Kham-Kjing N., Ngo-Giang-Huong Nicole, Tragoolpua K., Khamduang W., Hongjaisee S. (2022). Highly specific and rapid detection of hepatitis c virus using RT-LAMP-coupled CRISPR-Cas12 assay. Diagnostics, 12 (7), p. 1524 [11 p.].
Titre du document
Highly specific and rapid detection of hepatitis c virus using RT-LAMP-coupled CRISPR-Cas12 assay
Kham-Kjing N., Ngo-Giang-Huong Nicole, Tragoolpua K., Khamduang W., Hongjaisee S.
Source
Diagnostics, 2022,
12 (7), p. 1524 [11 p.]
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.
