HIV – Diagnostic Serology

Conformément au Règlement de l’Ontario 671/92 de la Loi sur les services en français, les renseignements d’analyses de laboratoire liés à la présente page ne sont offerts qu’en anglais parce qu’ils sont de nature scientifique ou technique et destinés uniquement à l’usage des fournisseurs de soins de santé qualifiés et non aux membres du public.

This page provides diagnostic serology testing information for HIV at Public Health Ontario (PHO). For information regarding other testing options, refer to the following PHO webpages:

* Copie d'impression non contrôlée. Valide uniquement le jour de l'impression : 19 avr. 2024.

Testing Indications

HIV testing should be considered in individuals:

with symptoms of acute HIV infection
with symptoms associated with chronic HIV infection
who have had a high-risk exposure
who belong to a population with higher rates of HIV
who are initiating immune suppressing therapy
who are pregnant; if ordering as part of a prenatal screening, see Prenatal – Serology

For more information on HIV testing recommendations, refer to the Ontario Guidelines for Providers Offering HIV Testing.

Acceptance/Rejection Criteria

Donor testing is not available through PHO’s laboratory. Specimens from patients being screened as potential donors (e.g. organ, tissue, cells, fertility, etc.) should be referred to a laboratory that performs donor screening assays. Specimens received for donor screening at PHO’s laboratory will be rejected.
Specimens received more than 7 days post collection will not be tested.

Specimen Collection and Handling

Specimen Requirements

Test Requested	Required Requisition(s)	Specimen Type	Minimum Volume	Collection Kit
HIV	HIV Serology and HIV PCR Test Requisition	Serum	1.5 ml	Serum Separator Tube (SST)

Submission and Collection Notes

Complete all fields of the requisition form, including:

Test(s) requests and reason for testing
Patient setting, specimen type and site
Relevant clinical information

Label the specimen container(s) with the patient’s first and last name, date of collection, and one other unique identifier such as the patient’s date of birth or health card number. For additional information see: Criteria for Acceptance of Patient Specimens. Failure to provide this information may result in rejection or testing delay.

Submit centrifuged serum separator tubes (SST) for serological assays.

One FULL 5 mL SST is required for testing a combination of HIV, hepatitis virus, HTLV, syphilis and rubella.

Do not submit glass tubes.

Limitations

Haemolysed, icteric, lipemic or microbially contaminated sera or plasma are not recommended for testing.

Storage and Transport

Place specimen tube in biohazard bag and seal. Place completed General Test Requisition in the pouch at the front of the biohazard bag.

Specimen tubes should be stored at 2-8°C following collection and centrifugation, and should be shipped to PHO’s laboratory on ice packs within 3 days of collection. If delayed shipping is anticipated, remove serum from clot and store frozen at -20°C or colder, and ship on dry ice.

All clinical specimens must be shipped in accordance to the Transportation of Dangerous Good Act.

Requisitions and Kit Ordering

Test Frequency and Turnaround Time (TAT)

HIV diagnostic serology testing is performed daily Monday to Friday.

Turnaround time is up to 3 business days for non-reactive specimens and up to 6 business days for reactive specimens from receipt by PHO’s laboratory.

Stat and Critical Samples Testing

STAT testing should only be requested when it directly affects patient care in an emergency medical circumstance. If warranted, the following HIV testing is expedited:

Testing of the ‘source patient’ for needlestick/occupational exposure
Pregnant women presenting in labour who have not received prenatal care or for whom results of previous testing are not readily available and are a high risk for HIV

STAT samples must be shipped separately from routine specimens and delivered to PHO’s laboratory in a timely manner. The outer package must be clearly marked indicating ‘STAT’. It must be handled in accordance with the Canadian Biosafety Standards and shipped in accordance with the Transportation of Dangerous Goods Regulations. Failure to ship separately will delay testing.

Prior to sending, notify PHO’s laboratory Customer Service Centre at 416-235-6556 / 1-877-604-4567. After-hours STAT testing requires approval by a PHO Microbiologist. Contact the After-Hours Emergency Duty Officer at 416-605-3113 if outside of PHO’s laboratory operating hours.

Test Methods

Specimens are screened for human immunodeficiency virus (HIV) using a chemiluminescent microparticle immunoassay (CMIA) for the simultaneous qualitative detection of HIV-1 p24 antigen (Ag) and antibodies to HIV-1 and HIV-2, but does not differentiate between the two.

If the CMIA test is reactive or indeterminate, the specimen is tested using the Geenius™ HIV-1/-2 Differential assay, which is an immunochromatographic test for the confirmation and differentiation of antibodies to HIV-1 (Group M and O) and HIV-2.

CMIA-positive samples that are negative or indeterminate by the Geenius™ HIV-1/-2 Differential assay are screened for p24 Ag using an Enzyme Linked Fluorescent Assay (ELFA) for the detection of HIV-1 p24 Ag. Samples that are positive for p24 Ag are confirmed using a p24 Ag neutralization assay.

Interpretation

Results should be interpreted in the context of the patient’s clinical presentation, risk factors, and exposure history. The table below provides a general overview of the algorithm and most common interpretations for HIV screening and confirmatory testing. It does not include all possible result scenarios. A PHO Microbiologist is available for questions about HIV serology results and interpretations.

HIV Ab/Ag CMIA Screen	Geenius^TM Confirmatory Assay	p24 Ag ELFA	Interpretation
Non-Reactive	Not Tested	Not Tested	No HIV p24 antigen and no HIV-1/HIV-2 antibodies detected.
Reactive/ Indeterminate	Positive	Not Tested	HIV-1 antibodies detected OR HIV-2 antibodies detected
Reactive/ Indeterminate	Negative/ Indeterminate	Reactive	Evidence of HIV-1 infection prior to seroconversion.
Reactive/ Indeterminate	Negative/ Indeterminate	Non-Reactive/ Indeterminate	Inconclusive HIV p24 antigen and inconclusive HIV-1/HIV-2 antibodies. Submit a follow-up specimen for repeat testing at least 4 weeks after collection of the first specimen.

Reporting

Results are reported to the physician, authorized health care provider (General O. Reg 45/22, s.18) or submitter as indicated on the requisition.

Specimens that are positive for HIV diagnostic testing are reported to the Medical Officer of Health as per the Ontario Health Protection and Promotion Act.