Pig Epidemic Diarrhea Virus Antibody (PEDV-Ab) ELISA Kit
Référence AE64505PI
Conditionnement : 96Tests
Marque : Abebio
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Téléphone : +1 850 650 7790
Reagent Preparation
Results demonstration
Assay Procedure Summary
Certificate
Product Details
| Species Reactivity | Pig (Sus scrofa; Porcine) |
| UniProt | N/A |
| Abbreviation | PEDV-Ab |
| Alternative Names | N/A |
| Range | Request Information |
| Sensitivity | Request Information |
| Sample Type | Serum, Plasma, Other biological fluids |
| Detection Method | Competitive ELISA |
| Analysis Method | Qualitative |
| Assay Duration | 1-3h |
| Sample Volume | 1-200 μL |
| Detection Wavelengt | 450 nm |
Test principle
The microtiter plate provided in this kit has been pre-coated with PEDV. Samples are then added to the appropriate microtiter plate wells with a Horseradish Peroxidase (HRP)-conjugated Anti-Human antibody and incubated. Then substrate solutions are added to each well. Only those wells that contain PEDV-Ab and enzyme-conjugated Anti-Human antibody will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm.
Product Overview
Porcine epidemic diarrhea virus (PED virus or PEDV) is a coronavirus that infects the cells lining the small intestine of a pig, causing porcine epidemic diarrhoea, a condition of severe diarrhea and dehydration. Older hogs mostly get sick and lose weight after being infected, whereas newborn piglets usually die within five days of contracting the virus. PEDV cannot be transmitted to humans, nor contaminate the human food supply.
PEDV has a substantial economic burden given that it is highly infectious, resulting in significant morbidity and mortality in piglets. Morbidity and mortality rates were lower for vaccinated herds than for nonvaccinated herds, which suggests the emergence of a new PEDV field strain(s) for which the current vaccine, based on the CV777 strain, was partially protective. Consumers are likely to feel the effects of the viral disease in the form of higher for pork products.
Components
Reagents | Quantity | Reagents | Quantity |
Assay plate (96 Wells) | 1 | Instruction manual | 1 |
Standard (lyophilized) | 2 | Sample Diluent | 1 x 20 mL |
Biotin-Conjugate (concentrate 100 x) | 1 x 120 μL | Biotin-Conjugate Diluent | 1 x 12 mL |
Streptavidin-HRP (concentrate 100 x) | 1 x 120 μL | Streptavidin-HRP Diluent | 1 x 12 mL |
Wash Buffer (concentrate 25 x) | 1 x 20 mL | Substrate Solution | 1 x 10 mL |
Stop Solution | 1 x 6 mL | Adhesive Films | 4 |
Specificity
This assay has high sensitivity and excellent specificity for detection of PEDV-Ab. No significant cross-reactivity or interference between PEDV-Ab and analogues was observed.
Recovery
Matrices listed below were spiked with certain level of recombinant PEDV-Ab and the recovery rates were calculated by comparing the measured value to the expected amount of PEDV-Ab in samples.
Precision
Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in forty separate assays to assess inter-assay precision. CV (%) = SD/meanX100 Intra-Assay: CV<8% Inter-Assay: CV<12%
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of PEDV-Ab and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Stability
The stability of ELISA kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. The loss rate was determined by accelerated thermal degradation test. Keep the kit at 37°C for 4 and 7 days, and compare O.D.values of the kit kept at 37°C with that of at recommended temperature. (referring from China Biological Products Standard, which was calculated by the Arrhenius equation. For ELISA kit, 4 days storage at 37°C can be considered as 6 months at 2 - 8°C, which means 7 days at 37°C equaling 12 months at 2 - 8°C).
Sample collection and storage
Serum: Use a serum separator tube (SST) and allow samples to clot for two hours at room temperature or overnight at 2 - 8°C before centrifugation for 15 minutes at 1000 × g. Remove serum and assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles. Plasma: Collect plasma using EDTA, or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000 × g at 2 - 8°C within 30 minutes of collection. Assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles. Other biological fluids: Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.
Kits storage instructions
Store at 2-8°C. Please refer to Instruction Manual.