CN107656063A - Tilapia mossambica Streptococcusagalactiae immune chromatography rapid detecting test paper strip and preparation method thereof - Google Patents
Tilapia mossambica Streptococcusagalactiae immune chromatography rapid detecting test paper strip and preparation method thereof Download PDFInfo
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
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Abstract
The invention discloses Tilapia mossambica Streptococcusagalactiae immune chromatography rapid detecting test paper strip and preparation method thereof, test strip is made up of supporting plate, sample pad, label pad, coated film, adsorptive pads.The label pad is coated with anti-Tilapia mossambica Streptococcusagalactiae monoclonal antibody 4C12, the coated film upper forepart is provided with detection line, rear portion is provided with nature controlling line on the coated film, the detection line is coated with the monoclonal antibody 3A9 of anti-Tilapia mossambica Streptococcusagalactiae, and the nature controlling line is coated with goat anti-mouse immunoglobulin antibody.The upper sample pad, label pad, coated film, adsorptive pads are overlapped into stickup on the supporting plate successively.Advantage of the present invention：The test strips of the present invention are easy to operate, one of skill in the art and professional equipment instrument are not needed, the needs of the needs of different levels personnel and department, especially basic unit's production unit can be met, and had the advantages that quick, accurate, sensitive, special, cheap.
Description
Technical field
The invention belongs to biological monitoring field, specifically Tilapia mossambica Streptococcusagalactiae immune chromatography rapid detecting test paper strip and
Its preparation method.
Background technology
Tilapia mossambica (Tilapia) is a kind of cultured fishes for having extensive adaptability, and the major protein that the United Nations is recommended
The cultured fishes in matter source.Because it has growth fast, the characteristics of wide is adapted to, is occupied an important position in fish culture.I
State is Rofe fish culture big country, accounts for the 90% of world export amount.Southern region of China is the main producing region of cultivation, accounts for national yield
90%.In recent years, with the industrialized development of Tilapia mossambica aquaculture, disease is also following, particularly Tilapia mossambica agalasisa hammer
Breaking out for bacterium disease, serious loss is brought to aquaculture.Also bring serious influence to outlet simultaneously.
Quickly diagnosis is to prevent and treat the key of Tilapia mossambica Streptococcusagalactiae disease, predominantly detects diagnosis to the disease at present
Method has：Traditional bacteriological method, using PCR as molecular biological testing of representative etc., it is special that these methods are required for
Detecting instrument, while need technical professional to operate, complex operation, while time-consuming, it is impossible to adapt to the inspection in production
Surveying diagnosis needs.In order to fast and effectively control Streptococcusagalactiae sick, reduce the disease to the harm that Tilapia mossambica aquaculture is brought with
Lose, be badly in need of fast and effective easy Tilapia mossambica streptococcosis checkout and diagnosis method in production.The present invention utilizes immune colloid gold
Labelling technique, immunochromatography technique and monoclonal antibody technique, invent a kind of new Streptococcusagalactiae fast diagnosis method, i.e. sieve
The immune chromatography rapid detecting test paper strip method of non-fish Streptococcusagalactiae, this method have simple to operate, quick (5-10 minutes
Inside go out result), it is not necessary to special detecting instrument equipment, it is not necessary to the testing staff of specialized training, and can carry out live fast
Speed detection, the method are especially suitable for basic unit and the detection of clinical Streptococcusagalactiae and the quarantine and monitoring of Streptococcusagalactiae disease.
The content of the invention
The defects of it is an object of the invention to overcome prior art and meet above-mentioned field for Tilapia mossambica Streptococcusagalactiae
Sick field quick detection, diagnosis demand, there is provided a kind of Tilapia mossambica Streptococcusagalactiae disease immune chromatography rapid detecting test paper strip and
Its preparation method.
