![\"Ebola](\"http:\/\/admin.acceleratingscience.com\/wp-content\/uploads\/2014\/10\/ebola_-_more_than_one.jpg\")
Filovirus hemorrhagic fever features\u00a0a rapid onset, high level of infectiousness, and staggering mortality rates of up to 90%.\u00a0Because\u00a0no clinically licensed therapeutic treatments or vaccines exist at this point in time, filovirus outbreaks\u2014including the\u00a0Ebola and Marburg viruses\u2014are a significant concern for both public\u00a0health and biodefense. For these reasons, the need for reliable, sensitive diagnostics is acute.<\/p>\n
Recently, Kamata et al., working out of the U.S. Army Medical Research Institute for Infectious Diseases, developed a protein microarray-based assay capable of detecting all members of the Filoviridae family: the Marburg virus (Marburg marburgvirus<\/em>) and the Ebola viruses (Sudan ebolavirus, Zaire ebolavirus, Reston ebolavirus, Bundibugyo ebolavirus <\/em>and Ta\u00ef Forest ebolavirus<\/em>).1<\/span><\/sup><\/p>\n To compose the assay, the research team took full-length genes for nucleoprotein (NP), virion protein 40 (VP40) and the glycoprotein mucin-like domain fragment (GP-mucin) for each virus species and cloned these into a pENTR\/TEV\/D-TOPO vector (Life Technologies, a\u00a0Thermo Fisher Scientific brand). Then they used LR Clonase II (Life Technologies) to shuttle entry vector clones into destination His-tagged expression vectors. The team\u00a0expressed and spotted a total of 34 purified recombinant proteins before microarray processing.<\/p>\n The team used anti-His antibody and a panel of purified filovirus antibodies to validate the assay, confirming that the microarray performed as designed. They determined that all His-tagged proteins were successfully spotted. Most of the antibodies bound with their targets with a high degree of specificity (anti-Marburg-VP40, anti-Marburg-GP, anti-Sudan-VP40, anti-Sudan-GP, anti-Zaire-VP40, anti-Zaire-NP, and anti-Zaire-GP), while some displayed minor cross-reactivity.<\/p>\n The researchers also evaluated the assay using serum samples from rhesus macaques vaccinated with trivalent virus-like particles (VLPs) prior to challenge with Marburg marburgvirus<\/em> (MARV) or Zaire ebolavirus<\/em> (ZEBOV). They collected three samples from each primate: naive, post-immunization, and post-challenge. The team applied the serum samples to their microarray, detecting bound IgG using fluorescently labeled antibodies.<\/p>\n