Hepatitis C Virus - HCV

Hepatitis C virus (HCV) is a virally mediated disease of the liver with a propensity to cause chronic infection, leading to cirrhosis and an increased risk of hepatocellular carcinoma. Adults born between 1945 and 1965 should be screened for HCV infection; individuals with certain risk behaviors (eg, injection drug use) or risk exposures (eg, healthcare workers) should also be screened. Laboratory testing involves screening for HCV antibodies followed by confirmatory RNA testing for positive results.

  • Diagnosis
  • Algorithms
  • Screening
  • Monitoring
  • Background
  • Lab Tests
  • References
  • Related Topics
  • Videos

Indications for Testing

  • New onset of jaundice, anorexia, or dark urine
  • Elevated liver enzymes
  • Known or suspected exposure to hepatitis C virus (HCV)
  • Liver fibrosis or cirrhosis without identified etiology

Laboratory Testing

  • See Screening section for information concerning at-risk individuals/those who should be screened
  • Testing recommendations for HCV (CDC)
    • Initial testing
      • HCV serology screen by chemiluminescence immunoassay (CIA), enzyme immunoassay (EIA)
        • Use for initial testing in patient with suspected HCV
        • High rate of false positives and positive serology due to past (inactive) infection – confirmatory testing for active infection by HCV RNA is required
    • Follow-up testing
      • HCV RNA
        • Indications
          • Positive HCV serology test – confirm active infection
          • Immunocompromised individuals – serology is dependent on ability to mount immune response; HCV RNA should be first test in these patients
          • HCV exposure in previous 6 months – test for RNA, not serology
          • Concern for reinfection following successful HCV treatment
          • Prior to start of treatment – document baseline RNA elevation
        • Tests
          • Quantitative
            • Provides information about RNA viral load
          • Qualitative
            • Less expensive than quantitative; however, in most cases quantification is needed for purposes other than confirmation, so quantitative testing is preferred over qualitative
      • HCV genotyping
        • Indications
          • Guide selection of direct acting antiviral (DAA) therapy
        • Tests
          • Polymerase chain reaction (PCR) by Sanger – may be sufficient to determine genotype and direct therapy
          • Next generation sequencing, other deep-sequencing tests – sometimes indicated if further genotyping is required
      • Resistance-associated variant (RAV) testing – NS5A
        • Not appropriate to perform prior to genotype testing
        • Recommended at baseline for patients with HCV genotype 1a prior to initiation of treatment with NS5A inhibitors (eg, elbasvir plus grazoprevir)
        • Consider for patients with genotype 1a and cirrhosis prior to treatment with sofosbuvir plus daclatasvir


  • Genotype influences response to therapy
  • Viral load (as measured by quantitative PCR) predicts likelihood of treatment response
  • Coinfection with hepatitis B (HBV) predicts poorer prognosis
  • Presence or absence of cirrhosis (stable or decompensated) will influence response
  • With advent of new, highly effective therapies, compliance may be most influential determinant

Differential Diagnosis

  • Screening indications
    • Date of birth between 1945-1965 – test at least once (American Association for the Study of Liver Diseases [AASLD]/Infectious Diseases Society of America [IDSA], 2017; Smith, CDC, 2012; Moyer, U.S. Preventive Services Task Force [USPSTF], 2013)
    • Injection drug use or HIV infection in men who have unprotected sex with men – test annually
    • Risk behaviors or exposures for hepatitis C (HCV) infection (see AASLD/IDSA recommendations, 2017)  – test once
      • Risk behaviors
        • Injection drug use (current or ever, including 1-time use)
        • Intranasal illicit drug use
      • Risk exposures
        • Patients with long-term hemodialysis (ever)
        • Healthcare, emergency medical, and public safety workers after needlestick, sharps, or mucosal exposure to HCV-infected blood
        • Babies born to HCV-infected mothers
        • Prior recipients of transfusions, including individuals who
          • Were notified that they received blood from a donor who later tested positive for HCV infection
          • Received a transfusion of blood or blood components, or underwent an organ transplant before July 1992
          • Received clotting factor concentrates produced before 1987
          • Have been incarcerated
          • Are HIV positive
      • Other medical conditions
        • Unexplained chronic liver disease and chronic hepatitis, including elevated alanine aminotransferase levels
  • Laboratory testing (refer to Diagnosis section)
    • Initial screening for HCV antibodies by chemiluminescence immunoassay (CIA), enzyme immunoassay (EIA), enzyme-linked immunosorbent assay (ELISA)
    • Confirmatory RNA testing required for positive result
  • Hepatitis C virus (HCV) RNA quantitative polymerase chain reaction (PCR) test
    • Establish viral load at baseline
    • Evaluate response to therapy at 4 weeks
    • Reevaluate at end of prescribed duration of therapy