The technical scheme that the present invention solves above-mentioned technical problem is as follows：
A kind of Tilapia mossambica Streptococcusagalactiae immune chromatography rapid detecting test paper strip, it have selected anti-sieve that we prepare, screened
Non- fish Streptococcusagalactiae antigen monoclonal antibody 4C12 is by preserving number as the coated antibody of label pad, the monoclonal antibody：
CCTCC NO:C2014248, Classification And Nomenclature are thin for a kind of hybridoma for the monoclonal antibody for producing anti-Tilapia mossambica Streptococcusagalactiae
Born of the same parents' strain 4C12, on January 7th, 2015 are stored in China typical culture collection center, address：Wuhan University of Wuhan, China city.Choosing
It is detection line (detection line (T lines)) coated antibody to select anti-Tilapia mossambica Streptococcusagalactiae antigen monoclonal antibody 3A9, and the monoclonal resists
Body is by preserving number:CCTCC NO:C2014244 Classification And Nomenclatures resist for a kind of monoclonal for producing anti-Tilapia mossambica Streptococcusagalactiae
The hybridoma cell strain 3A9 of body, is stored in China typical culture collection center, address on January 7th, 2015：Wuhan, China city
Wuhan University.
Tilapia mossambica Streptococcusagalactiae immune chromatography rapid detecting test paper strip and preparation method thereof, the test strip is by propping up
Fagging 1, sample pad 2, label pad 3, coated film 4, adsorptive pads 5 form.
Anti- Tilapia mossambica Streptococcusagalactiae monoclonal antibody 4C12 is coated with to the label pad 3.By anti-Tilapia mossambica agalasisa
Streptococcic monoclonal antibody 3A9 is coated with to the detection line T lines of the coated film 4.By goat anti-mouse immunoglobulin antibody
It is coated with to the nature controlling line C of the coated film 4.
The sample pad 2, label pad 3, coated film 4, adsorptive pads 2 are overlapped successively and are pasted onto in supporting plate 1, Tilapia mossambica
Streptococcusagalactiae immune chromatography rapid detecting test paper strip is prepared and completed.
The material of above-mentioned sample pad 2 and label pad 3 is glass fibre membrane, and the material of above-mentioned coated film 4 is nitrocellulose
Film, the material of above-mentioned adsorptive pads 5 is crude fibre blotting paper, and the test strips supporting plate 1 is the polyvinyl chloride panel not absorbed water.
The anti-Tilapia mossambica Streptococcusagalactiae monoclonal antibody 4C12 monoclonal antibodies of above-mentioned colloid gold label, which are used to be coated with, to be marked
The amount of note pad is often to pad 5 μ L-30 μ L.
Coating concentration of the 3A9 monoclonal antibodies of above-mentioned anti-Tilapia mossambica Streptococcusagalactiae antigen in detection line (T lines) is
0.6mg/mL, package amount are often pad 1-5 μ L/cm2；Coating of the goat anti-mouse immunoglobulin antibody on nature controlling line (C)
Concentration is 2.0mg/mL, and package amount is often pad 1-5 μ L/cm2。
The preparation method of the Tilapia mossambica Streptococcusagalactiae immune chromatography rapid detecting test paper strip of specific detail, including following step
Suddenly：
1. with above two hybridoma, (preserving number is according to a conventional method：CCTCC NO:C2014248：Preserving number is:
CCTCC NO:C2014244 mouse ascites monoclonal antibody 4C12 and 3A9) are prepared respectively.The anti-Tilapia mossambica Streptococcusagalactiae
The monoclonal antibody 4C12 and 3A9 of antigen are essentially protein, can specific recognition Tilapia mossambica Streptococcusagalactiae.With it is conventional just
Octanoic acid-saturated ammonium sulfate method purification mouse ascites monoclonal antibody 4C12 and 3A9, is detected with BCA determination of protein concentration kit
The content of monoclonal antibody.
