What hepatitis B (hep B) serology tests should I request?

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Last updated: November 20, 2025View editorial policy

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Hepatitis B Serology Testing

Order a three-test panel consisting of hepatitis B surface antigen (HBsAg), total or IgG antibody to hepatitis B core antigen (anti-HBc), and antibody to hepatitis B surface antigen (anti-HBs) for comprehensive hepatitis B screening. 1

Core Testing Panel

The standard hepatitis B serology panel includes three essential markers that together distinguish between active infection, resolved infection, immunity from vaccination, and susceptibility 2:

  • HBsAg (Hepatitis B Surface Antigen): Detects active infection (both acute and chronic). This is the hallmark marker of HBV infection and the first to appear, typically within 30 days of exposure (range 6-60 days) 2

  • Anti-HBc total or IgG (Antibody to Hepatitis B Core Antigen): Indicates previous or ongoing infection. This marker persists for life in the majority of persons after HBV exposure and distinguishes natural infection from vaccine-induced immunity 2, 1

  • Anti-HBs (Antibody to Hepatitis B Surface Antigen): Indicates immunity from either vaccination or resolved infection 2, 1

Interpretation Algorithm

The combination of these three tests provides definitive diagnostic information 1, 3:

HBsAg Anti-HBc Anti-HBs Interpretation
Positive Positive Negative Active chronic HBV infection
Negative Positive Positive Resolved past infection
Negative Negative Positive Immunity from vaccination
Negative Positive Negative Isolated core antibody (possible occult HBV)
Negative Negative Negative Susceptible (needs vaccination)

Additional Testing When HBsAg is Positive

If HBsAg is positive, immediately order HBeAg, anti-HBe, and quantitative HBV DNA to assess disease activity and guide treatment decisions. 2, 1

  • HBeAg positivity typically indicates high viral replication and increased infectivity 2
  • Quantitative HBV DNA is essential for determining treatment eligibility and monitoring response 2
  • ALT levels should be assessed to evaluate liver inflammation 2

Additional Testing for Isolated Anti-HBc

If the pattern shows isolated anti-HBc positivity (HBsAg negative, anti-HBc positive, anti-HBs negative), order quantitative HBV DNA testing to rule out occult HBV infection, particularly in immunocompromised patients or those with unexplained elevated aminotransferases 2, 1, 3

Critical Pitfalls to Avoid

  • Do NOT order only HBsAg: This single test will miss resolved infections, vaccine immunity status, and isolated core antibody patterns 1

  • Do NOT order IgM anti-HBc for routine screening: Reserve this test exclusively for suspected acute hepatitis B (clinical symptoms or epidemiologic link to infection). The positive predictive value is low in asymptomatic persons 2

  • Do NOT assume vaccination history eliminates testing need: Breakthrough infections can occur, and documented vaccination history is often unreliable 1

  • Do NOT interpret isolated anti-HBc definitively without HBV DNA testing: This pattern requires further evaluation as it may represent occult infection, particularly in immunocompromised patients 2, 1, 3

  • Do NOT delay testing in patients starting immunosuppressive therapy or chemotherapy: Complete serologic assessment is essential before initiating these treatments to assess reactivation risk, but do not delay cancer therapy while awaiting results 1

Special Clinical Contexts

For patients with chronic liver disease from other causes (such as hepatitis C), the same three-test panel is recommended to determine if hepatitis B vaccination is needed, as coinfection significantly increases morbidity and mortality 4

For healthcare workers or those with occupational exposure, testing both anti-HBs and anti-HBc distinguishes vaccine-induced immunity from infection-induced immunity 3

References

Guideline

Hepatitis B Testing and Interpretation

Praxis Medical Insights: Practical Summaries of Clinical Guidelines, 2025

Guideline

Guideline Directed Topic Overview

Dr.Oracle Medical Advisory Board & Editors, 2025

Guideline

Hepatitis A and B Titer Testing Guidelines

Praxis Medical Insights: Practical Summaries of Clinical Guidelines, 2025

Professional Medical Disclaimer

This information is intended for healthcare professionals. Any medical decision-making should rely on clinical judgment and independently verified information. The content provided herein does not replace professional discretion and should be considered supplementary to established clinical guidelines. Healthcare providers should verify all information against primary literature and current practice standards before application in patient care. Dr.Oracle assumes no liability for clinical decisions based on this content.

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