[Replacement of mouse and rat antibody production test; comparison of sensitivity between the in vitro and in vivo methods].
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 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.
Bootz
Frank
F
Institut für Labortierkunde Universität CH-Zürich, Switzerland. fobo@ltk.unizh.ch
Sieber
Isabella
I
D000826:Animal Testing Alternatives; D000818:Animals; D000917:Antibody Formation; D001483:Base Sequence; D017931:DNA Primers; D066298:In Vitro Techniques; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D017931:DNA Primers