Rapid point-of-care HCV RNA quantification in capillary whole blood for diagnosing chronic HCV infection, monitoring treatment and detecting reinfection

BACKGROUND: Rapid point-of-care capillary hepatitis C virus (HCV) RNA quantification could remove barriers to chronic hepatitis C diagnosis and treatment. AIMS: To evaluate the diagnostic accuracy of rapid point-of-care HCV RNA quantification by Cepheid (R)'s GeneXpert (R) in 100 mu l capillary whole blood using our laboratory-based standard quantitative HCV polymerase chain-reaction (PCR) test (Roche Cobas (R) Ampliprep/Taqman) with 650 mu l venous EDTA plasma as the reference test. METHODS: In a prospective study conducted between November 2016 and May 2019 in the Infectious Diseases Outpatient Clinic of a Swiss tertiary care hospital, all adults with an indication for HCV RNA quantification (including HCV treatment monitoring) and written informed consent provided venous and capillary blood for parallel testing. Up to October 2018, we used the Xpert (R) HCV Viral Load (VL) test (105 min; developed for 1 ml plasma or serum), for which 1 ml Cepheid (R) buffer was added to 100 mu l finger-stick capillary whole blood (similar to 55% plasma). Thereafter, the Xpert (R) HCV Viral Load Finger-Stick (VL FS) test (60 min; specifically developed for 100 mu l capillary whole blood) was evaluated. RESULTS: (1) Xpert (R) HCV VL test. Among 194 paired samples from 88 patients, 99 (51.0%) were positive using Cobas (R) in venous plasma. Sensitivity and specificity of the Xpert (R) HCV VL test with 100 mu l capillary whole blood was 97.0% (96/99; 95% confidence interval [CI] 91.5-99.0%) and 94.7% (90/95; 95% CI 88.3-97.7%), respectively. The eight (4.1%) discordant results (three false negative, five false positive) were all under direct acting antiviral (DAA) treatment (week 1-4 or end of treatment), when HCV RNA was near the limit of quantification (highest HCV RNA value missed by Xpert (R) 68 IU/ml). Quantifiable results (n = 68) correlated well (R-2 = 0.9165) irrespective of genotype, sex and HIV status. On average, Xpert (R) HCV VL test results were 1.32 (+/- 0.34) log IU/ml lower, which corresponds to the similar to 18-fold smaller plasma volume used (similar to 55 vs 1000 mu l). (2) Xpert (R) HCV VL FS test: Among 33 paired samples from 23 patients, 15 (45.5%) were positive using Cobas (R) in venous plasma. Sensitivity and specificity of the Xpert (R) HCV VL FS test with 100 mu l capillary whole blood was 100% (15/15; 95% CI 79.6-100%) and 88.9% (16/18; 95% CI 67.2-96.9%), respectively. The two (6.1%) discordant results (both false positive) were under DAA treatment (week 3 and 4), when HCV RNA was near the limit of quantification. Quantifiable results (n = 14) correlated well (R-2 = 0.9899). On average, Xpert (R) HCV VL FS test results were 0.10 (+/- 0.17) log IU/ml lower. CONCLUSIONS: Point-of-care HCV RNA quantification in capillary whole blood is a convenient, rapid and reliable method to diagnose active HCV infection, monitor treatment response and detect reinfection. For patients with difficult venous access after long-term intravenous drug use, capillary testing removes a crucial barrier to HCV treatment and reinfection monitoring. Same-day results might improve linkage to care.