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Replacement of Mouse and Rat Antibody Production Test. Validation of Different Polymerase Chain Reaction Assays

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Frank Bootz, Isabella Sieber
Institut für Labortierkunde Universität CH-Zürich
Bacteria and viruses may be transmitted through contaminated biological materials such as transplantable tumors, cell lines, sera or other biological material.
Currently, the screening of biological material is being done using the mouse or rat antibody production (MAP/RAP) test (serological testing).
We decided to test and validate an alternative assay using polymerase chain reaction (PCR / RT-PCR) technology to detect viral contamination directly in biological material.
The aim of this study therefore is the validation of our new PCR assays and the comparison of PCR and MAP test.
For 6/14 viruses the conventional PCR seems to be more sensitive and more specific in detecting murine viruses. In 12/14 viruses the RT-PCR is more sensitive than the MAP-test. In 2/14 cases all detection methods have the same sensitivity. Furthermore, PCR screening clearly contributes to the principles of 3R as a replacement technique as it eliminates the need for using animals to screen for murine viruses in biological material.

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