2. the collaurum of 20-40nm size particles is prepared using sodium citrate according to a conventional method.Specific method is as follows：
0.01% secondary gold chloride is added in 250ml conical flask, is heated on electromagnetic constant-temperature agitator, during liquid boiling, rapidly
The trisodium citrate through micro porous filtration 1% is added, is stirred.Liquid was changed into purple in 2 minutes, and being changed into quickly is red,
Fluidized state is kept, continues to boil 15 minutes, powers off, be cooled to room temperature, detection system is carried out with ultraviolet-uisible spectrophotometer
Standby sample, maximum absorption wavelength is between 526-522, the collaurum of preparation, in 4 DEG C of refrigerators.
3. colloid gold label monoclonal antibody.The collaurum 1mL prepared as stated above is in vitro added, adds 0.1M
K2CO3(potassium carbonate) adjusts pH value to optimum pH 7.4, stirs, adds the monoclonal antibody 4C12 of purifying, often pipe is final
Protein content be 18 μ g/mL, stir 15min；Then the BSA (bovine serum albumin(BSA)) that mass concentration is 10% is added, is made
BSA ultimate densities reach 1%, continue to stir 30min.Then 1h is centrifuged with 12000rpm on high speed freezing centrifuge；Centrifugation
Terminate, carefully suction out supernatant, precipitate 0.02M borate buffer solution ((0.02M borate+5%BSA+20% sucrose+
0.01%NaN3(three nitrogen sodium)) it is resuspended, produce gold mark monoclonal antibody.
4. with pretreatment fluid, (pretreatment fluid is respectively：PH7.2 0.01M PBS (phosphate buffer)+1%BSA+
The triton -100 of+0.1% Tween-20 of 2.5% sucrose+0.05%) handle gold standard pad and sample pad.
5. by the monoclonal antibody 4C12 of the anti-Tilapia mossambica Streptococcusagalactiae of the colloid gold label prepared as stated above with
The amount specking for often padding 10.0 μ L is coated with label pad, then by the drying of 37 DEG C of label pad, be sealed.
6. by the monoclonal antibody 3A9 coating buffers of anti-Tilapia mossambica Streptococcusagalactiae antigen, (pH7.2 0.01M PBS delay
The sucrose of fliud flushing+0.8%NaCl (sodium chloride)+2%) 0.6mg/mL concentration is diluted to, it is coated with by 2.0 μ L amount to coated film
Detection line (T lines)；Goat anti-mouse immunoglobulin antibody is diluted to 2.0mg/mL concentration with coating buffer (being same as above), by 2.0 μ
L amount is coated with to the nature controlling line of coated film (C), 37 DEG C of dryings, is sealed
It is pasted onto 7. sample pad, label pad, coated film, adsorptive pads are overlapped successively on PVC support lining plates, test strips are
Into.
8. detection sample can be serum or tissue homogenate, Tilapia mossambica Streptococcusagalactiae is carried out using test strips of the present invention
During detection, take liquid sample 60-80 μ L to be directly added drop-wise in sample pad, result is observed after 5-10min, T, nature controlling line (C) are aobvious
Color, represent positive；Only nature controlling line (C) develops the color, and represents negative；If only detection line (T lines) colour developing or no display band, is
Null result.
Compared with prior art, the invention has the advantages that：
Because test strips of the present invention contain identification antibody (the monoclonal antibody 4C12 of two kinds of Tilapia mossambica Streptococcusagalactiaes
With monoclonal antibody 3A9), and both monoclonal antibodies are that our Screening and Identifications from multiple monoclonal antibodies of preparation go out
Come, thus it is more preferable using reagent strip of the present invention detection Tilapia mossambica Streptococcusagalactiae specific effect.
The detection of Tilapia mossambica Streptococcusagalactiae is carried out using test strips of the present invention, compared with current detection method, without
Special detection instrument and equipment, there is the advantages that testing cost is low, easy to operate, quick, be obtained with 5 to 10 minutes detecting
As a result.
