Useful molecular tools for facing next pandemic events: Effective sample preparation and improved RT-PCR for highly sensitive detection of SARS-CoV-2 in wastewater environment.

Viral pandemics can be inevitable in the next future. Considering SARS-CoV-2 pandemics as an example, there seems to be a need to develop a surveillance system able to monitor the presence of potential pathogenic agents. The sewage and wastewater environments demonstrated to be suitable targets for such kind of analysis. In addition, it is important to have reliable molecular diagnostic tools and also to develop a robust detection strategy. In this study, an effective sample preparation procedure was selected from four options and combined with a newly developed improved RT-PCR. First, a model viral system was constructed, containing a fragment of the SARS-CoV-2 gene encoding for the Spike protein. The encapsidated S RNA mimic (ESRM) was based on the plum pox virus (PPV) genome with the inserted targeted gene fragment. ESRM was used for seeding wastewater samples in order to evaluate the viral recovery of four different viral RNA concentration/extraction methods. The efficiency of individual approaches was assessed by the use of a quantitative reverse transcription PCR (qRT-PCR) and by a one-step single-tube nested quantitative reverse transcription PCR (OSN-qRT-PCR). For the detection of viruses in wastewater samples with low viral loads, OSN-qRT-PCR assay produced the most satisfactory results and the highest sensitivity.

Electrospun Poly(ethylene Terephthalate)/Silk Fibroin Composite for Filtration Application.

In this study, fibrous membranes from recycled-poly(ethylene terephthalate)/silk fibroin (r-PSF) were prepared by electrospinning for filtration applications. The effect of silk fibroin on morphology, fibers diameters, pores size, wettability, chemical structure, thermo-mechanical properties, filtration efficiency, filtration performance, and comfort properties such as air and water vapor permeability was investigated. The filtration efficiency (FE) and quality factor (Qf), which represents filtration performance, were calculated from penetration through the membranes using aerosol particles ranging from 120 nm to 2.46 µm. The fiber diameter influenced both FE and Qf. However, the basis weight of the membranes has an effect, especially on the FE. The prepared membranes were classified according to EN149, and the most effective was assigned to the class FFP1 and according to EN1822 to the class H13. The impact of silk fibroin on the air permeability was assessed. Furthermore, the antibacterial activity against bacteria S. aureus and E. coli and biocompatibility were evaluated. It is discussed that antibacterial activity depends not only on the type of used materials but also on fibrous membranes' surface wettability. In vitro biocompatibility of the selected samples was studied, and it was proven to be of the non-cytotoxic effect of the keratinocytes (HaCaT) after 48 h of incubation.