%0 Journal Article
%9 ACL : Articles dans des revues avec comité de lecture répertoriées par l'AERES
%A Lopez, E.
%A Barthelemy, M.
%A Baronti, Cécile
%A Masse, S.
%A Falchi, A.
%A Durbesson, F.
%A Vincentelli, R.
%A Lamballerie, X. de
%A Charrel, R.
%A Coutard, B.
%T Endonuclease-based genotyping of the RBM as a method to track the emergence or evolution of SARS-CoV-2 variants
%D 2021
%L fdi:010083867
%G ENG
%J Iscience
%M ISI:000730171000001
%N 11
%P 103329 [11 ]
%R 10.1016/j.isci.2021.103329
%U https://www.documentation.ird.fr/hor/fdi:010083867
%> https://horizon.documentation.ird.fr/exl-doc/pleins_textes/2022-01/010083867.pdf
%V 24
%W Horizon (IRD)
%X Since the beginning of the COVID-19 pandemics, variants have emerged. Some of them display increased transmissibility and/or resistance to immune response. Most of the mutations involved in the functional adaptation are found in the receptor-binding motif (RBM), dose to the interface with the receptor ACE2. We thus developed a fast molecular assay to detect mutations in the RBM coding sequence. After amplification, the amplicon is heat-denatured and hybridized with an amplicon of reference. The presence of a mutation can be detected using a mismatch-specific endonuclease and the cleavage pattern is analyzed by capillary electrophoresis. The method was validated on RNA of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants produced in vitro before being implemented for clinical samples. The assay showed 97.8% sensitivity and 97.8% specificity. The procedure can be set up for high-throughput identification of the presence of mutations and serve as a first-line screening to select the samples for full genome sequencing.
%$ 052 ; 020