Compared with current detection method, the detection of Tilapia mossambica Streptococcusagalactiae is carried out using test strips of the present invention, without
Professional and technical personnel operates can and completes test, has use range wide, can especially carry out Site Detection use, is adapted to
Vast grass-roots unit's large-scale use.
Compared with conventional indirect enzyme-linked immunosorbent assay, Tilapia mossambica agalasisa hammer is carried out using test strips of the present invention
The detection of bacterium, it can directly make the judgement whether Tilapia mossambica infects Streptococcusagalactiae, it may also be used for clinical Tilapia mossambica agalasisa hammer
The evaluation of bacterium disease therapeutic effect.
Brief description of the drawings
Fig. 1 is the structural representation of the test strip of the present invention.
In figure：Support lining plate 1, sample pad 2, label pad 3, coated film 4, adsorptive pads 5, nature controlling line C, detection line T.
Embodiment
Below in conjunction with drawings and Examples, the invention will be further described, and following examples are to be better understood from
The present invention, present disclosure and protection domain are not construed as limiting：
The structure of the test strip of the present invention is as shown in figure 1, Tilapia mossambica Streptococcusagalactiae immunochromatography quick detection is tried
Paper slip and preparation method thereof, the test strip are made up of supporting plate 1, sample pad 2, label pad 3, coated film 4, adsorptive pads 5.
Anti- Tilapia mossambica Streptococcusagalactiae monoclonal antibody 4C12 is coated with to the label pad 3.By anti-Tilapia mossambica agalasisa
Streptococcic monoclonal antibody 3A9 is coated with to the detection line T lines of the coated film 4.By goat anti-mouse immunoglobulin antibody
It is coated with to the nature controlling line C of the coated film 4.
The sample pad 2, label pad 3, coated film 4, adsorptive pads 2 are overlapped successively and are pasted onto in supporting plate 1, Tilapia mossambica
Streptococcusagalactiae immune chromatography rapid detecting test paper strip is prepared and completed.
The material of above-mentioned sample pad 2 and label pad 3 is glass fibre membrane, and the material of above-mentioned coated film 4 is nitrocellulose
Film, the material of above-mentioned adsorptive pads 5 is crude fibre blotting paper, and the test strips supporting plate 1 is the polyvinyl chloride panel not absorbed water.
The anti-Tilapia mossambica Streptococcusagalactiae monoclonal antibody 4C12 monoclonal antibodies of above-mentioned colloid gold label, which are used to be coated with, to be marked
The amount of note pad is often to pad 5 μ L-30 μ L.
Coating concentration of the 3A9 monoclonal antibodies of above-mentioned anti-Tilapia mossambica Streptococcusagalactiae antigen in detection line (T lines) is
0.6mg/mL, package amount are often pad 1-5 μ L/cm2；Coating of the goat anti-mouse immunoglobulin antibody on nature controlling line (C)
Concentration is 2.0mg/mL, and package amount is often pad 1-5 μ L/cm2。
The material source and preparation method being related in embodiment 1-4 are as follows：
1. with above two hybridoma, (preserving number is according to a conventional method：CCTCC NO:C2014248：Preserving number is:
CCTCC NO:C2014244 mouse ascites monoclonal antibody 4C12 and 3A9) are prepared respectively.The anti-Tilapia mossambica Streptococcusagalactiae
The monoclonal antibody 4C12 and 3A9 of antigen are essentially protein, can specific recognition Tilapia mossambica Streptococcusagalactiae.
2. the collaurum of 20-40nm size particles is prepared using sodium citrate according to a conventional method.Specific method is as follows：
0.01% secondary gold chloride is added in 250ml conical flask, is heated on electromagnetic constant-temperature agitator, during liquid boiling, rapidly
The trisodium citrate through micro porous filtration 1% is added, is stirred.Liquid was changed into purple in 2 minutes, and being changed into quickly is red,
Fluidized state is kept, continues to boil 15 minutes, powers off, be cooled to room temperature, detection system is carried out with ultraviolet-uisible spectrophotometer
Standby sample, maximum absorption wavelength is between 526-522, the collaurum of preparation, in 4 DEG C of refrigerators.
