Reducing the incidence of hepatocellular carcinoma due to hepatitis C virus infection by detecting resistanceassociated substitutions in hepatitis C virus NS5A nonstructural protein

Objective: Hepatocellular carcinoma (HCC) is one of the leadingcauses of liver cancer worldwide. In Malaysia, liver cancer is oneof the main cancers detected in the population with an annualmortality rate of 6.1% in 2013. Hepatitis C virus (HCV) infectionis one of the causes attributed to the incidence of HCC. HCVinfection remains a major concern in Malaysia as many peopleare unaware of their infection status. With the accessibility andeffectiveness of Direct Acting Agents (DAAs), more people canbe screened and diagnosed with hepatitis C. With the access andaffordability of sofosbuvir and daclatasvir in the country, highcure rate among treated HCV infected patients can be achieved;therefore, this will reduce the occurrence of HCC. However,there is a need to detect the presence of resistance-associatedsubstitutions (RASs) to better treat HCV-associated HCC. Thisstudy aims to develop an in-house HCV drug resistance assayand detect RASs that confer resistance to NS5A inhibitors amongnewly diagnosed HCV infected patients. Methods: Frozen plasma samples used in the study wereobtained from hospitals in Malaysia as part of routinediagnostic services. An extensive literature search wasconducted to choose the appropriate HCV drug resistancegenotyping assay and the two sets of primers to be adoptedand used for detecting RASs for NS5A inhibitors among HCVgenotype 3 patients in the country. The DNA sequences ofHCV strains covering the NS5A region were submitted to thegeno2pheno (HCV) resistance database created by Max PlanckInstitute (MPI) Informatics to yield drug-related mutations. Results: The assay was optimized, and all samples weresuccessfully amplified by using set 2 of the chosen primers.Amplicons that were about 1.4 kb in size were producedfrom the samples and positively sequenced. Good qualitychromatograms were produced for all samples and contigmaps were generated. Drug resistance results against NS5Ainhibitors such as daclatasvir, elbasvir, ledipasvir, pibrentasvir,and velpatasvir were generated. Conclusions: With the availability of the optimized in-houseHCV drug resistance assay in Malaysia, HCV-infected patientscan be treated and managed effectively, and therefore, a highcure rate can be obtained. DOI: 10.20892/j.issn.2095-3941.2018.S054