IN-HOUSE ASSAY SET UP FOR GENOTYPING AND RESISTANCE PROFILE OF HEPATITIS C (HCV) BY SEQUENCING THE NS5B (CENTRAL REGION) AND NS3 USING A SINGLE METHOD

Hepatitis C virus (HCV) infection remains a major public health concern affecting approximately 185 million people worldwide (Childs-Kean, L.M et al.). There is a high variability between circulating strains of hepatitis C virus. This variability determines and influences the response and duration of treatment (Petruzziello, A. et al.). Currently, there is limited data available on the HCV viral strains in Rwanda and there is no systematic laboratory genotyping and sequencing tools for optimal management of patients in the country. The purpose of this work is to develop and set up an in-house method within the Clinical Microbiology Laboratory in the CHU-Liege that allow simultaneous genotyping and Direct acting antivirals (DAAs) resistance profiling by direct sequencing the HCV NS3 and NS5B genes which is applicable to a wide range of genotypes. The developed tool will be subsequently transferred to Rwanda. This will aid in determining the HCV genotype among infected patients; ascertaining the epidemiological surveillance of circulating strains in Rwanda; and help in monitoring of patients under antiviral treatment in various health care facilities. The first objective was to validate the detection and identification of the common circulating HCV genotypes 1-6 using in-lab developed or literature cited primes. The second objective was to evaluate the sensitivity of the method and its capacity to monitor resistance of HCV to antiviral treatment especially to DAAs currently used in various clinical settings; and finally, to analyze clinical samples from patients routinely followed up in CHU of Liege.