Development and evaluation of a method for concentration and detection of salmonid alphavirus from seawater
| Autor: | Atle Lillehaug, Lisa-Victoria Bernhardt, Mette Myrmel, Lars Qviller, Simon Chioma Weli |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Salmo salar 030106 microbiology Alphavirus Biology Real-Time Polymerase Chain Reaction Virus law.invention Fish Diseases 03 medical and health sciences Hydrophilic membrane law Virology Lysis buffer Animals Seawater Filtration Chromatography Alphavirus Infections Elution Fish health biology.organism_classification 030104 developmental biology Salmonidae |
| Zdroj: | Journal of Virological Methods |
| Popis: | Waterborne viral infections represent a major threat to fish health. For many viruses, understanding the interplay between pathogens, host and environment presents a major hurdle for transmission. Salmonid alphavirus (SAV) can infect and cause pancreas disease (PD) in farmed salmonids in seawater. During infection, SAV is excreted from infected fish to the seawater. We evaluated two types of filters and four different eluents, for concentration of SAV3. One L of seawater was spiked with SAV3, followed by filtration and virus elution from membrane filters. For the negatively charged MF hydrophilic membrane filter (MF-) combined with NucliSENS® lysis buffer the SAV3 recovery was 39.5 ± 1.8 % by RT-ddPCR and 25.9 ± 5.7 % by RT-qPCR. The recovery using the positively charged 1 MDS Zeta Plus® Virosorb® membrane filter (MD+), combined with NucliSENS® lysis buffer was 19.0 ± 0.1 % by RT-ddPCR and 13.3 ± 3.8 % by RT-qPCR. The limits of quantification (LOQ) and detection (LOD) were estimated to be 5.18 × 103 and 2.0 × 102 SAV3 copies/L of natural seawater, by RT-ddPCR. SAV3 recovery from small volumes of seawater, and the requirement for standard laboratory equipment, suggest the MF-filter combined with NucliSENS® lysis buffer would be a candidate for further validation in experimental trials. |
| Databáze: | OpenAIRE |
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