AU2021106008A4 - Kit for Detecting Antibody Against Foot-and-mouth Disease Virus 2C - Google Patents
Kit for Detecting Antibody Against Foot-and-mouth Disease Virus 2C Download PDFInfo
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- AU2021106008A4 AU2021106008A4 AU2021106008A AU2021106008A AU2021106008A4 AU 2021106008 A4 AU2021106008 A4 AU 2021106008A4 AU 2021106008 A AU2021106008 A AU 2021106008A AU 2021106008 A AU2021106008 A AU 2021106008A AU 2021106008 A4 AU2021106008 A4 AU 2021106008A4
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- Prior art keywords
- enzyme
- mouth disease
- disease virus
- foot
- kit
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- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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- 241000710198 Foot-and-mouth disease virus Species 0.000 title claims abstract description 31
- 102000004190 Enzymes Human genes 0.000 claims abstract description 45
- 108090000790 Enzymes Proteins 0.000 claims abstract description 45
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 8
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 8
- 210000002966 serum Anatomy 0.000 claims description 36
- 239000007788 liquid Substances 0.000 claims description 24
- 238000005406 washing Methods 0.000 claims description 21
- 239000003085 diluting agent Substances 0.000 claims description 11
- 239000000758 substrate Substances 0.000 claims description 11
- 239000008363 phosphate buffer Substances 0.000 claims description 10
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 claims description 8
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 8
- 238000000576 coating method Methods 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 6
- 239000011248 coating agent Substances 0.000 claims description 5
- 235000020183 skimmed milk Nutrition 0.000 claims description 5
- 241000283690 Bos taurus Species 0.000 claims description 4
- YRNWIFYIFSBPAU-UHFFFAOYSA-N 4-[4-(dimethylamino)phenyl]-n,n-dimethylaniline Chemical compound C1=CC(N(C)C)=CC=C1C1=CC=C(N(C)C)C=C1 YRNWIFYIFSBPAU-UHFFFAOYSA-N 0.000 claims description 3
- 229920001213 Polysorbate 20 Polymers 0.000 claims description 3
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 claims description 3
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 claims description 3
- 238000000034 method Methods 0.000 abstract description 9
- 238000001514 detection method Methods 0.000 abstract description 7
- 238000002965 ELISA Methods 0.000 abstract description 6
- 230000035945 sensitivity Effects 0.000 abstract description 3
- 239000000243 solution Substances 0.000 description 26
- 241000700605 Viruses Species 0.000 description 16
- 241001465754 Metazoa Species 0.000 description 8
- 201000010099 disease Diseases 0.000 description 7
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 7
- 208000007212 Foot-and-Mouth Disease Diseases 0.000 description 5
- 238000011534 incubation Methods 0.000 description 5
- 238000011161 development Methods 0.000 description 4
- 238000001035 drying Methods 0.000 description 4
- 208000030194 mouth disease Diseases 0.000 description 4
- 229910019142 PO4 Inorganic materials 0.000 description 3
- 239000000427 antigen Substances 0.000 description 3
- 102000036639 antigens Human genes 0.000 description 3
- 108091007433 antigens Proteins 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 3
- 239000010452 phosphate Substances 0.000 description 3
- 241000710777 Classical swine fever virus Species 0.000 description 2
- 241000702619 Porcine parvovirus Species 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 208000032625 disorder of ear Diseases 0.000 description 2
- 239000003547 immunosorbent Substances 0.000 description 2
- 235000013336 milk Nutrition 0.000 description 2
- 239000008267 milk Substances 0.000 description 2
- 210000004080 milk Anatomy 0.000 description 2
- 238000011017 operating method Methods 0.000 description 2
- 239000012089 stop solution Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- 241001533384 Circovirus Species 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 241000710842 Japanese encephalitis virus Species 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 241000202347 Porcine circovirus Species 0.000 description 1
- 206010037660 Pyrexia Diseases 0.000 description 1
- 241000701093 Suid alphaherpesvirus 1 Species 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 241000282898 Sus scrofa Species 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 230000036528 appetite Effects 0.000 description 1
- 235000019789 appetite Nutrition 0.000 description 1
- 238000009360 aquaculture Methods 0.000 description 1
- 244000144974 aquaculture Species 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 239000013065 commercial product Substances 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 1
- 210000000003 hoof Anatomy 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 208000009305 pseudorabies Diseases 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
Classifications
-
- 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
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
-
- 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
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
- G01N33/535—Production of labelled immunochemicals with enzyme label or co-enzymes, co-factors, enzyme inhibitors or enzyme substrates
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/38—Diluting, dispersing or mixing samples
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
- G01N2333/08—RNA viruses
- G01N2333/085—Picornaviridae, e.g. coxsackie virus, echovirus, enterovirus
- G01N2333/09—Foot-and-mouth disease virus
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2469/00—Immunoassays for the detection of microorganisms
- G01N2469/20—Detection of antibodies in sample from host which are directed against antigens from microorganisms
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/20—Dermatological disorders
Abstract
The invention provides a kit for detecting the antibody against foot-and-mouth
disease virus 2C, which relates to the technical field of enzyme-linked immunosorbent
assay. The enzyme label plate is coated by foot-and-mouth disease virus 2C protein.
