title: Development of forensic assay signatures for ebolaviruses.
abstract: Ebolaviruses are a diverse group of RNA viruses comprising five different species, four of which cause fatal hemorrhagic fever in humans. Because of their high infectivity and lethality, ebolaviruses are considered major biothreat agents. Although detection assays exist, no forensic assays are currently available. Here, we report the development of forensic assays that differentiate ebolaviruses. We performed phylogenetic analyses and identified canonical SNPs for all species, major clades and isolates. TaqMan-MGB allelic discrimination assays based on these SNPs were designed, screened against synthetic RNA templates, and validated against ebolavirus genomic RNAs. A total of 45 assays were validated to provide 100% coverage of the species and variants with additional resolution at the isolate level. These assays enabled accurate forensic analysis on 4 "unknown" ebolaviruses. Unknowns were correctly classified to species and variant. A goal of providing resolution below the isolate level was not successful. These high-resolution forensic assays allow rapid and accurate genotyping of ebolaviruses for forensic investigations.
authors: Bioenergy and Biome Sciences (B-11), Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM, 87545.
authors: Jian
authors: J
authors: Song
authors: Norman
authors: N
authors: Doggett
authors: Melinda
authors: M
authors: Wren
authors: Tom
authors: T
authors: Burr
authors: P W
authors: PW
authors: Fenimore
authors: Eneida L
authors: EL
authors: Hatcher
authors: William J
authors: WJ
authors: Bruno
authors: Po-E
authors: PE
authors: Li
authors: Chris
authors: C
authors: Stubben
authors: Murray
authors: M
authors: Wolinsky
mesh_terms: D000483:Alleles; D029043:Ebolavirus / Q000235:genetics*; D053803:Forensic Genetics; D016679:Genome, Viral; D010802:Phylogeny; D020641:Polymorphism, Single Nucleotide; D012367:RNA, Viral / Q000032:analysis; D060888:Real-Time Polymerase Chain Reaction; D017421:Sequence Analysis
chemical_list: D012367:RNA, Viral
keywords: DNA typing; Ebola; Ebolavirus; SNP genotyping; TaqMan-MGB allelic discrimination assay; canonical SNPs; filoviridae; filovirus; forensic science; hemorrhagic fever