Triplex High-Resolution Melting Assay for the Simultaneous Assessment of IFNL3 rs12979860, ABCB11 rs2287622, and RNF7 rs16851720 Genotypes in Chronic Hepatitis C Patients

Abstract

Chronic hepatitis C (CHC) is a leading cause of liver disease. Despite the improved efficacy of new antivirals, their high costs preclude their adoption in resource-limited settings, where CHC prevalence is highest. We developed a triplex high-resolution melting assay for the simultaneous assessment of three genetic polymorphisms related to the response to treatment and development of advanced fibrosis in CHC: IFNL3 rs12979860, ABCB11 rs2287622, and RNF7 rs16851720. We validated the assay in clinical samples from 130 CHC patients treated with classic therapy. The assay showed excellent reproducibility and 100% accuracy, sensitivity, and specificity against the gold standard Sanger sequencing. When added to routine examination data, genotype information significantly improved their performance for prediction of advanced liver fibrosis and sustained virological response (P = 0.041 and P = 0.011, respectively). Correspondingly, the full models had area under the receiver operating characteristic curve values of 0.842 (95% CI, 0.773-0.911) and 0.921 (95% CI, 0.870-0.972) and integrated discrimination improvements of 7.5% (95% CI, 2.5%-12.5%; P = 0.003) and 11.5% (95% CI, 5.8%-17.2%; P < 0.001), respectively. This is the first report on a diagnostic test for simultaneous genotyping of IFNL3, ABCB11, and RNF7 in CHC patients. Reliable and inexpensive, the assay should provide useful information for the clinical management of CHC, like identification of patients at risk of rapid disease progression or with high chances of response to classic therapy.

HCV-Associated Nephropathies in the Era of Direct Acting Antiviral Agents

“…While the impact of the different HCV genotypes on renal outcomes still needs to be completely elucidated, careful renal function evaluation should be part of regular follow-up of individuals with HCV infection, especially if serum levels of HCV RNA are elevated and in case of infection with HCV genotypes 1 or 2. Moreover, in last decade several genome-wide association studies reported many host genetic factors that influence hepatic outcomes and treatment efficacy after HCV infection (21–23). A GWAS among patients with chronic HCV infection found a genome-wide significant association of rs9461776 ( HLA-DRB1/DQA1 ) with cryoglobulin-related vasculitis (24) The same study also identified single nucleotide polymorphisms (SNPs) near NOTCH4 and MHC class II that were strongly associated with this syndrome.…”