The kit provided by the invention has the advantages of high efficiency, sensitivity,
specificity and repeatability for detecting the antibody against foot-and-mouth disease
virus 2C. The method is simple, rapid and low in cost, can be visually detected at room
temperature, is suitable for popularization in clinical application, and provides a reliable
technical means for rapid detection of the antibody against foot-and-mouth disease
virus 2C.
Description
Kit for Detecting Antibody Against Foot-and-mouth Disease Virus 2C
The invention relates to the technical field of enzyme-linked immunosorbent
assay, in particular to a kit for detecting the antibody against foot-and-mouth disease
virus 2C.
Foot-and-mouth disease is an acute, hot and highly contagious disease caused by
foot-and-mouth disease virus (FMDV), which mainly affects cloven-hoofed animals
and often spreads in sheep, cattle, pigs and other animals. Blisters appeared on the lips
and hoofs of susceptible animals at the time of onset, accompanied by fever, loss of
appetite and other symptoms, and the weight and milk yield of sick animals decreased
greatly. Foot-and-mouth disease virus is easy to spread through the air, with strong
infectivity and rapid epidemic. Susceptible animals will also die under the condition of
weak resistance, causing huge economic losses to farmers and seriously hindering the
development of aquaculture.
At present, the diagnosis method of antibody against foot-and-mouth disease virus
2C and the evaluation method of vaccine immune effect can only be completed in
laboratory, because the operation process can not be separated from laboratory
instruments and equipment, so it is not suitable for grass-roots detection.
The purpose of the present invention is to provide a kit for detecting the antibody
against foot-and-mouth disease virus 2C, which is suitable for grass-roots detection
without using laboratory instruments and equipment.
The invention provides a kit for detecting the antibody against foot-and-mouth
disease virus 2C, which comprises an enzyme-labeled plate and an enzyme-labeled
secondary antibody;
The enzyme label plate is coated by foot-and-mouth disease virus 2C protein.
Preferably, the coating concentration of the foot-and-mouth disease virus 2C
protein is 1-2 [g/mL.
Preferably, the kit further comprises washing liquid, serum diluent, substrate color
developing liquid, stopping liquid, positive serum and negative serum.
Preferably, the washing liquid comprises phosphate buffer containing Tween-20
with mass fraction of 0.03-0.08%, and the pH value of the phosphate buffer is 7.2.
Preferably, the serum diluent comprises phosphate buffer, which contains
skimmed milk powder, the concentration of which is 45-55 g/L, and the pH value of
the phosphate buffer is 7.2.
Preferably, the substrate color developing solution comprises TMB
(Tetramethylbenzidine) solution.
Preferably, the termination solution comprises SDS (Sodium dodecyl sulfate)
solution or sulfuric acid solution.
Preferably, the enzyme-labeled secondary antibody comprises a porcine enzyme
labeled secondary antibody or a bovine enzyme-labeled secondary antibody.
