Product Details
| Species Reactivity |
Human (Homo sapiens) |
| UniProt |
|
| Abbreviation |
Lep-Ab-IgG |
| Alternative Names |
N/A |
| Range |
Request Information |
| Sensitivity |
Request Information |
| Sample Type |
Serum, Plasma, Other biological fluids |
| Detection Method |
Sandwich |
| Analysis Method |
Quantitive |
| Assay Duration |
1-4.5h |
| Sample Volume |
1-200 μL |
| Detection Wavelengt |
405 nm |
Test principle
The microtiter plate provided in this kit has been pre-coated with Lep-Ag. Samples are then added to the appropriate microtiter plate wells with anti-Human IgG alkaline phosphatase (AP) and incubated. Then substrate solutions are added to each well. Only those wells that contain Lep-Ab-IgG and enzyme-conjugated Human IgG 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 405 nm ± 2 nm.
Product Overview
Leptospira (Greek leptos, fine, thin and Latin spira, coil) is a genus of spirochaete bacteria, including a small number of pathogenic and saprophytic species. Leptospira was first observed in 1907 in kidney tissue slices of a leptospirosis victim who was described as having died of yellow fever. Leptospira, together with the genera Leptonema and Turneria, is a member of the family Leptospiraceae. The genus Leptospira is divided into 20 species based on DNA hybridization studies.
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 Human Leptospira. No significant cross-reactivity or interference between Human Leptospira and analogues was observed.
Recovery
Matrices listed below were spiked with certain level of recombinant Human Leptospira and the recovery rates were calculated by comparing the measured value to the expected amount of Human Leptospira 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 Human Leptospira 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.
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