%0 Journal Article
%9 ACL : Articles dans des revues avec comité de lecture répertoriées par l'AERES
%A Kham-Kjing, N.
%A Ngo-Giang-Huong, Nicole
%A Tragoolpua, K.
%A Khamduang, W.
%A Hongjaisee, S.
%T Highly specific and rapid detection of hepatitis c virus using RT-LAMP-coupled CRISPR-Cas12 assay
%D 2022
%L PAR00024851
%G ENG
%J Diagnostics
%K hepatitis C virus ; HCV RNA ; chronic hepatitis C ; point-of-care testing ; RT-LAMP ; CRISPR-Cas12 ; lateral flow-based assay ; fluorescence-based ; assay
%M ISI:000832298100001
%N 7
%P 1524 [11 ]
%R 10.3390/diagnostics12071524
%U https://www.documentation.ird.fr/hor/PAR00024851
%> https://horizon.documentation.ird.fr/exl-doc/pleins_textes/2022-09/010085900.pdf
%V 12
%W Horizon (IRD)
%X Hepatitis C virus (HCV) infection can be cured with pan-genotypic direct-acting antiviral agents. However, identifying individuals with current hepatitis C remains a major challenge, especially in resource-limited settings where access to or availability of molecular tests is still limited. The goal of this study was to develop and validate a molecular assay for the rapid detection of HCV RNA in resource-limited settings. It is based on a combination of reverse transcription loop-mediated isothermal amplification (RT-LAMP) with the clustered regularly interspaced short palindromic repeats-CRISPR-associated protein 12a (CRISPR-Cas12a) cleavage assay that allows the recognition of specific HCV nucleic acid sequences. Amplified products after the cleavage reactions can be visualized on lateral flow strips or measured with a fluorescence detector. When tested on clinical samples from individuals infected with HCV, HIV, or HBV, or from healthy donors, the RT-LAMP-coupled CRISPR-Cas12 assay yielded 96% sensitivity, 100% specificity, and 97% agreement as compared to the reference method (Roche COBAS AmpliPrep/COBAS TaqMan HCV Test). This assay could detect HCV RNA concentrations as low as 10 ng/mu L (an estimated 2.38 Log(10) IU/mL). Therefore, this sensitive and specific assay may represent an affordable and reliable point-of-care test for the identification of individuals with active hepatitis C in low-resource settings.
%$ 052 ; 050