Hv durapore membrane stericup sterile vacuum filtration system

The Romero strain of JUNV (GenBank accession nos. AY619640 and AY619641) was obtained from Dr. Thomas G. Ksiazek (Centers for Disease Control and Prevention, Atlanta, GA). The virus was isolated from a patient and passaged twice in fetal rhesus lung cells and once in Vero cells [28 (link)]. Laboratory virus stock was obtained by amplification in Vero E6 cells. Cell debris in supernatants were filtered out through 0.45 μm HV Durapore Membrane Stericup sterile vacuum filtration system (Millipore Corporation, Billerica, MA). Cleared supernatants were concentrated through 30 min centrifugation at 3220 × g using Amicon Ultra-15 Centrifugal Filter Unit PLHK Ultracel-PL Membrane, 100 kDa (Millipore Corporation, Billerica, MA). All work with JUNV was performed at the University of Texas Medical Branch BSL-4 facilities (Robert E. Shope Laboratory) in accordance with institutional health and safety guidelines and federal regulations [29 ].

Romero (GenBank accession nos. AY619640 and AY619641) [44] (link) and Candid#1 (GenBank accession no. U70801) strains of JUNV were obtained from Dr. Thomas G. Ksiazek (Centers for Disease Control and Prevention, Atlanta, GA) and Dr. Robert Tesh (The World Reference Center for Emerging Viruses and Arboviruses (WRCEVA), University of Texas Medical Branch, Galveston, TX), respectively. Virus stocks were prepared using Vero cells. Cell debris in supernatants were filtered out through 0.45 µm HV Durapore Membrane Stericup sterile vacuum filtration system (Millipore Corporation, Billerica, MA). Cleared supernatants were concentrated through 30 min centrifugation at 3220×g using Amicon Ultra-15 Centrifugal Filter Unit PLHK Ultracel-PL Membrane, 100 kDa (Millipore Corporation, Billerica, MA). All work with JUNV Romero was performed at the University of Texas Medical Branch BSL-4 facilities (Robert E. Shope Laboratory or the Galveston National Laboratory) in accordance with institutional health and safety guidelines and federal regulations [45] .