Purification of Bacterial RNA from Infected Macrophages

Evaluation of a Bioaerosol Sampler for Indoor Environmental Surveillance of Severe Acute Respiratory Syndrome Coronavirus 2

“…To this end, this study evaluated the ability of multiple media to capture, stabilize, and integrate into a standard SARS-CoV-2 molecular diagnostic workflow 46 . The media tested had previously demonstrated promise for the collection of viral RNA [24][25][26][27][28][29][30][31][32][33][34][35][36][37] , either as a media typically associated with prolonged stability of the collected nucleic acids, previous use in aerosolized virus collection, or use in clinical specimen collection. In bench-scale trials, there was a significant link between media that were capable of reliably releasing stored viral supernatants (ACM, flocked swab, cotton swab) and higher concordance to viral supernatant controls.…”

“…Bench Series 1: Capture Media Testing Five unique media types including flocked swabs (Typenex, Catalog #SW0202) [25][26][27][28][29] , cotton swabs (Puritan, Catalog #25-8061WC) 25,26,28,29 , glass fiber filters (Millipore, Catalog #HAWP04700) [30][31][32][33] , FTA cards (Whatman, Catalog #29277432) [34][35][36][37] , and AerosolSense Capture Media (ACM) that have demonstrated previous success in nucleic acid isolation were tested within an aerosol sampling platform 24 (Thermo Fisher Scientific, Catalog #2900-AA). SARS-CoV-2 deposited by the Centers for Disease Control and Prevention (CDC) and obtained through BEI Resources, NIAID, NIH (SARS-Related Coronavirus 2, Isolate USA-WA1/2020, NR-52281) were cultured using Vero E6 cells (ATCC CRL-1586) for three to four days in Dulbecco's Minimum Essential Medium (DMEM, ATCC, Catalog #30-2002) at 5% CO2.…”

Untitled

“…To this end, this study evaluated the ability of multiple types of media to capture, stabilize, and integrate into a standard SARS-CoV-2 molecular diagnostic workflow [48]. The media tested had previously demonstrated promise for the collection of viral RNA [24][25][26][27][28][29][30][31][32][33][34][35][36][37], either as a media typically associated with prolonged stability of the collected nucleic acids, previous use in aerosolized virus collection, or use in clinical specimen collection. In bench-scale trials, there was a significant link between media that were capable of reliably releasing stored viral supernatants (ACM, flocked swab, cotton swab) and higher concordance to viral supernatant controls.…”

“…Five unique media types including flocked swabs (Typenex, Catalog #SW0202) [25][26][27][28][29], cotton swabs (Puritan, Catalog #25-8061WC) [25,26,28,29], glass fiber filters (Millipore, Catalog #HAWP04700) [30][31][32][33], FTA cards (Whatman, Catalog #29277432) [34][35][36][37], and Aerosol-Sense Capture Media (ACM) that have demonstrated previous success in nucleic acid isolation were tested within an aerosol sampling platform [24] (Thermo Fisher Scientific, Catalog #2900-AA). SARS-CoV-2 deposited by the Centers for Disease Control and Prevention (CDC) and obtained through BEI Resources, NIAID, NIH (SARS-Related Coronavirus 2, Isolate USA-WA1/2020, NR-52281) were cultured using Vero E6 cells (ATCC CRL-1586) for three to four days in Dulbecco's Minimum Essential Medium (DMEM, ATCC, Catalog #30-2002) at 5% CO 2 .…”