Skip to main navigation menu Skip to main content Skip to site footer

Original article

Vol. 149 No. 4142 (2019)

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

  • Andrea Bregenzer
  • Nicole Warmann
  • Cornelia Ottiger
  • Christoph Andreas Fux
DOI
https://doi.org/10.4414/smw.2019.20137
Cite this as:
Swiss Med Wkly. 2019;149:w20137
Published
10.10.2019

Summary

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®’s GeneXpert® in 100 µl capillary whole blood using our laboratory-based standard quantitative HCV polymerase chain-reaction (PCR) test (Roche Cobas® Ampliprep/Taqman) with 650 µl 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® HCV Viral Load (VL) test (105 min; developed for 1 ml plasma or serum), for which 1 ml Cepheid® buffer was added to 100 µl finger-stick capillary whole blood (~55% plasma). Thereafter, the Xpert® HCV Viral Load Finger-Stick (VL FS) test (60 min; specifically developed for 100 µl capillary whole blood) was evaluated.

RESULTS

(1) Xpert® HCV VL test. Among 194 paired samples from 88 patients, 99 (51.0%) were positive using Cobas® in venous plasma. Sensitivity and specificity of the Xpert® HCV VL test with 100µl 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® 68 IU/ml). Quantifiable results (n = 68) correlated well (R2 = 0.9165) irrespective of genotype, sex and HIV status. On average, Xpert® HCV VL test results were 1.32 (±0.34) log IU/ml lower, which corresponds to the ~18-fold smaller plasma volume used (~55 vs 1000µl).

(2) Xpert® HCV VL FS test: Among 33 paired samples from 23 patients, 15 (45.5%) were positive using Cobas® in venous plasma. Sensitivity and specificity of the Xpert® HCV VL FS test with 100 µl 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 (R2 = 0.9899). On average, Xpert® 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.

