Auleon Biologicals
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ELISA Kit for Detection of Goat and Sheep Antibody against EG95 Protein of Echinococcus (Hydatid)
Classification：
Details
[Veterinary drug name]
Echinococcus (hydatid) EG95 protein ELISA antibody detection kit
Trade name: None
英文名：ELISA Kit for Detection of Goat and Sheep Antibody against EG95 Protein of Echinococcus (Hydatid)
汉语拼音：Yang Jiqiuyou (Baochong) EG95 Danbai ELISA Kangti Jiance Shijihe
[Main composition and content]
| (1) | antigen coated plate | 96 well/plate x 2 plate/box |
| (2) | negative control serum | 50uL/tube x1 tube/box |
| (3) | positive control serum | 50uL/tube x1 tube/box |
| (4) | enzyme labeled secondary antibody | 0.2mL/tube x2 tube/box |
| (5) | Diluent | 30mL/bottle x2 bottle/box |
| (6) | 10x CONCENTRATED WASHER | 30mL/bottle x1 bottle/box |
| (7) | TMB substrate A solution | 10mL/bottle x1 bottle/box |
| (8) | TMB Substrate B Solution | 10mL/bottle x1 bottle/box |
| (9) | Stop fluid | 10mL/bottle x1 bottle/box |
| (10) | serum dilution plate | 96 well/plate x 2 plate/box |
| (11) | sealing plate membrane | 2 sheets/box |
| (12) | Instructions | 1 copy/box |
[Appearance]
The kit shall be sealed in good condition without damage, crack or leakage. Antigen coating plate, positive control serum, negative control serum, enzyme labeled secondary antibody, diluent, 10x concentrated washing solution, TMB substrate A solution, TMB substrate B solution, termination solution and other components are complete.
[Function]
It is used to detect antibodies to EG95 protein, the main protective antigen of Echinococcus granulosus (Echinococcus granulosus) in the serum of sheep and goats.
[Usage and Judgment]
1 Usage
1.1 sample preparation animal whole blood, after blood coagulation, 4000r/min centrifugation for 10 minutes, collect supernatant for standby; Can also collect blood, after coagulation naturally precipitate serum, suck out serum for standby. The serum is required to be clear without hemolysis.
1.2 the washing solution to prepare 10x concentrated washing solution. After returning to room temperature, shake well and dilute 10 times with deionized water for standby.
The 1.3 sample and the control substance are diluted on a serum dilution plate, and the serum to be tested and the negative and positive control serum are diluted in a volume of 1:50 times (recommend dilution method: 2uL of serum sample is added to 98uL of sample dilution).
1.4 operation steps
1.4.1 Add samples to take antigen-coated plates, add 50u diluent to each well, and then take 50uL of diluted serum to be tested and 50uL of control serum into corresponding wells. Among them, there are 2 wells of negative and positive control serum respectively, and gently shake and mix the samples in the wells (do not overflow).
1.4.2 Incubation was placed in a 37°C incubator for 30 minutes.
1.4.3 Discard the reaction liquid after washing, add 200uL of diluted washing liquid to each hole, soak at room temperature for 1 minute, use up the washing liquid, pat dry on absorbent paper, and repeat 3 times: or wash 3 times with an automatic plate washer.
1.4.4 Add enzyme-labeled secondary antibody Dilute the enzyme-labeled secondary antibody 1:100 with diluent (currently prepared for use), and add 100uL of diluted enzyme-labeled secondary antibody to each well.
1.4.5 Incubation was placed in a 37°C incubator for 30 minutes.
1.4.6 Washing is the same as 1.4.3.
1.4.7 Color development Add 100uL substrate working solution to each well (TMB substrate solution A and solution B are mixed in equal volume and are now prepared for current use), and color development reaction is carried out at room temperature for 10 minutes in the dark.
1.4.8 Add 50uL of termination solution to each hole and measure the results within 10 minutes.
1.4.9 Readings Read the 450nm wavelength absorbance of each well in a microplate reader.
2 Judgment
2.1 test establishment conditions positive control wells OD: values are> 1.20, negative control wells ODsom values are <0. 20.
The 2.2 results determined that the OD value of the serum pore to be tested was <0.30, which was judged as Echinococcus (hydatid) EG95 antibody negative. The OD:som value of the serum pore to be tested is ≥ 0.30, which is judged as Echinococcus (hydatid) EG95 antibody positive.
[Note]
(1) Except for the enzyme labeled secondary antibody, all kinds of reagents should be balanced to room temperature before use before use. If there is crystallization, it must be completely dissolved by heating in a water bath at 37 ℃. Mix the liquid reagent evenly before use. In order to ensure accurate and reliable results, the operating environment temperature should be controlled at 20±5 ℃.
(2) The micropipette should be calibrated regularly to avoid excessive errors affecting the test results.
(3) The quality of serum samples should be ensured, and severe hemolytic samples and fatty samples should be avoided. Avoid repeated freezing and thawing of clear samples; Serum samples should be stored at 2~8 ℃ for no more than 5 days, and should be kept below -20 ℃ for long-term storage. If turbidity or floccules appear in serum samples, supernatant should be taken for detection after centrifugation.
(4) Prevent mutual contamination of reagents and samples; the kit should be used within the validity period to avoid mixing of different batches of kits.
(5) The unused antigen-coated strips should be put back into the sealed bag together with the desiccant and stored at 2-8°C.
(6) During incubation, the reaction plates cannot be placed in a 37 ℃ incubator, so as not to cause uneven heating of each reaction plate.
(7)TMB substrate liquid A and liquid B should not be exposed to strong light and avoid contact with oxidants.
(8) The termination fluid has a corrosive effect and avoids contact with eyes and skin.
(9) Strictly follow the instructions and control the reaction time and temperature of each step.
(10) This kit cannot distinguish between antibodies produced by natural infection of Echinococcus granulosus and vaccine immunization: when this kit detects sheep serum infected with polycephalus, a positive rate of 7.69 (1/13) appears, therefore, the possibility of cross-reaction with sheep polycephalus antibody cannot be ruled out.
[Specification] 192 hole/box
[Storage and validity] 2~8 ℃, valid for 9 months.
[Approval No.] Veterinary Drug 230248948
[Production enterprise] Chongqing Aolong Biological Products Co., Ltd.
[Address] No.5, 4th Branch Road, Banqiao Industrial Park, Rongchang District, Chongqing
[Postcode] 402460
[Technical service] 023-46763511
[Sales hotline] 023-46761801
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