  • Prevalence – estimated 2.7-3.9 million infected in the U.S. (CDC)
    • >50% of new cases are caused by intravenous drug use; majority of previous U.S. cases were attributed to transfusion of infected blood products
  • Sex – M:F, equal


  • Single-stranded RNA virus; member of Flaviviridae family (genus Hepacivirus)
  • Multiple genotypes with multiple subtypes (1a, 1b, 1c, etc.)
    • Genotype is an important predictor of virological response to hepatitis C virus (HCV) treatment
      • Type 1 is predominant genotype in U.S. and more difficult to treat
      • Clinically, type 1 subtypes (1a and 1b) are most relevant

Risk Factors

Refer to Screening section

Clinical Presentation

  • HCV is typically asymptomatic as an acute infection
    • Infection may be initially identified when patient has positive anti-HCV in a blood donor screen or has high alanine aminotransferase level (10-20 times the upper limit of normal) in blood chemistry testing for flu-like symptoms
  • Chronic disease occurs in ~10-20% of patients
    • Cirrhosis (20%) and hepatocellular carcinoma (1-5%)
  • Chronic asymptomatic hepatitis may manifest with other systemic symptoms
    • Mixed cryoglobulinemia – systemic vasculitis involving skin, kidneys, nervous system
    • Sjögren syndrome – anti-Sjögren syndrome A (anti-SSA) and anti-Sjögren syndrome B (anti-SSB) antibodies are usually absent or are present at low levels
    • Lichen planus – violaceous papules on any skin site; oral most common
    • Porphyria cutanea tarda
    • Non-Hodgkin lymphoma – B-cell type most common
  • Pregnant women
    • Not transmitted to infant via breastfeeding 
    • Pregnancy not contraindicated
Tests generally appear in the order most useful for common clinical situations. Click on number for test-specific information in the ARUP Laboratory Test Directory.

Hepatitis C Virus Antibody by CIA with Reflex to HCV by Quantitative PCR 2010784
Method: Qualitative Chemiluminescent Immunoassay/Quantitative Polymerase Chain Reaction

Hepatitis C Virus Antibody by CIA 2002483
Method: Qualitative Chemiluminescent Immunoassay

Hepatitis C Virus (HCV) by Quantitative PCR with Reflex to HCV Genotype by Sequencing 2002685
Method: Quantitative Polymerase Chain Reaction/Sequencing


If virus is not detected, result will be reported as <1.2 log IU/mL; if virus is detected but number of copies is not accurately quantified, result will be reported as not quantified

Hepatitis C Virus by Quantitative PCR 0098268
Method: Quantitative Polymerase Chain Reaction


If virus is not detected, result will be reported as <1.2 log IU/mL; if virus is detected but number of copies is not accurately quantified, result will be reported as not quantified

Hepatitis C Virus Genotype by Sequencing 0055593
Method: Polymerase Chain Reaction/Sequencing


Due to high conservation of the 5' untranslated region of the HCV genome, this test has limitations in differentiating subtype 1a from 1b; therefore, these subtypes will be reported as "1a or 1b"

In rare instances, type 6 virus may be misclassified as type 1

Test may be unsuccessful if HCV RNA viral load is <log 3.6 or 4,000 IU/mL

Hepatitis C Virus (HCV) by Quantitative PCR with Reflex to HCV High-Resolution Genotype by Sequencing 2010793
Method: Quantitative Polymerase Chain Reaction/Sequencing


If virus is not detected, result will be reported as <1.2 log IU/mL; if virus is detected but number of copies is not accurately quantified, result will be reported as not quantified

Hepatitis C Virus (HCV) Genotype with Reflex to HCV High-Resolution Genotype by Sequencing 2009255
Method: Polymerase Chain Reaction/Sequencing


Test may be unsuccessful if HCV RNA viral load is <log 5.0 or 100,000 IU/mL

Interleukin 28 B (IL28B)-Associated Variants, 2 SNPs 2004680
Method: Polymerase Chain Reaction/Single Nucleotide Extension


