Möglichkeiten und Grenzen beim Einsatz von Indikatorviren zur Überwachung virusbelasteter Oberflächengewässer

Nils Marten Hartmann, Hans-Christoph Selinka, Technische Universität Berlin, Technische Universität Berlin
2013
Human pathogenic viruses, released into the environment by insufficient or untreated sewage, may be a threat to public health. Current indicators for fecal pollution do not include viruses and their ability to survive outside their hosts and their properties differ from those of the viruses. Therefore, potential candidates, which could be used as viral indicators for sewage pollution are investigated around the world. Furthermore, reliable detection systems are needed, to ensure meaningful risk
more » ... ure meaningful risk assessments. For this purpose, different concentration methods were tested and compared with regard to recovery rates of viruses in native and spiked laboratory and environmental samples. Human pathogenic adenoviruses and noroviruses as well as enteric bacteriophages were used for spike experiments. All of these were previously regarded as potential indicator viruses. Results show, that high recoveries can be received even from samples with large volumes, using the tested monolithic affinity filtration system. However, in contrast to traditional methods, like glass wool filtration and ultracentrifugation, results varied and further evaluation is required. While adenoviruses and noroviruses were quantified by quantitative real-time polymerase chain reaction (qPCR), quantification of phages was performed by plaque assays. Detection methods may be subject to inhibitory substances within environmental samples. In particular, quantitative real-time polymerase chain reactions are at risk to fail, when inhibitors are present. Methods were investigated in order to eliminate relevant substances from environmental samples. By combining a nucleic acid extraction using the NucliSens extraction kit with MobiSpin column purification, quantification was possible in laboratory samples containing up to 400 ng/μl of humic acids. Without further treatment reliable quantification was not possible above 4 ng/μl. Moreover, application of MobiSpin columns improved quantification limits in inhibited environmental samples. The quality of qPCR resu [...]
doi:10.14279/depositonce-3722 fatcat:ctqwpqni2rhx5on7v3y3j5vdge