The invention provides a kit for detecting the antibody against foot-and-mouth
disease virus 2C, which comprises an enzyme-labeled plate and an enzyme-labeled
secondary antibody; the enzyme label plate is coated by foot-and-mouth disease virus
2C protein. In the present invention, when an animal is infected with foot-and-mouth
disease virus, 2C antibody is produced, and the detection purpose is achieved by
detecting the antibody against 2C.The results of the embodiment of the invention show
that the kit provided by the invention has the advantages of high efficiency, good
sensitivity, specificity and repeatability in detecting the antibody against foot-and
mouth disease virus 2C, and the method is simple and convenient to operate, fast and
low in cost, can be visually detected at room temperature, is suitable for popularization
in clinical application, and provides a reliable technical means for rapid detection of the
antibody against foot-and-mouth disease virus 2C.
Figure 1 shows the detection results of the kit provided by the invention, in which,
from left to right:
1 and 2 are foot-and-mouth disease virus serum; 3. Antibodies against porcine
epidemic diarrhea virus; 4. Classical swine fever virus; 5. Antibody against porcine
circovirus; 6. Antibody against porcine parvovirus; 7. Antibody against pseudorabies
virus; 8. Antibody against blue ear disease virus.
The invention provides a kit for detecting the antibody against foot-and-mouth
disease virus 2C, which comprises an enzyme-labeled plate and an enzyme-labeled secondary antibody; the enzyme label plate is coated by foot-and-mouth disease virus
2C protein.
In the present invention, the enzyme-labeled plate is coated with foot-and-mouth
disease virus 2C protein,, and the concentration of the coating is preferably 1-2 [g/mL.
According to the invention, the coating method is not particularly limited, and
conventional coating can be adopted.
In the present invention, the enzyme label plate is preferably sealed with a skim
milk powder solution with a concentration of 50 g/L.
In the present invention, the enzyme-labeled secondary antibody preferably
comprises a pig's enzyme-labeled secondary antibody or a cow's enzyme-labeled
secondary antibody, which is preferably a commercial product.
In the present invention, the kit also preferably includes washing solution, serum
diluent, substrate color developing solution, stopping solution, positive serum and
negative serum.
In the present invention, the washing liquid preferably comprises phosphate
buffer, which contains Tween-20 with a mass fraction of 0.03-0.08%, more preferably
0.05%; The pH value of the phosphate buffer solution is 7.2.
In the present invention, the serum diluent preferably comprises phosphate buffer,
and the buffer contains skim milk powder, and the concentration of the skim milk
powder is preferably 45-55g/L, more preferably 50g/L; The pH value of the phosphate
buffer solution is preferably 7.2.
In the present invention, the substrate color developing solution preferably
comprises TMB solution, and the concentration of the TMB solution is not particularly
limited in the present invention, and the required TMB concentration can be used when
conventional substrates are used for color development.
In the present invention, the termination solution preferably comprises an SDS
solution or a sulfuric acid solution. According to the invention, the concentration of the
SDS solution or the sulfuric acid solution is not particularly limited, and the
concentration required for conventionally stopping the enzyme-linked immunosorbent
assay can be adopted.
In the present invention, the positive serum is preferably the serum infected with
foot-and-mouth disease, and the negative serum is preferably the serum of healthy
animals.
In the present invention, the method for detecting the antibody against foot-and
mouth disease virus 2C by the kit preferably comprises the following steps:
1) Diluting a serum sample with the serum diluent at a volume ratio of 1:40 to
obtain diluted serum;
2) Adding the diluted serum obtained in step 1) into the hole of the enzyme-labeled
plate, incubating at 20-25C for 50-70 min, discarding the liquid in the hole, washing
with washing liquid, and pat-drying to obtain a first pat-dried enzyme-labeled plate;
3) Adding diluted enzyme labeled secondary antibody into the hole of the first dry
enzyme labeled plate obtained in step 2), incubating at 20-25 °C for 50-70 min, discarding the liquid in the hole, washing with washing liquid, and drying to obtain a second dry enzyme labeled plate;
4) Adding substrate chromogenic solution into the hole of the second dry ELISA
plate obtained in step 3), and chromogenic at 20-25C for 15-25 min, when the color
in the hole is blue, the antibody against foot-and-mouth disease virus 2C is detected,
and when the color in the hole is colorless, the antibody against foot-and-mouth disease
virus 2C is not detected.
