Species-specific identification of variola, monkeypox, cowpox, and vaccinia viruses by multiplex real-time PCR assay

The Microbial Detection Array for Detection of Emerging Viruses in Clinical Samples - A Useful Panmicrobial Diagnostic Tool

“…To confirm presence of virus in the samples and quantify the virus before and after WTA, we performed virus-specific real-time PCR. We used in-house assays for DENV, WNV, orthopoxvirus, parapoxvirus, Usutu virus, Hantaan virus, Toscana virus, BK virus (BKV) and rotavirus A; and previously published assays for JC virus (JCV) [30], cowpox and monkeypox viruses [31], Chikungunya virus [32], Eastern equine encephalitis virus (EEEV) [33], JEV [34], TBEV [35], YFV [36], Lassa virus and CCHFV [37], Dobrava-Belgrade virus (DOBV) [38], Puumala virus [39], Rift Valley fever virus (RVFV) [36] and Marburg virus [40]. PCR was performed using an Mx3005P (Stratagene) thermal cycler.…”

Orthopoxviruses—Plagues of Mankind, Strategists in Immune Evasion, Teachers in Vaccination

“…Since 2001, real-time PCRs have been developed (e.g. Czerny et al 2001;Kulesh et al 2004;Balamurugan et al 2009;Gavrilova et al 2010;Li et al 2010;Shchelkunov et al 2011;Venkatesan et al 2012a). A DNA oligonucleotide microarray with the TNF receptor gene crmB (Lapa et al 2002) and loopmediated isothermal amplification (LAMP) assays are alternative rapid methods for OPXV detection or differentiation (Iizuka et al 2009;Venkatesan et al 2012b).…”

Comparison of nucleic acid extraction platforms for detection of select biothreat agents for use in clinical resource limited settings

“…This is especially true for the potential of an intentional release of a biological agent. In this context, real-time PCR is currently implemented for the highly specific identification of pathogens in both clinical and environmental settings (Belgrader et al, 1998;Christensen et al, 2006;Higgins et al, 1998;Makino et al, 2001;Ryu et al, 2003;Shchelkunov et al, 2011;Skottman et al, 2007;Tomaso et al, 2008).…”