References

  1. WHO. 05/2016, Combating hepatitis B and C to reach elimination by 2030 - Advocacy brief. http://apps.who.int/iris/bitstream/10665/206453/1/WHO_HIV_2016.04_eng.pdf (Accessed: 16 Jan. 2018)
  2. Chahine EB, Sucher AJ, Hemstreet BA. Sofosbuvir/Velpatasvir: The First Pangenotypic Direct-Acting Antiviral Combination for Hepatitis C. Ann Pharmacother. 2017;51(1):44–53. doi:.https://doi.org/10.1177/1060028016668897
  3. Puoti M, Foster GR, Wang S, Mutimer D, Gane E, Moreno C, et al. High SVR12 with 8-week and 12-week glecaprevir/pibrentasvir therapy: An integrated analysis of HCV genotype 1-6 patients without cirrhosis. J Hepatol. 2018;69(2):293–300. doi:.https://doi.org/10.1016/j.jhep.2018.03.007
  4. Aghemo A, Colombo M. Glecaprevir/Pibrentasvir: The Final Piece in the Hepatitis C Virus Treatment Puzzle? Gastroenterology. 2018;154(4):1195–6. doi:.https://doi.org/10.1053/j.gastro.2018.01.035
  5. Bregenzer A, Bruggmann P, Castro E, Moriggia A, Rothen M, Thurnheer MC, et al. Schweizer OAT-Programme auf ihrem Weg zur HCV-Elimination – die SAMMSU-Kohorte. Suchtmed. 2019;21(2):75–90.
  6. Bregenzer A, Conen A, Knuchel J, Friedl A, Eigenmann F, Näf M, et al. Management of hepatitis C in decentralised versus centralised drug substitution programmes and minimally invasive point-of-care tests to close gaps in the HCV cascade. Swiss Med Wkly. 2017;147:w14544.
  7. Janjua NZ, Kuo M, Yu A, Alvarez M, Wong S, Cook D, et al. The Population Level Cascade of Care for Hepatitis C in British Columbia, Canada: The BC Hepatitis Testers Cohort (BC-HTC). EBioMedicine. 2016;12:189–95. doi:.https://doi.org/10.1016/j.ebiom.2016.08.035
  8. Yehia BR, Schranz AJ, Umscheid CA, Lo Re V, 3rd. The treatment cascade for chronic hepatitis C virus infection in the United States: a systematic review and meta-analysis. PLoS One. 2014;9(7):e101554. doi:.https://doi.org/10.1371/journal.pone.0101554
  9. Hajarizadeh B, Grebely J, McManus H, Estes C, Razavi H, Gray RT, et al. Chronic hepatitis C burden and care cascade in Australia in the era of interferon-based treatment. J Gastroenterol Hepatol. 2017;32(1):229–36. doi:.https://doi.org/10.1111/jgh.13453
  10. Hayes B, Briceno A, Asher A, Yu M, Evans JL, Hahn JA, et al. Preference, acceptability and implications of the rapid hepatitis C screening test among high-risk young people who inject drugs. BMC Public Health. 2014;14(1):645. doi:.https://doi.org/10.1186/1471-2458-14-645
  11. Bottero J, Boyd A, Gozlan J, Carrat F, Nau J, Pauti MD, et al. Simultaneous Human Immunodeficiency Virus-Hepatitis B-Hepatitis C Point-of-Care Tests Improve Outcomes in Linkage-to-Care: Results of a Randomized Control Trial in Persons Without Healthcare Coverage. Open Forum Infect Dis. 2015;2(4):ofv162. doi:.https://doi.org/10.1093/ofid/ofv162
  12. Cooper C. Rapid HCV RNA testing: removing the final obstacle to elimination. Lancet Gastroenterol Hepatol. 2017;2(7):468–9. doi:.https://doi.org/10.1016/S2468-1253(17)30086-9
  13. Tang W, Chen W, Amini A, Boeras D, Falconer J, Kelly H, et al. Diagnostic accuracy of tests to detect Hepatitis C antibody: a meta-analysis and review of the literature. BMC Infect Dis. 2017;17(S1, Suppl 1):695. doi:.https://doi.org/10.1186/s12879-017-2773-2
  14. Ivanova Reipold E, Easterbrook P, Trianni A, Panneer N, Krakower D, Ongarello S, et al. Optimising diagnosis of viraemic hepatitis C infection: the development of a target product profile. BMC Infect Dis. 2017;17(S1, Suppl 1):707. doi:.https://doi.org/10.1186/s12879-017-2770-5
  15. Greenman J, Roberts T, Cohn J, Messac L. Dried blood spot in the genotyping, quantification and storage of HCV RNA: a systematic literature review. J Viral Hepat. 2015;22(4):353–61. doi:.https://doi.org/10.1111/jvh.12345
  16. Bennett S, Gunson RN, McAllister GE, Hutchinson SJ, Goldberg DJ, Cameron SO, et al. Detection of hepatitis C virus RNA in dried blood spots. J Clin Virol. 2012;54(2):106–9. doi:.https://doi.org/10.1016/j.jcv.2012.02.004
  17. McHugh MP, Wu AHB, Chevaliez S, Pawlotsky JM, Hallin M, Templeton KE. Multicenter Evaluation of the Cepheid Xpert Hepatitis C Virus Viral Load Assay. J Clin Microbiol. 2017;55(5):1550–6. doi:.https://doi.org/10.1128/JCM.02460-16
  18. Gupta E, Agarwala P, Kumar G, Maiwall R, Sarin SK. Point -of -care testing (POCT) in molecular diagnostics: Performance evaluation of GeneXpert HCV RNA test in diagnosing and monitoring of HCV infection. J Clin Virol. 2017;88:46–51. doi:.https://doi.org/10.1016/j.jcv.2017.01.006
  19. Cepheid®, Home → Cepheid Solutions → CE-IVD Tests → Virology → Xpert HCV Viral Load → Xpert HCV Viral Load Brochure - CE-IVD, http://www.cepheid.com/administrator/components/com_productcatalog/library-files/611eae732b51634d8a7e64878d8222a1-615085cee2245c0c5e547ebff5052119-Xpert-HCV-Viral-Load-Brochure-CEIVD-3043-02.pdf (Accessed: 16 Jan. 2018)
