Development of a detection system for waterborne pathogens (EDIT)

Molecular biological live/dead differentiation

In the EDIT project, an innovative monitoring system was developed with several project partners to swiftly detect viruses and bacteria in drinking water and raw water.

Besides ensuring the high quality of drinking water, drinking water suppliers are extremely concerned with the qualitative evaluation of the used raw water. In practice, the microbiological water quality is assessed using indicator bacteria. Owing to the extremely high expense, it is hardly possible to fully monitor all known microorganisms.

Indicator bacteria are detected in culture techniques through their growth in culture media. This evidence needs 18 hours for quickly proliferating bacteria such as E. coli and can take up to two weeks, especially for viruses.

Rapid detection of all hygienically relevant organisms is possible using molecular biological detection methods. These have long been established in medical diagnostics, however their low sensitivity makes them unsuitable for direct application in the drinking water domain.

In the BMBF project EDIT a multi-step procedure was developed and tested for the rapid molecular biological detection of viruses and bacteria.

In the project, TZW was responsible for developing the live/dead differentiation as well as for validating the detection system. If a bacterium or virus is inactivated, it can still be detected using molecular biological methods. However, for hygienic evaluations and risk assessments, the only organisms that matter are those that can trigger an infection.

Dead bacteria and inactivated viruses could be excluded from molecular biological detection in laboratory experiments by using fluorescent dyes. Within the project, various disinfection methods such as ozone, chlorine and UV radiation were also investigated and tested using the method.

Publications:

Elsäßer D., Ho J., Niessner R., Tiehm A., Seidel M.: Heterogeneous asymmetric recombinase polymerase amplification (haRPA) for rapid hygiene control of large-volume water samples. Analytical Biochemistry 546: 58-64 (2018) DOI 10.1016/j.ab.2018.01.032

Ho J., Seidel M., Niessner R., Eggert J., Tiehm A.: Long amplicon (LA)-qPCR for the discrimination of infectious and noninfectious phiX174 bacteriophages after UV inactivation. Water Research 103: 141-148 (2016) DOI 10.1016/j.watres.2016.07.032

Karthe D., Behrmann O., Blättel V., Elsässer D., Heese C., Hügle M., Hufert F., Kunze A., Niessner R., Ho J., Scharow B., Spoo M.,Tiehm A., Urban G., Vosseler S., Westerhoff T., Dame G., Seidel M.: Modular development of an inline monitoring system for waterborne pathogens in raw and drinking water. Environmental Earth Sciences 75: 1481 (2016) DOI 10.1007/s12665-016-6287-9

Lengger S., Otto J., Elsässer D., Schneider O., Tiehm A., Fleischer J., Niessner R., Seidel M.: Oligonucleotide microarray chip for the quantification of MS2, ?X174 and adenoviruses on the multiplex analysis platform MCR3. Analytical and bioanalytical chemistry, 406(14), 3323-3334 (2014) Doi: 10.1007/s00216-014-7641-y

Ho, J.: Molekularbiologische Lebend/tot-Unterscheidung [Molecular biological live/dead differentiation]. Veröffentlichungen aus dem Technologiezentrum Wasser [Publications from TZW], ISSN 1434-5765, TZW Volume 81 (2017)

Volumes from the TZW publication series can be purchased here.

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