter to water absorption pad direction capillary moving, flows separately through the first detection line on nitrocellulose filter, the
Two detection lines and nature controlling line.It is and red if containing serum amyloid A protein and C reactive protein (SAA/CRP) in sample
The serum amyloid A protein and C reactive protein (SAA/CRP) antibody of nanometer fluorescent microspheres label combine, when chromatography to detection
When line, the capture antibody that can be coated in this detection line in advance is captured, to constitute double-antibody sandwich compound.Using spy
Under the long light source activation of standing wave, the fluorescence signal intensity of detection line and nature controlling line can be obtained using fluorescent quantitation instrument, according to fluorescence
The standard curve that quantitative instrument obtains, and then the concentration in sample containing SAA/CRP can be analyzed.
The above descriptions are merely preferred embodiments of the present invention, not makees in any form to the utility model
Limitation, any simple modification, equivalent change and modification made by the above technical examples according to the technical essence of the present invention,
It is still within the scope of the technical solutions of the present invention.
Claims (10)
1. a kind of kit of the fluorescence immune chromatography of quantitative detection serum amyloid A protein and C reactive protein, feature exist
In comprising:
It gets stuck；
Fluorescence immune chromatography test paper bar, sample pad and nitrocellulose filter including overlap joint setting；The nitrocellulose
Film is disposed with the first detection line, the second detection line and nature controlling line from one end of connection sample pad；The fluorescence immunoassay layer
Analysis test strips are connected in described get stuck；
Buffer unit is connected to described one end got stuck comprising buffer pedestal；The buffer unit further includes clamping
Buffer container on the buffer pedestal；The open end of the buffer container forms sealing structure with aluminium foil sealing；
Serum amyloid A protein and C reactive protein sample buffer are packaged in the buffer container；
The anti-human specificity of the first serum amyloid A protein mouse that red nano fluorescent microsphere label is fixed in the sample pad is anti-
Body, the anti-human specific antibody of the first C reactive protein mouse of red nano fluorescent microsphere label and red nano fluorescent microsphere label
Chicken IgY antibody；
The second C reactive protein specific antibody that red nano fluorescent microsphere label is fixed in first detection line is special
Property antibody；
The second serum amyloid A protein specificity that red nano fluorescent microsphere label is fixed in second detection line is anti-
Body；
Chicken endogenous antibody is fixed on the nature controlling line.
2. the fluorescence immune chromatography of quantitative detection serum amyloid A protein according to claim 1 and C reactive protein
Kit, which is characterized in that
The fluorescence immune chromatography test paper bar is provided with the bottom plate comprising nearly sample end and remote sample end, by nearly sample on the bottom plate
Product end successively overlaps to remote sample end and is provided with sample pad, nitrocellulose filter and water absorption pad.
3. the fluorescence immune chromatography of quantitative detection serum amyloid A protein according to claim 1 and C reactive protein
Kit, which is characterized in that
The Ka Gai to get stuck including card bottom and lid on it；The fluorescence immune chromatography test paper bar is placed in the card bottom；
Being internally provided with for the Ka Gai is several for compressing the stuck point of the fluorescence immune chromatography test paper bar；The card, which covers, to be also set up
There are well and observation window；The sample pad is located at the lower section of the well；First detection line, the second detection line and matter
Control line is located at the lower section of the observation window.
4. the fluorescence immune chromatography of quantitative detection serum amyloid A protein according to claim 1 and C reactive protein
Kit, which is characterized in that
The excitation-emission wavelength of the red nano fluorescent microsphere is to for 570-590nm and 610-620nm.
5. the fluorescence immune chromatography of quantitative detection serum amyloid A protein according to claim 1 and C reactive protein
Kit, which is characterized in that
The second serum amyloid A protein specific antibody is mouse anti-human serum amyloid A monoclonal antibody；
The second C reactive protein specific antibody is the anti-human C reactive protein monoclonal antibody of mouse.
6. the fluorescence immune chromatography of quantitative detection serum amyloid A protein according to claim 1 and C reactive protein tries
Agent box, which is characterized in that
The chicken endogenous antibody is goat-anti chicken IgY polyclonal antibody.
7. the fluorescence immune chromatography of quantitative detection serum amyloid A protein according to claim 1 and C reactive protein tries
Agent box, which is characterized in that
Serum amyloid A protein and C reactive protein sample buffer in the buffer container include the buffering of several person-portions
Liquid；Single part volume of the buffer is 100-1500uL.
8. the fluorescence immune chromatography of quantitative detection serum amyloid A protein according to claim 2 and C reactive protein tries
Agent box, which is characterized in that
The bottom plate is white, and is attached with adhesive sticker；
It is described get stuck, nitrocellulose filter, bottom plate and adhesive sticker do not contain fluorescer.
9. the fluorescence immune chromatography of quantitative detection serum amyloid A protein according to claim 1 and C reactive protein tries
Agent box, which is characterized in that
The volume of the sample to be tested of the kit is 5-40uL.
10. the fluorescence immune chromatography of quantitative detection serum amyloid A protein according to claim 2 and C reactive protein tries
Agent box, which is characterized in that
The bottom plate uses PVC offset plate, having a size of length 6-8cm, width 4-6mm, thickness 0.25-0.5mm；And/or
The sample pad uses glass fibre filter membrane, having a size of length 15-30mm, width 4-6mm, thickness 0.5-1mm；
And/or
The size of the nitrocellulose filter is length 20-35mm, width 4-6mm, thickness 0.2-0.3mm；And/or
The size of the water absorption pad is length 10-30mm, width 4-6mm, thickness 0.9-1.8mm；And/or
The distance between first detection line and well orthographic projection position are 15-25mm；And/or
Second detection line is apart from the first detection line 3-6mm；And/or
The nature controlling line is apart from the second detection line 3-6mm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201822204641.5U CN209400548U (en) | 2018-12-26 | 2018-12-26 | The fluorescence immune chromatography kit of quantitative detection SAA and CRP |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201822204641.5U CN209400548U (en) | 2018-12-26 | 2018-12-26 | The fluorescence immune chromatography kit of quantitative detection SAA and CRP |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN209400548U true CN209400548U (en) | 2019-09-17 |
Family
ID=67896387
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201822204641.5U Active CN209400548U (en) | 2018-12-26 | 2018-12-26 | The fluorescence immune chromatography kit of quantitative detection SAA and CRP |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN209400548U (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112485446A (en) * | 2020-11-18 | 2021-03-12 | 重庆中元汇吉生物技术有限公司 | Kit for measuring full-range C-reactive protein and preparation method thereof |
| CN115047187A (en) * | 2022-07-26 | 2022-09-13 | 石家庄希宝生物科技有限公司 | Test strip sample pad treatment solution, preparation method and application thereof |
-
2018
- 2018-12-26 CN CN201822204641.5U patent/CN209400548U/en active Active
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112485446A (en) * | 2020-11-18 | 2021-03-12 | 重庆中元汇吉生物技术有限公司 | Kit for measuring full-range C-reactive protein and preparation method thereof |
| CN115047187A (en) * | 2022-07-26 | 2022-09-13 | 石家庄希宝生物科技有限公司 | Test strip sample pad treatment solution, preparation method and application thereof |
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