  20. Grebely J, Lamoury FMJ, Hajarizadeh B, Mowat Y, Marshall AD, Bajis S, et al.; LiveRLife Study Group. Evaluation of the Xpert HCV Viral Load point-of-care assay from venepuncture-collected and finger-stick capillary whole-blood samples: a cohort study. Lancet Gastroenterol Hepatol. 2017;2(7):514–20. doi:.https://doi.org/10.1016/S2468-1253(17)30075-4
  21. Lamoury FMJ, Bajis S, Hajarizadeh B, Marshall AD, Martinello M, Ivanova E, et al.; LiveRLife Study Group. Evaluation of the Xpert HCV Viral Load Finger-Stick Point-of-Care Assay. J Infect Dis. 2018;217(12):1889–96. doi:.https://doi.org/10.1093/infdis/jiy114
  22. Yan Y, Huang F, Yuan T, Sun B, Yang R. HIV-1 Vpr increases HCV replication through VprBP in cell culture. Virus Res. 2016;223:153–60. doi:.https://doi.org/10.1016/j.virusres.2016.07.007
  23. Lara J, Teka MA, Sims S, Xia GL, Ramachandran S, Khudyakov Y. HCV adaptation to HIV coinfection. Infect Genet Evol. 2018;65:216–25. doi:.https://doi.org/10.1016/j.meegid.2018.07.039
  24. Tuaillon E, Mondain AM, Meroueh F, Ottomani L, Picot MC, Nagot N, et al. Dried blood spot for hepatitis C virus serology and molecular testing. Hepatology. 2010;51(3):752–8.
  25. Santos C, Reis A, Dos Santos CV, Damas C, Silva MH, Viana MV, et al. The use of real-time PCR to detect hepatitis C virus RNA in dried blood spots from Brazilian patients infected chronically. J Virol Methods. 2012;179(1):17–20. doi:.https://doi.org/10.1016/j.jviromet.2011.06.012
  26. Stapleton JT, Klinzman D, Schmidt WN, Pfaller MA, Wu P, LaBrecque DR, et al. Prospective comparison of whole-blood- and plasma-based hepatitis C virus RNA detection systems: improved detection using whole blood as the source of viral RNA. J Clin Microbiol. 1999;37(3):484–9.
  27. Pham TN, MacParland SA, Mulrooney PM, Cooksley H, Naoumov NV, Michalak TI. Hepatitis C virus persistence after spontaneous or treatment-induced resolution of hepatitis C. J Virol. 2004;78(11):5867–74. doi:.https://doi.org/10.1128/JVI.78.11.5867-5874.2004
  28. Fourati S, Guedj J, Chevaliez S, Nguyen THT, Roudot-Thoraval F, Ruiz I, et al. Viral kinetics analysis and virological characterization of treatment failures in patients with chronic hepatitis C treated with sofosbuvir and an NS5A inhibitor. Aliment Pharmacol Ther. 2018;47(5):665–73. doi:.https://doi.org/10.1111/apt.14478
  29. Bajis S, Maher L, Treloar C, Hajarizadeh B, Lamoury FMJ, Mowat Y, et al.; LiveRLife Study Group. Acceptability and preferences of point-of-care finger-stick whole-blood and venepuncture hepatitis C virus testing among people who inject drugs in Australia. Int J Drug Policy. 2018;61:23–30. doi:.https://doi.org/10.1016/j.drugpo.2018.08.011
  30. Bajis S, Grebely J, Cooper L, Smith J, Owen G, Chudleigh A, et al. Hepatitis C virus testing, liver disease assessment and direct-acting antiviral treatment uptake and outcomes in a service for people who are homeless in Sydney, Australia: The LiveRLife homelessness study. J Viral Hepat. 2019;26(8):969–79. doi:.https://doi.org/10.1111/jvh.13112
  31. Wlassow M, Poiteau L, Roudot-Thoraval F, Rosa I, Soulier A, Hézode C, et al. The new Xpert HCV viral load real-time PCR assay accurately quantifies hepatitis C virus RNA in serum and whole-blood specimens. J Clin Virol. 2019;117:80–4. doi:.https://doi.org/10.1016/j.jcv.2019.06.007
  32. Ahmed M, Munshi SU, Andalib S, Tabassum S, Islam MN. Testing Hepatitis A virus antibody in oral fluid among the prospective vaccinees foster the need of new oral HAV rapid test. Indian J Med Microbiol. 2011;29(1):72–3. doi:.https://doi.org/10.4103/0255-0857.76532
  33. de Almeida LM, Azevedo RS, Marandino Guimarães AA, da Silva Freire Coutinho E, Struchiner CJ, Massad E. Detection of antibodies against hepatitis A virus in eluates of blood spotted on filter-paper: a pilot study in Rio de Janeiro, Brazil. Trans R Soc Trop Med Hyg. 1999;93(4):401–4. doi:.https://doi.org/10.1016/S0035-9203(99)90133-5
  34. Shivkumar S, Peeling R, Jafari Y, Joseph L, Pai NP. Rapid point-of-care first-line screening tests for hepatitis B infection: a meta-analysis of diagnostic accuracy (1980-2010). Am J Gastroenterol. 2012;107(9):1306–13. doi:.https://doi.org/10.1038/ajg.2012.141
  35. El-Ghitany EM, Farghaly AG. Evaluation of commercialized rapid diagnostic testing for some Hepatitis B biomarkers in an area of intermediate endemicity. J Virol Methods. 2013;194(1-2):190–3. doi:.https://doi.org/10.1016/j.jviromet.2013.08.026
  36. Wu FY, Liao YW, Wu JF, Chen HL, Hsu HY, Chang MH, et al. A simple and rapid test-card method to detect hepatitis B surface antigen and antibody: potential application in young children and infants. Pediatr Neonatol. 2016;57(3):219–24. doi:.https://doi.org/10.1016/j.pedneo.2015.07.003
  37. Poiteau L, Soulier A, Roudot-Thoraval F, Hézode C, Challine D, Pawlotsky JM, et al. Performance of rapid diagnostic tests for the detection of anti-HBs in various patient populations. J Clin Virol. 2017;96:64–6. doi:.https://doi.org/10.1016/j.jcv.2017.09.012

Most read articles by the same author(s)