Lopez E., Barthelemy M., Baronti Cécile, Masse S., Falchi A., Durbesson F., Vincentelli R., Lamballerie X. de, Charrel R., Coutard B. (2021). Endonuclease-based genotyping of the RBM as a method to track the emergence or evolution of SARS-CoV-2 variants. Iscience, 24 (11), p. 103329 [11 p.].
Titre du document
Endonuclease-based genotyping of the RBM as a method to track the emergence or evolution of SARS-CoV-2 variants
Lopez E., Barthelemy M., Baronti Cécile, Masse S., Falchi A., Durbesson F., Vincentelli R., Lamballerie X. de, Charrel R., Coutard B.
Source
Iscience, 2021,
24 (11), p. 103329 [11 p.]
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.
Plan de classement
Sciences fondamentales / Techniques d'analyse et de recherche [020]
;
Entomologie médicale / Parasitologie / Virologie [052]