3. colloid gold label monoclonal antibody.The collaurum 1mL prepared as stated above is in vitro added, adds 0.1M
K2CO3(potassium carbonate) adjusts pH value to optimum pH 7.4, stirs, adds the monoclonal antibody 4C12 of purifying, often pipe is final
Protein content be 18 μ g/mL, stir 15min；Then the BSA (bovine serum albumin(BSA)) that mass concentration is 10% is added, is made
BSA ultimate densities reach 1%, continue to stir 30min.Then 1h is centrifuged with 12000rpm on high speed freezing centrifuge；Centrifugation
Terminate, carefully suction out supernatant, precipitate 0.02M borate buffer solution ((0.02M borate+5%BSA+20% sucrose+
0.01%NaN3(three nitrogen sodium)) it is resuspended, produce gold mark monoclonal antibody.
4. with pretreatment fluid, (pretreatment fluid is respectively：Ph7.2 0.01M PBS+1%BSA+2.5% sucrose+0.1%
The triton of Tween-20+0.05% -100) handle gold standard pad and sample pad
5. by the monoclonal antibody of the anti-Tilapia mossambica Streptococcusagalactiae antigen of the colloid gold label prepared as stated above
4C12, be coated with 10.0 μ L of every pad amount specking in label pad, then by the drying of 37 DEG C of label pad, be sealed.
6. by the monoclonal antibody 3A9 coating buffers of anti-Tilapia mossambica Streptococcusagalactiae antigen, (pH7.2 0.01M PBS delay
The sucrose of fliud flushing+0.8%NaCl (sodium chloride)+2%) 0.6mg/mL concentration is diluted to, it is coated with by 2.0 μ L amount to coated film
Detection line (T lines)；Goat anti-mouse immunoglobulin antibody is diluted to 2.0mg/mL concentration with coating buffer (being same as above), by 2.0 μ
L amount is coated with to the nature controlling line of coated film (C), 37 DEG C of dryings, is sealed.
7. sample pad, label pad, coated film, adsorptive pads are overlapped be pasted onto on support lining plate successively, that is, obtain this Rofe
Fish Streptococcusagalactiae immune chromatography rapid detecting test paper strip.
8. detection sample can be serum or tissue fluid, Tilapia mossambica Streptococcusagalactiae detection is carried out using test strips of the present invention
When, taking liquid sample 60-80 μ L to be directly added drop-wise in sample pad, result is observed after 5-10min, T, nature controlling line (C) develop the color,
Represent positive；Only nature controlling line (C) develops the color, and represents negative；If only detection line (T lines) colour developing or no display band, is nothing
Imitate result.
In above-mentioned steps, BCA method protein concentrations kit is purchased from the green skies biotech firm in Nanjing；
Goat anti-mouse immunoglobulin antibody is the century limited public affairs of biotechnology purchased from Beijing health
BSA (bovine serum albumin(BSA)) is Sigma Products
Chemical reagent is purchased from traditional Chinese medicines company, is AR；
The 0.01mol/L PBS cushioning liquid (dilution) of PH 7.2 is prepared：
Na2HPO412H2O 0.2579g are weighed, NaH2PO42H2O 0.0437g add distilled water to 100mL adjustment
To 7.2,0.22 μm of filtering with microporous membrane, 4 DEG C save backup pH value.
Embodiment 1
By the anti-Tilapia mossambica Streptococcusagalactiae monoclonal antibody 4C12 of the colloid gold label prepared as stated above with every pad
10.0 μ L amount specking is coated with label pad, then by the drying of 37 DEG C of label pad, be sealed.
