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Pyrodetect System: the Human(e) Alternative in Pyrogen Testing
Broad Pyrogen Profile - Easy Assay Procedure - Robust Application
The PyroDetect System is a revolutionary test for the detection and quantification of pyrogens based on the Monocyte-Activation Test (MAT).
The PyroDetect System uses an innate immune defense reaction. Monocytes activated by pyrogens produce cytokines that are detected in an immunological assay (ELISA) involving specific antibodies and an enzyme-mediated color change.
The PyroDetect System consists of three main steps, the cryoblood (or fresh blood) incubation, the IL-1ß ELISA, and the analysis.
The PyroDetect System uses cryo-blood (pooled and frozen human whole blood) for the blood incubation. Fresh blood with a time limit of up to 4 hours after donation may also be used. The test samples are mixed with the blood in the cell culture plate and are kept in an incubator over night at 37 °C and 5 % CO2 for temperature and pH-stabilization. If pyrogens are present in the sample, the monocytes of the blood will produce IL-1ß during the incubation.
For the detection of the IL-1ß the cryo-blood incubation mixture is transferred into an ELISA microplate coated with an antibody specific for IL-1ß. Interleukin molecules present in the culture supernatant are bound by the immobilized antibody. Then an enzyme-linked polyclonal antibody specific for IL-1ß is added. With the addition of the substrate a color reaction is started, which allows the detection of the bound IL-1ß in an ELISA reader.
The pyrogen concentration in the sample is then determined from the IL-1ß concentration via an endotoxin standard curve. The PyroDetect System offers a high level of safety using both a positive and a negative control. The robust application of the test was shown in an international validation coordinated by the European Centre for the Validation of Alternative Methods to Animal Experimentation (ECVAM) in 2005. Several published studies show the robustness and reliability of the test performance of the MAT. The reproducibility of the PyroDetect System was shown in several user applications.
The PyroDetect System has a large pyrogen profile in the microbial field. Tests with Lipoteichoic Acid (LTA) indicate the detection of cell wall components of gram-positive bacteria with the PyroDetect System. As expected, the LAL test shows a reaction only to highly concentrated LTA. This is due to traces of endotoxin in LTA preparations. Thus the PyroDetect System reduces the security risk of non-endotoxin pyrogens and is an effective addition to the LAL test.
For further information please visit our product website, download our product brochure or get in contact with us!
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