Single nucleotide polymorphisms (SNPs) other than those targeted will not be detected

Usefulness of IL28B-associated SNPs for predicting therapy response for HCV genotypes other than HCV-1 is unknown

Other gene variants and nongenetic factors that may affect response to HCV therapy are not detected

Diagnostic errors can occur due to rare sequence variations


Easterbrook PJ, WHO Guidelines Development Group. Who to test and how to test for chronic hepatitis C infection - 2016 WHO testing guidance for low- and middle-income countries. J Hepatol. 2016; 65(1 Suppl): S46-S66. PubMed

European Association for Study of Liver. EASL Clinical Practice Guidelines: management of hepatitis C virus infection. J Hepatol. 2014; 60(2): 392-420. PubMed

Full Report: HCV Guidance: Recommendations for Testing, Managing, and Treating Hepatitis C. American Association for the Study of Liver Diseases; Infectious Diseases Society of America. [Revised: Apr 2017; Accessed: Jun 2017]

Ghany MG, Nelson DR, Strader DB, Thomas DL, Seeff LB, American Association for Study of Liver Diseases. An update on treatment of genotype 1 chronic hepatitis C virus infection: 2011 practice guideline by the American Association for the Study of Liver Diseases. Hepatology. 2011; 54(4): 1433-44. PubMed

Guidelines for the Screening, Care and Treatment of Persons with Hepatitis C Infection. World Health Organization. Geneva, Switzerland [Accessed: Jun 2017]

HCV Guidance: Recommendations for Testing, Managing, and Treating Hepatitis C. AASLD/IDSA HCV Guidance Panel. [Updated/Changes: Apr 2017; Accessed: Jun 2017]

Moyer VA, U.S. Preventive Services Task Force. Screening for hepatitis C virus infection in adults: U.S. Preventive Services Task Force recommendation statement. Ann Intern Med. 2013; 159(5): 349-57. PubMed

Ramos-Casals M, Zignego AL, Ferri C, Brito-Zerón P, Retamozo S, Casato M, Lamprecht P, Mangia A, Saadoun D, Tzioufas AG, Younossi ZM, Cacoub P, International Study Group of Extrahepatic Manifestations related to HCV (ISG-EHCV). Evidence-based recommendations on the management of extrahepatic manifestations of chronic hepatitis C virus infection. J Hepatol. 2017; 66(6): 1282-1299. PubMed

Smith B, Morgan R, Beckett G, et al. Recommendations for the Identification of Chronic Hepatitis C Virus Infection Among Persons Born During 1945–1965. Centers for Disease Control and Prevention. Atlanta, GA [Updated: Aug 2012; Accessed: Jun 2017]

General References

Ahlenstiel G, Booth DR, George J. IL28B in hepatitis C virus infection: translating pharmacogenomics into clinical practice. J Gastroenterol. 2010; 45(9): 903-10. PubMed

Albeldawi M, Ruiz-Rodriguez E, Carey WD. Hepatitis C virus: Prevention, screening, and interpretation of assays. Cleve Clin J Med. 2010; 77(9): 616-26. PubMed

Balagopal A, Thomas DL, Thio CL. IL28B and the control of hepatitis C virus infection. Gastroenterology. 2010; 139(6): 1865-76. PubMed

Chakravarty R. Diagnosis and monitoring of chronic viral hepatitis: serologic and molecular markers. Front Biosci (Schol Ed). 2011; 3: 156-67. PubMed

Chan J. Hepatitis C. Dis Mon. 2014; 60(5): 201-12. PubMed

Chevaliez S, Hézode C. IL28B polymorphisms and chronic hepatitis C. Gastroenterol Clin Biol. 2010; 34(11): 587-9. PubMed

Cobb B, Heilek G, Vilchez RA. Molecular diagnostics in the management of chronic hepatitis C: key considerations in the era of new antiviral therapies. BMC Infect Dis. 2014; 14 Suppl 5: S8. PubMed

Gullett JC, Nolte FS. Quantitative nucleic acid amplification methods for viral infections. Clin Chem. 2015; 61(1): 72-8. PubMed

Maheshwari A, Thuluvath PJ. Management of acute hepatitis C. Clin Liver Dis. 2010; 14(1): 169-76; x. PubMed

Morrison BJ, Labo N, Miley WJ, Whitby D. Serodiagnosis for tumor viruses. Semin Oncol. 2015; 42(2): 191-206. PubMed