The monoclonal antibody 3A9 of anti-Tilapia mossambica Streptococcusagalactiae is diluted to 0.6mg/mL concentration, by 2.0 μ L amount
It is coated with to the detection line (T lines) of coated film；Goat anti-mouse immunoglobulin antibody is diluted to 2.0mg/mL concentration, by 2 μ L
Amount be coated with to the nature controlling line of coated film (C), 37 DEG C of dryings, be sealed.
Sample pad, label pad, coated film, adsorptive pads are overlapped successively and are pasted onto on PVC support lining plates, Tilapia mossambica agalasisa chain
Coccus immune chromatography rapid detecting test paper strip is prepared and completed.
Tilapia mossambica Streptococcusagalactiae immune chromatography rapid detecting test paper strip of the present invention, detection sample serum, tissue are even
Slurries etc., when carrying out the detection of Tilapia mossambica Streptococcusagalactiae using test strips of the present invention, liquid sample liquid 60-80 μ L are taken directly to be added dropwise
Onto sample pad, the target substance in sample liquid is Tilapia mossambica Streptococcusagalactiae, along test strips under fibrous layer capillarity
From be stained with liquid end to handheld terminal move, when it moves through label pad, Tilapia mossambica Streptococcusagalactiae i.e. with it is coated in label pad
The anti-Tilapia mossambica Streptococcusagalactiae monoclonal antibody 4C12 of colloid gold label is combined, and continues to move to coated film fibrous layer, when
When this antigen-antibody conjugate moves through coated film T (detection) line, with coated anti-Tilapia mossambica agalasisa in detection line (T lines)
Streptococcic monoclonal antibody 3A9 specific bindings, cause collaurum to be assembled in detection line, detection line is developed the color, remaining glue
The anti-Tilapia mossambica Streptococcusagalactiae monoclonal antibody 4C12 of body gold mark continues to migrate forward, when moving to nature controlling line (C) due to
The anti-Tilapia mossambica Streptococcusagalactiae monoclonal antibody 4C12 of colloid gold label can exempt from coated sheep anti-Mouse on nature controlling line (C)
Epidemic disease globulin antibody is specifically bound, and therefore, collaurum is assembled on nature controlling line (C), control line is developed the color, i.e. detection line (T
Line) and nature controlling line (C) develop the color simultaneously, show sample liquid to be checked for the positive.If sample liquid to be checked does not have Tilapia mossambica agalasisa hammer
Bacterium, then, when sample liquid to be checked is migrated to detection line (T lines), sample liquid to be checked not with coated colloid in detection line (T lines)
The composition that the anti-Tilapia mossambica Streptococcusagalactiae monoclonal antibody 4C12 of gold mark is combined, so as to cause collaurum not to be gathered in inspection
On survey line (T), therefore detection line (T) will not develop the color, but still be developed the color on nature controlling line (C), i.e., now only have nature controlling line (C)
Colour developing, show sample liquid to be checked for feminine gender.Testing result can judge in 5-10 clocks.
Embodiment 2
The specificity verification of Tilapia mossambica Streptococcusagalactiae is determined.According to the detection Tilapia mossambica agalasisa chain described in embodiment 1
Coccus immune chromatography rapid detecting test paper strip preparation method prepares test strips.
Specific test is done with common Streptococcus iniae, Vibrio anguillarum, thermophilic aqueous vapor pseudomonas bacillus, Pseudomonas fluorescence,
And do blank control.
Testing result shows：Only when detecting Streptococcusagalactiae, detection line (T lines) and nature controlling line (C) in test strips
There was only nature controlling line (C) colour developing when just developing the color and be, and detecting other antigens simultaneously.
Prove to use described detection Streptococcusagalactiae immune chromatography rapid detecting test paper strip detection Streptococcus iniae, eel arc
Bacterium, thermophilic aqueous vapor pseudomonas bacillus, during Pseudomonas fluorescence, cross reaction does not occur with these antigens, illustrate described detection nothing
Streptococcus lactis immune chromatography rapid detecting test paper strip has good detection specificity.
