Development of forensic assay signatures for ebolaviruses

Area of Science:

• Virology
• Forensic Science
• Molecular Biology

Background:

• Ebolaviruses are a group of RNA viruses with five species, four of which are highly lethal to humans.
• Due to their high infectivity and lethality, ebolaviruses are classified as significant biothreat agents.
• Current detection assays lack forensic capabilities for differentiating ebolavirus species and variants.

Purpose of the Study:

• To develop novel forensic assays for the precise differentiation of ebolavirus species and variants.
• To establish high-resolution genotyping tools for ebolavirus forensic investigations.

Main Methods:

• Phylogenetic analyses were conducted to identify species-specific and clade-specific canonical single nucleotide polymorphisms (SNPs).
• TaqMan-MGB allelic discrimination assays were designed based on identified SNPs.
• Assays were screened against synthetic RNA templates and validated using ebolavirus genomic RNAs.

Main Results:

• A total of 45 validated assays demonstrated 100% coverage for ebolavirus species and variants, with resolution at the isolate level.
• Accurate forensic classification of four "unknown" ebolaviruses to the species and variant level was achieved.
• Resolution below the isolate level was not successfully attained with the developed assays.

Conclusions:

• The developed high-resolution forensic assays provide rapid and accurate genotyping of ebolaviruses.
• These assays are crucial tools for forensic investigations involving ebolavirus identification and characterization.
• The assays enhance biosecurity by enabling precise forensic analysis of potential ebolavirus threats.