According to the invention, the serum sample is diluted by the serum diluent in a
volume ratio of 1:40 to obtain diluted serum.
The method comprises the following steps: adding the obtained diluted serum into
the hole of the enzyme-labeled plate, incubating at 20-25C for 50-70 min, discarding
the liquid in the hole, washing with washing liquid, and pat-drying to obtain the first
pat-dried enzyme-labeled plate.
In the present invention, the amount of diluted serum added into the hole of the
enzyme-labeled plate is preferably 100 [L.
In the invention, diluted serum is added into the hole of the enzyme-labeled plate
and then incubated at 20-25°C, preferably 21-24°C, more preferably 22-23°C; The
incubation time is 50-70 min, preferably 55-65 min, and most preferably 60min.
In the present invention, the number of times of washing is preferably 3 times.
The method comprises the following steps of: adding diluted enzyme-labeled
secondary antibody into the hole of the obtained first dry enzyme-labeled plate, incubating it at 20-25C for 50-70 min, then discarding the liquid in the hole, washing with washing liquid, and drying to obtain the second dry enzyme-labeled plate.
In the present invention, the amount of diluted enzyme labeled secondary antibody
added into the hole of the first enzyme labeled plate is preferably 100. m. In the present
invention, the enzyme-labeled secondary antibody is preferably diluted with phosphate
buffer solution, and the dilution multiple is preferably 20,000 times.
In the present invention, the diluted ELISA secondary antibody is added into the
hole of the first dry ELISA plate for incubation, and the incubation temperature is 20
°C, preferably 21-24°C, more preferably 22-23°C; The incubation time is 50-70 min,
preferably 55-65 min, and most preferably 60 min.
In the present invention, the number of times of washing is preferably 3 times.
According to the invention, the substrate color developing solution is added into
the hole of the obtained second beat dry enzyme label plate, and the color is developed
for 15-25 min at 20-25°C. When the color in the hole is blue, the antibody against foot
and-mouth disease virus 2C is detected, and when the color in the hole is colorless, the
antibody against foot-and-mouth disease virus 2C is not detected.
In the present invention, the addition amount of the substrate color developing
solution is preferably 100 L/hole. In the present invention, the color developing
temperature is preferably 21-24°C, more preferably 22-23°C; the color development
time is preferably 18-23 min, preferably is 20min.
The kit for detecting the antibody against foot-and-mouth disease virus 2C
according to the present invention will be further illustrated in detail with specific examples. The technical scheme of the present invention includes but is not limited to the following examples.
Example 1
1. Preparation of sample diluent, washing solution and stop solution: the coating
solution is 0.05M of carbonate buffer: 1.59g ofNa2CO3, 2.93g ofNaHCO3, and distilled
water is added to make the volume constant to 1000 mL, and the pH value is 9.6;
Sample diluent is washing liquid containing 50 g/L skimmed milk powder;
The washing solution is 0.01MpH 7.2 phosphate buffer containing 0. 05% Tween
, and its formulation is: 8.5 g of NaCl, 0.356g of NaH2PO 4 -2H2O, 2.772g of
NaH2PO 4 -12H 2 0, which are mixed, then dissolved in distilled water and fixed to
1000mL. Termination solution: prepare 1% SDS solution.
2. Determination of optimal concentration of antigen and optimal dilution of
serum:
The results showed that when the dilution of antigen was 2.0 g/mL per hole, the
serum was diluted by 1:40 times, and the enzyme-labeled secondary antibody was
diluted by 1:20000 times.
3. Optimization of action time
The best blocking time of antigen is 20min, the reaction time of serum is lh, the
best incubation time of enzyme-labeled secondary antibody is 40min, and the best color
development time is 20min.
4. Determination of operating procedures
According to the best operating conditions determined above, the determined
operating procedure is as follows: take out the coated and sealed ELISA strip, dilute
the serum to be detected by 1:40 times with serum diluent, add 100 L to each hole of
the ELISA plate, add one hole to each hole, and add two holes to the negative and
positive serum of type A foot-and-mouth disease virus (the positive and negative serum
are respectively infected with foot-and-mouth disease and healthy animal serum,
diluted 100 times with PBS), at room temperature for 1 h.