Pearlman BL. The IL-28 genotype: how it will affect the care of patients with hepatitis C virus infection. Curr Gastroenterol Rep. 2011; 13(1): 78-86. PubMed

Sagnelli E, Santantonio T, Coppola N, Fasano M, Pisaturo M, Sagnelli C. Acute hepatitis C: clinical and laboratory diagnosis, course of the disease, treatment. Infection. 2014; 42(4): 601-10. PubMed

Viral Hepatitis - Hepatitis C Information - Testing Recommendations. Centers for Disease Control and Prevention. Atlanta, GA [Accessed: Dec 2016]

Webster DP, Klenerman P, Dusheiko GM. Hepatitis C. Lancet. 2015; 385(9973): 1124-35. PubMed

Wilkins T, Malcolm JK, Raina D, Schade RR. Hepatitis C: diagnosis and treatment. Am Fam Physician. 2010; 81(11): 1351-7. PubMed

References from the ARUP Institute for Clinical and Experimental Pathology®

Bossler A, Gunsolly C, Pyne MT, Rendo A, Rachel J, Mills R, Miller M, Sipley J, Hillyard D, Jenkins S, Essmyer C, Young S, Lewinski M, Rennert H. Performance of the COBAS® AmpliPrep/COBAS TaqMan® automated system for hepatitis C virus (HCV) quantification in a multi-center comparison. J Clin Virol. 2011; 50(2): 100-3. PubMed

Konnick EQ, Erali M, Ashwood ER, Hillyard DR. Performance characteristics of the COBAS Amplicor Hepatitis C Virus (HCV) Monitor, Version 2.0, International Unit assay and the National Genetics Institute HCV Superquant assay. J Clin Microbiol. 2002; 40(3): 768-73. PubMed

Konnick EQ, Williams SM, Ashwood ER, Hillyard DR. Evaluation of the COBAS Hepatitis C Virus (HCV) TaqMan analyte-specific reagent assay and comparison to the COBAS Amplicor HCV Monitor V2.0 and Versant HCV bDNA 3.0 assays. J Clin Microbiol. 2005; 43(5): 2133-40. PubMed

Liew M, Erali M, Page S, Hillyard D, Wittwer C. Hepatitis C genotyping by denaturing high-performance liquid chromatography. J Clin Microbiol. 2004; 42(1): 158-63. PubMed

Mallory MA, Lucic DX, Sears MT, Cloherty GA, Hillyard DR. Evaluation of the Abbott realtime HCV genotype II RUO (GT II) assay with reference to 5'UTR, core and NS5B sequencing. J Clin Virol. 2014; 60(1): 22-6. PubMed

Margraf RL, Erali M, Liew M, Wittwer CT. Genotyping hepatitis C virus by heteroduplex mobility analysis using temperature gradient capillary electrophoresis. J Clin Microbiol. 2004; 42(10): 4545-51. PubMed

Margraf RL, Page S, Erali M, Wittwer CT. Single-tube method for nucleic acid extraction, amplification, purification, and sequencing. Clin Chem. 2004; 50(10): 1755-61. PubMed

Melis R, Fauron C, McMillin G, Lyon E, Shirts B, Hubley LM, Slev PR. Simultaneous genotyping of rs12979860 and rs8099917 variants near the IL28B locus associated with HCV clearance and treatment response. J Mol Diagn. 2011; 13(4): 446-51. PubMed

Pyne MT, Hillyard DR. Evaluation of the Roche COBAS AmpliPrep/COBAS TaqMan HCV Test. Diagn Microbiol Infect Dis. 2013; 77(1): 25-30. PubMed

Pyne MT, Konnick EQ, Phansalkar A, Hillyard DR. Evaluation of the Abbott investigational use only RealTime hepatitis C virus (HCV) assay and comparison to the Roche TaqMan HCV analyte-specific reagent assay. J Clin Microbiol. 2009; 47(9): 2872-8. PubMed

Pyne MT, Mallory M, Hillyard DR. HCV RNA measurement in samples with diverse genotypes using versions 1 and 2 of the Roche COBAS® AmpliPrep/COBAS® TaqMan® HCV test. J Clin Virol. 2015; 65: 54-7. PubMed

Slev P. Host genomics and HCV personalized medicine. Ann Clin Lab Sci. 2012; 42(4): 363-9. PubMed

Medical Reviewers

Content Reviewed: 
May 2017

Last Update: February 2018