Hepatitis B – Serology

Testing Indications

Testing for hepatitis serology may be indicated during the work-up patients with suspected acute or chronic hepatitis, as part of pre-natal screening, to determine immunity (either following recovery from natural infection or as a result of immunization), and for epidemiologic purposes (e.g. investigation of infection control breach).

If ordering as part of a Prenatal Serology work-up, see Prenatal – Serology.

Specimen Collection and Handling

Specimen Requirements

Test Requested Required Requisition(s) Specimen Type Minimum Volume Collection Kit

Hepatitis B1


Immunity2: 1.0 mL  

Diagnostic2: 3.5 mL

Blood, clotted - vacutainer tubes (SST)

Submission and Collection Notes


Detailed instructions for ordering Hepatitis B testing are available to assist with completing the “Hepatitis Serology boxes” in section 3 of the General Test Requisition. 


For testing all Hepatitis markers, HIV, HTLV, Syphilis and Rubella, you only are required to submit two FULL red top or serum separator tubes (SST).


Do NOT submit glass tubes.


Haemolysed, icteric, lipemic or microbially contaminated serum is not recommended for testing.

Storage and Transport

Label the specimen container with the patient’s full name, date of collection and one other unique identifier such as the patient’s date of birth or Health Card Number. Failure to provide this information may result in rejection or testing delay.

Centrifuge if using SST. Place specimen in biohazard bag and seal. Specimens should be stored at 2-8°C following collection.

Specimens more than the following number of days post collection will not be tested:

  • >6 days for Hepatitis B surface antigen [HBsAg]
  • >7 days for Hepatitis B e Antigen and Hepatitis B e Antibody
  • > 10 days for Hepatitis B core Antigen and Hepatitis B surface Antibody

Special Instructions

Instructions for using SST tubes are found in the document titled: LAB-SD-008, Blood Collection using Serum Separator Tubes

Requisitions and Kit Ordering

Test Frequency and Turnaround Time (TAT)

Hepatitis B - Serology testing is performed daily Monday to Friday.

Turnaround time is up to 3 days from receipt by PHO laboratory for non-reactive samples and up to 6 days for Reactive samples.

STAT and Critical Samples Testing

HBsAg - Testing on ‘Source Patient’ for Needlestick/Occupational Exposure if warranted

Test Methods

Specimens requesting Hepatitis B serology  are tested  using a chemiluminescent microparticle immunoassay (CMIA) for the qualitative detection of Hepatitis B markers (either antibodies or antigens).

If HBsAg is reactive, further testing for Anti-HBc (Total) is performed automatically.

If HBsAg is reactive, but the anti-HBc (total) is non-reactive then the specimen is tested using a HBsAg confirmatory test (‘blocking assay’).

If the HBsAg and anti-HBc (total) are both reactive and the patient has acute symptoms indicated on the requisition, then the specimen will have the following tests performed: HBeAg, anti-HBe and anti-HBc IgM.


Please refer to: https://www.cdc.gov/hepatitis/hbv/profresourcesb.htm for interpretation of hepatitis B serologic test results.


Results are reported to the ordering physician or health care provider as indicated on the requisition.

Specimens that are positive for Hepatitis B – HbsAg are to be reported to the Medical Officer of Health as per Health Protection and Promotion Act.

Mis à jour le 2 oct. 2020