Development of Forensic Assay Signatures for Ebolaviruses |
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| Authors: | Jian Song PhD Norman Doggett PhD Melinda Wren MS Tom Burr PhD P W Fenimore PhD Eneida L Hatcher MS William J Bruno PhD Po‐E Li MS Chris Stubben PhD Murray Wolinsky PhD |
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| Institution: | 1. Bioenergy and Biome Sciences (B‐11), Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM;2. Biosecurity and Public Health (B‐10), Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM;3. Statistical Sciences (CCS‐6), Computer, Computational, and Statistical Sciences Division, Los Alamos National Laboratory, Los Alamos, NM;4. Theoretical Biology and Biophysics (T‐6), Theoretical Division, Los Alamos National Laboratory, Los Alamos, NM;5. Department of Microbiology, University of Alabama Birmingham, Birmingham, AL |
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| 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. |
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| Keywords: | forensic science DNA typing hemorrhagic fever Ebolavirus Ebola filovirus filoviridae canonical SNPs SNP genotyping TaqMan‐MGB allelic discrimination assay |
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