20.500.12556/DiRROS-19692 Enhanced detection of pathogenic enteric viruses in coastal marine environment by concentration using methacrylate monolithic chromatographic supports paired with quantitative PCR Currently, around 50% of the world's population lives in towns and cities within 100 km of the coast. Monitoring of viruses that are frequently present in contaminated coastal environments, such as rotavirus (RoV) and norovirus (NoV), which are also the major cause of human viral gastroenteritis, is essential to ensure the safe use of these water bodies. Since exposure to as few as 10–100 particles of RoV or NoV may induce gastrointestinal disease, there is a need to develop a rapid and sensitive diagnostic method for their detection in coastal water samples. In this study, we evaluate the application of methacrylate monolithic chromatographic columns, commercially available as convective interaction media (CIM®), to concentrate pathogenic enteric viruses from saline water samples prior to virus quantification by one-step reverse transcription quantitative PCR (RT-qPCR). Using RoV and NoV as model enteric viruses, we present our results on the most effective viral concentration conditions from saline water matrices using butyl (C4) hydrophobic interaction monolithic support (CIM® C4). C4 monolithic columns exhibit a good capacity to bind both RoV and NoV and both viruses can be eluted in a single step. Our protocol using a 1 ml C4 column enables processing of 400 ml saline water samples in less than 60 min and increases the sensitivity of RoV and NoV detection by approximately 50-fold and 10-fold respectively. The protocol was also scaled up using larger capacity 8 ml C4 columns to process 4000 ml of seawater samples with concentration factors of 300-fold for RoV and 40-fold for NoV, without any significant increase in processing time. Furthermore, C4 monolithic columns were adapted for field use in an on-site application of RoV concentration from seawater samples with performance equivalent to that of the reference laboratory setup. Overall, the results from successful deployment of CIM C4 columns for concentration of rotavirus and norovirus in seawater samples reiterate the utility of monolithic supports as efficient, scalable and modular preparative tools for processing environmental water samples to enhance viral detection using molecular methods. rotavirus norovirus seawater fecal contamination qPCR sewage true false true Angleški jezik Ni določen Neznano 2024-07-24 03:45:09 2024-07-25 17:52:17 2024-07-26 04:28:08 0000-00-00 00:00:00 2016 0 0 str. 405-414 Vol. 106 2016 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 2016-12-01 579 0043-1354 10.1016/j.watres.2016.10.020 4037967 RAZ_Balasubramanian_Mukundh_Narayanan_2016.pdf RAZ_Balasubramanian_Mukundh_Narayanan_2016.pdf 1 89071BF8CE518AF691F7AC6BEFD17394 935e22669f0a6f199d5952ee659df11ced902b47ed422387188149ecf68252b2 62d03988-4a8d-11ef-b487-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=27549 https://doi.org/10.1016/j.watres.2016.10.020 0 50d876bb-4a8d-11ef-b487-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=27548 Nacionalni inštitut za biologijo 0 0 0