Comparison with the Pyro detect system
The Pyro Detect System is use according to the manufacture’s instruction as quantitative test with the exception that sample are not test at different dilutions. The Quantitative test is described by the procedures as method A. By using method A, a quantitative comparison of the samples with the standard endotoxin is possible. The stimulating agents are pipette out as 20 µl portion into 96-well plate under a horizontal flow bench. The spike and blank wells –the former is fill with stimulant and spike RPMI medium, with a total of 40µl RPMI-20µl RPMI are add to each well. The cryoblood vials (included in the kit) are thaw in a water bath for 1 minute and dilute immediately with 8 ml RPMI 1640 cell culture…show more content… Information is check for ordinary circulation by visual investigation and the Kolmogorov-Smirnov test. Some information sets demonstrate a left-skewed appropriation and fizzled the typicality…show more content… This test uses the same species as the established in vivo test, but the same material and endpoint as the new in vitro test with human blood. Several steps were necessary to establish the rabbit whole blood test:
i) Production of recombinant rabbit IL-1β in E. coli (positive control for the ELISA, substance to immunise the animals in step two), ii) Immunisation of a sheep with the IL-1β antigen, iii) Immunisation of mice with IL-1β and production of monoclonal antibodies, iv) Establishment of a sandwich
ELISA (Enzyme Linked Immunosorbent Assay) with the antibodies and the antigen. These steps were successfully completed recombinant IL-1β was produced in a reliable quality and sufficient quantity.
Monoclonal (mice) and polyclonal (sheep) antibodies against rabbit IL-1β were isolated. The ELISA assay allows the quantitative determination of the endogenous rabbit fever signal IL-1β.
Comparison of three pyrogen tests Test: Rabbit LAL WBT
Pyrogens Bacteria gram-negative + + + Bacteria gram-positive + - + Fungi + - +
Applications Biologicals + - + Pharmaceuticals + + + Medical Devices - + + Air quality - (+) + Blood components - -