Embodiment 3
Checking to the detection sensitivity of Streptococcusagalactiae determines.Exempt from according to the detection Streptococcusagalactiae described in embodiment 1
Epidemic disease chromatography Rapid detection test strip preparation method prepares test strips.If 1.5 × 107、1.5×106、 1.5×105With 1.5 ×
1044 Streptococcusagalactiae concentration, and blank control is set, carry out sensitivity tests.As a result show：
Test strips of the present invention can detect that the bacteria concentration of Streptococcusagalactiae is 1.5 × 105。
Embodiment 4
The detection of experimental infection sample Streptococcusagalactiae.According to the detection Streptococcusagalactiae immunochromatography described in embodiment 1
Rapid detection test strip preparation method prepares test strips.With 1.5 × 109Streptococcusagalactiae cell concentration, it is difficult to understand per tail through abdominal cavity
Buddhist nun Tilapia mossambica (30 grammes per square metre) is inoculated with 0.3mL, and the various tissue samples that Tilapia mossambica is gathered after symptom occurs in Tilapia mossambica are checked,
Tissue sample is homogenized, and then various tissue aspiration tissue homogenate supernatant 60-80 μ L samples liquid are directly added drop-wise to each test paper
In bar sample pad, result is observed after 5-10min, the results showed that：Brain tissue, head-kidney tissue, liver organization, spleen tissue, rear kidney
Tissue, intestinal tissue, T, the nature controlling line (C) of various tissue detections develop the color, and represent positive.Blank control only has nature controlling line (C)
Colour developing, represent negative.Illustrate respectively to organize detect Streptococcusagalactiae from the Tilapia mossambica of infection.
Claims (4)
1. Tilapia mossambica Streptococcusagalactiae immune chromatography rapid detecting test paper strip and preparation method thereof, it is characterised in that：The detection
Test strips are made up of supporting plate (1), sample pad (2), label pad (3), coated film (4), adsorptive pads (5)；By anti-Tilapia mossambica agalasisa
Streptococcus monoclonal antibody 4C12 is coated with to the label pad (3)；By the monoclonal antibody 3A9 of anti-Tilapia mossambica Streptococcusagalactiae
In coating to the detection line (T lines) of the coated film (4)；Goat anti-mouse immunoglobulin antibody is coated with to the coated film
(4) on nature controlling line (C)；The sample pad (2), label pad (3), coated film (4), adsorptive pads (2) are overlapped successively and are pasted onto
In supporting plate (1), Tilapia mossambica Streptococcusagalactiae immune chromatography rapid detecting test paper strip is prepared and completed.
2. a kind of Tilapia mossambica Streptococcusagalactiae immune chromatography rapid detecting test paper strip according to claim 1, its feature exist
In：The material of the sample pad (2) and label pad (3) is glass fibre membrane, and the material of the coated film (4) is nitrocellulose
Film, the material of the adsorptive pads (5) is crude fibre blotting paper, and the test strips supporting plate (1) is the polyvinyl chloride panel not absorbed water.
3. a kind of Tilapia mossambica Streptococcusagalactiae immune chromatography rapid detecting test paper strip according to claim 3, its feature exist
In：The anti-Tilapia mossambica Streptococcusagalactiae monoclonal antibody 4C12 monoclonal antibodies of the colloid gold label are used to be coated with label pad
Measure often to pad 5 μ L-30 μ L.
4. a kind of Tilapia mossambica Streptococcusagalactiae immune chromatography rapid detecting test paper strip according to claim 1-3, its feature
It is, the coating concentration of the 3A9 monoclonal antibodies of the anti-Tilapia mossambica Streptococcusagalactiae antigen in detection line (T lines) is
0.6mg/mL, package amount are often pad 1-5 μ L/cm2；Coating of the goat anti-mouse immunoglobulin antibody on nature controlling line (C)
Concentration is 2.0mg/mL, and package amount is often pad 1-5 μ L/cm2。
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