@article{fdi:010078936,
  title = {{L}yophilized matrix containing ready-to-use primers and probe solution for standardization of real-time {PCR} and {RT}-q{PCR} diagnostics in virology},
  author = {{T}hirion, {L}aurence and {D}ubot {P}{\'e}r{\`e}s, {A}udrey and {P}ezzi, {L}. and {C}orcostegui, {I}. and {T}ouinssi, {M}. and {L}amballerie, {X}. de and {C}harrel, {R}. {N}.},
  editor = {},
  language = {{ENG}},
  abstract = {{R}eal-time molecular techniques have become the reference methods for direct diagnosis of pathogens. {T}he reduction of steps is a key factor in order to decrease the risk of human errors resulting in invalid series and delayed results. {W}e describe here a process of preparation of oligonucleotide primers and hydrolysis probe in a single tube at predefined optimized concentrations that are stabilized via lyophilization ({L}yoph-{P}&{P}). {L}yoph-{P}&{P} was compared versus the classic protocol using extemporaneously prepared liquid reagents using (i) sensitivity study, (ii) long-term stability at 4 degrees {C}, and (iii) long-term stability at 37 degrees {C} mimicking transportation without cold chain. {T}wo previously published molecular assays were selected for this study. {T}hey target two emerging viruses that are listed on the blueprint of the {WHO} as to be considered for preparedness and response actions: chikungunya virus ({CHIKV}) and {R}ift {V}alley fever phlebovirus ({RVFV}). {R}esults of our study demonstrate that (i) {L}yoph-{P}&{P} is stable for at least 4 days at 37 degrees {C} supporting shipping without the need of cold chain, (ii) {L}yoph-{P}&{P} rehydrated solution is stable at +4 degrees {C} for at least two weeks, (iii) sensitivity observed with {L}yoph-{P}&{P} is at least equal to, often better than, that observed with liquid formulation, (iv) validation of results observed with low-copy specimens is rendered easier by higher fluorescence level. {I}n conclusion, {L}yoph-{P}&{P} holds several advantages over extemporaneously preparer liquid formulation that merit to be considered when a novel real-time molecular assay is implemented in a laboratory in charge of routine diagnostic activity.},
  keywords = {freeze-drying ; lyophilization ; {PCR} ; diagnosis ; virus ; pathogen ; {T}aq{M}an ; emerging ; epidemic},
  booktitle = {},
  journal = {{V}iruses},
  volume = {12},
  numero = {2},
  pages = {art. 159 [13 ]},
  year = {2020},
  DOI = {10.3390/v12020159},
  URL = {https://www.documentation.ird.fr/hor/fdi:010078936},
}