Screening of an enterovirus specific RT-PCR ELISA method for the quantification of enterovirus genomes in human body fluids by means of a three-levelexperimental design

In order to obtain a detection limit as low as possible for a quantitative enterovirus specific RT-PCR ELISA assay, optimal reaction conditions, which give rise to the highest response, need to be determined. This was done by investigating the influence of 13 factors, selected from RT and PCR, in a multivariate approach by means of a well-balanced three-level screening des ign, derived from a three-level Plackett-Burman design. Optimal reaction co nditions could be determined by calculation and evaluation of the effects o f the different factors on the response. i.e. the measured absorbance of th e ELISA detection. The method will be used to study a possible longitudinal relationship between enteroviruses and the development of multiple scleros is and juvenile diabetes. (C) 2001 Elsevier Science B.V. All rights reserve d.