@article{fdi:010077829,
  title = {{D}evelopment and evaluation of a duo {Z}aire ebolavirus {R}eal-{T}ime {RT}-{PCR} assay targeting two regions within the genome},
  author = {{T}hirion, {L}aurence and {C}harrel, {R}. {N}. and {B}oehmann, {Y}. and {C}orcostegui, {I}. and {R}aoul, {H}. and {L}amballerie, {X}. de},
  editor = {},
  language = {{ENG}},
  abstract = {{P}reparedness and response actions to mitigate {E}bola virus disease ({EVD}) outbreaks rely on rapid diagnosis to be implemented locally to sort suspect patients attending health centers. {O}ur aim was (i) to develop and evaluate an {RT}-q{PCR} assay combining primers and probes derived from two reference assays targeting different genomic regions; (ii) to study whether sensitivity and specificity of this dual-target assay were at least equal or better to the parental assays; (iii) to implement this dual-target assay onto the {C}epheid {G}ene{X}pert open cartridge as a proof of principle for technological transfer aiming at bedsite testing locally. {T}o do so, three home-made published {RT}-q{PCR} assays were selected to be compared with the {R}eal{S}tar(({R})) {F}ilovirus {S}creen {RT}-{PCR} kit 1.0 ({A}ltona {D}iagnostics, {H}amburg, {G}ermany), a technique that was largely deployed during the 2014-2015 {W}est {A}frican {EVD} outbreak. {P}rimers and probes sequences of the custom-made assays were analyzed in silico against a multiple sequence alignment, including >250 complete sequences corresponding to strains that have caused {EVD} epidemics in the past. {G}enomic {RNA} purified from the {M}ekambo strain of {Z}aire ebolavirus ({EBOV}) was used to study the sensitivity of the five methods. {B}ased on these results, two in-house methods were selected and adapted to design the dual-target assay, which performances were compared to those of the parental assays using a synthetic {RNA} control. {T}he dual-target assay showed better sensitivity and limit of detection ({L}o{D}(95) at 0.4 copies/mu {L}) than the parental methods (1.7 and 2.2 copies/mu {L}). {U}ltimately, the dual-target assay was transferred onto the {G}ene{X}pert {F}lex-03 open cartridge, demonstrating a {L}o{D}(95) at 0.75 copies/mu {L}. {T}ogether these results indicate that {EBOV} dual-target assay has the potential to be used during {EVD} outbreak in the laboratory having performed molecular testing during the recent outbreaks.},
  keywords = {filoviridae ; ebola virus disease ; diagnostics ; emerging ; viral ; hemorrhagic fever ; preparedness ; response ; {ZAIRE}},
  booktitle = {},
  journal = {{M}icroorganisms},
  volume = {7},
  numero = {12},
  pages = {art. 652 [15 ]},
  year = {2019},
  DOI = {10.3390/microorganisms7120652},
  URL = {https://www.documentation.ird.fr/hor/fdi:010077829},
}