Discard the liquid in the hole, wash each hole with washing liquid for 3 times and
pat, and discard the liquid in the hole completely; add 100 L of enzyme-labeled
secondary antibody to each hole, and react at room temperature for 1 hour; Discard the
liquid in the hole, wash each hole with washing liquid for 3 times and pat, and discard
the liquid in the hole completely; After adding 100 L of TMB(3 ,3 ',5 5 '
tetramethylbenzidine) substrate color solution to each hole, develop color at room
temperature for 20 minutes in the dark, and add 100 L of SDS stop solution; observe
the color with eyes, blue is positive, colorless is negative.
Example 2
Specificity test of kit:
According to the method established in Example 1, 2C positive serum samples of
known classical swine fever virus, Japanese encephalitis virus, porcine circovirus,
porcine parvovirus, pseudorabies virus, blue ear disease virus and foot-and-mouth
disease virus were detected respectively, and the negative samples were colorless, while
the positive samples were blue. The results are shown in Figure 1.
According to fig. 1, the kit provided by the invention can accurately detect foot
and-mouth disease without cross reaction with other viruses while has specificity.
It can be concluded from the above embodiments that the kit provided by the
present invention has the advantages of high efficiency, sensitivity, specificity and
repeatability in detecting the antibody against foot-and-mouth disease virus 2C. The
method is simple, rapid and low in cost, can be visually detected at room temperature,
is suitable for popularization in clinical application, and provides a reliable technical
means for rapid detection of the antibody against foot-and-mouth disease virus 2C.
The above are only the preferred embodiments of the present invention, and it
should be pointed out that for ordinary people in the technical field, without departing
from the principle of the present invention, several improvements and embellishments
can be made, and these improvements and embellishments should also be regarded as
the protection scope of the present invention.
Claims (8)
1. A kit for detecting the antibody against foot-and-mouth disease virus 2C is
characterized by comprising an enzyme-labeled plate and an enzyme-labeled secondary
antibody;
The enzyme label plate is coated by foot-and-mouth disease virus 2C protein.
2. The kit according to claim 1 is characterized in that the coating concentration
of the foot-and-mouth disease virus 2C protein is 1-2 g/mL.
3. The kit according to claim 1 or 2 is characterized in that the kit further comprises
washing liquid, serum diluent, substrate color developing liquid, stopping liquid,
positive serum and negative serum.
4. The kit according to claim 3 is characterized in that the washing liquid
comprises phosphate buffer containing Tween-20 with mass fraction of 0.03-0.08%,
and the pH value of the phosphate buffer is 7.2.
5. The kit according to claim 3 is characterized in that the serum diluent comprises
phosphate buffer, which contains skimmed milk powder, the concentration of which is
-55 g/L, and the pH value of the phosphate buffer is 7.2.
6. The kit according to claim 3 is characterized in that the substrate color
developing solution comprises TMB (Tetramethylbenzidine) solution.
7. The kit according to claim 3 is characterized in that the termination solution
comprises SDS (Sodium dodecyl sulfate) solution or sulfuric acid solution.
8. The kit according to claim 1 is characterized in that the enzyme-labeled
secondary antibody comprises a porcine enzyme-labeled secondary antibody or a
bovine enzyme-labeled secondary antibody.
FIGURES 1/1
Figure 1
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| AU2021106008A AU2021106008A4 (en) | 2021-08-19 | 2021-08-19 | Kit for Detecting Antibody Against Foot-and-mouth Disease Virus 2C |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| AU2021106008A AU2021106008A4 (en) | 2021-08-19 | 2021-08-19 | Kit for Detecting Antibody Against Foot-and-mouth Disease Virus 2C |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| AU2021106008A4 true AU2021106008A4 (en) | 2021-10-28 |
Family
ID=78179611
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| AU2021106008A Ceased AU2021106008A4 (en) | 2021-08-19 | 2021-08-19 | Kit for Detecting Antibody Against Foot-and-mouth Disease Virus 2C |
Country Status (1)
| Country | Link |
|---|---|
| AU (1) | AU2021106008A4 (en) |
-
2021
- 2021-08-19 AU AU2021106008A patent/AU2021106008A4/en not_active Ceased
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