Hi. I am collected blood from patients with a BSL-4 virus onto filter paper, i.e dried blood spots. We are running some validation experiments to see whether the drying process inactive the virus. So we need to elute the blood/virus from the filter paper and then culture the virus. The methods of elution that I can find for DBS seem likely to inactive the virus itself, and therefore are not suitable since this would prevent us from culturing the virus, and seeing whether drying itself inactive. We can't use sonication because of H&S issues inside the BSL4 lab.
Does anyone know of any elution methods that would not generally inactivate viruses?
Thanks
Elution of Dried Blood Spots
1. Punch out one spot with a single-use 6 mm device from each blood-soaked circle of the Grade 903 filter card (Figure 1F). Transfer all punched dried blood spots from a single patient to one well of the 12-well plate.
2. Fill the well with phosphate-buffered saline containing 0.05% Tween 20 and 0.08% sodium azide (Figure 1G). Adapt the volume of added buffer to the minimal respective requirements of the assay used for subsequent analysis of dried blood spots eluates.
3. Repeat these steps to obtain a second series of dried blood spot eluates in order to perform molecular analyses.
4. Put the cell culture plate on a laboratory shaker and let the punched dried blood spots gently elute for a minimum of 4 hr or, preferably, O/N (Figure 1H).
5. The next day, the spots are almost free from blood and hemolytic supernatants have formed (Figure 1I). Transfer these eluates to microcentrifuge tubes. Then, subject them to centrifugation for 2 min at 10,500 x g (Figure 1J) to free the supernatants from any debris that had formed during elution (Figure 1K).
Thanks, I think the issue here is that 0.05% Tween 20 and 0.08% sodium azide and leaving for 4 hour-overnight may itself potentially inactive the virus. We want to avoid any elution methods that would inactive the virus, so that we test whether the drying alone inactivates the virus. Thanks
Hello, see attached fill paper which contain same info that you described.
Good luck!
- Badges
- Science topic
- Similar topics
- Biological Science
- Microbiology
- Virology
- Virus