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Entwicklung von spezifischen PCR-ELISA Nachweissystemen für Coxiella burnetii, Francisella tularensis und Orthopockenviren
(2001) Kohlhaußen, Simone; Böhm, Reinhard
In the present work specific systems on the basis of the PCR were developed for the detection of Coxiella burnetii, Francisella tularensis and Orthopoxviruses. The detection of the pathogens is possible without additional cultivation. In order to exclude false-negative PCR results the amplification reaction was established as a competitive PCR in which an internal control DNA is included as an amplification check. With regard to a possible automation the detection-systems were constituted in form of a 'PCR ELISA'. The colorimetric detection of the amplicons takes place thereby in streptavidin coated microtiter plates. For each system biotinylated capture probes for both the internal control DNA (ST) and the pathogen specific DNA (WT) have been developed. For the use of these proof systems in the clinical laboratory, the routine application of the tested decontamination procedure by means of UNG during the execution of the PCR ELISA is recommended. For all three systems a detection of the pathogens is possible within one working day. For the species specific detection of F. tularensis and the genus specific detection of Orthopoxviruses the PCR ELISA systems established here represent the first developments of this type with integrated amplification control and potential for quantification.