Mycobacterium – CSF, Fluids, Bone or Tissue

Consistent with O. Reg. 671/92 of the French Language Services Act, laboratory testing information on this page is only available in English because it is scientific or technical in nature and is for use only by qualified health care providers and not by members of the public.

Specimen Collection and Handling

Specimen Requirements

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

Mycobacterium culture

CSF

0.5 ml

Tuberculosis Kit order#: 390042

Mycobacterium culture

Fluids

5.0 ml

Tuberculosis Kit order#: 390042

Mycobacterium culture

Tissue or bone

1-2 grams

Tuberculosis Kit order#: 390042

Submission and Collection Notes

1

Use the Tuberculosis kit to submit specimens collected aseptically. Specimens in fixative will not be tested.

2

Small volumes of fluid may be submitted in a syringe with needle removed and Luer Lock cap in place.

3

Collect tissue using aseptic technique, do not wrap in gauze, select caseous portion if available, and do not freeze.

4

If submitting tissue from lymph nodes, place a small amount of sterile saline in the kit and do not freeze.

5

Multiple tissues submitted from the same surgical site should be limited to 3 specimens. If more than 3 specimens are required for testing, please contact the laboratory.

6

The following samples will not be tested: Specimens submitted in ANAEROBIC TRANSPORT MEDIUM, specimens received in formalin or as dry swabs, frozen specimens and CSF specimens <0.5 ml.

7

CSF volume equal to 0.5mL will have culture performed only (no smear). Please submit maximum volume attainable for culture and smear.

8

Refer to Additional Mycobacterium Collection Information for detailed collection instructions.

Limitations

Swabs are not recommended for the isolation of mycobacteria since they provide limited material and the bacteria adsorb onto the swab material which may contain toxic substances. Negative results obtained from specimens submitted on swabs are not reliable.

Frozen specimens may decrease the yield of mycobacteria.

Specimens obtained for initial diagnosis after the initiation of antimicrobial therapy may produce false negative results.

Saprophytic mycobacteria in tap water may produce false-positive culture or slide results.

Delayed transportation of specimens may promote overgrowth by contaminating indigenous microbiota.

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.

Special Instructions

Store at 2-8 °C (CSF at room temperature) after collection. Specimens should be transferred to the lab ASAP.

Requisitions and Kit Ordering

Test Frequency and Turnaround Time (TAT)

Mycobacterium cultures are performed daily Monday to Saturday. 

Turnaround time is up to 1 day from receipt by PHO laboratory for smear results from the concentrated specimen.

Turnaround time for negative cultures is 49 days from receipt by PHO laboratory.

A positive culture is reported within 24 hours of growth. Depending on the species and treatment of the patient, a culture may grow within 1 to 2 weeks or take as long as 6-7 weeks. All new isolates of M. tuberculosis complex are phoned to the submitter within 24 hours of identification.

STAT and Critical Samples Testing

STAT testing is not available.

Contact Public Health Ontario laboratory to request approval for URGENT testing.

If URGENT testing is approved, mark URGENT on the outside of the package as well as on the requisition. A phone number and contact person must be provided for phoning the result.

URGENT Smears Specimens that are received in the lab by 2:00 p.m. will have a smear done and phoned the same day. Specimens that arrive after 2 p.m. will have the smear result phoned first thing the following morning.

Test Methods

A smear, prepared from the concentrate, is stained using the Auramine-Rhodamine (AR) fluorochrome stain and fluorescence microscopy.

A negative AFB smear does not rule out TB, the culture may still be positive.

Specimens are set up for culture on solid and in liquid media and incubated for up to 7 weeks.

Reporting

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

Specimens that are positive for Mycobacterium tuberculosis are to be reported to the Medical Officer of Health as per Health Protection and Promotion Act.

Additional Collection Information

Additional Mycobacterium Collection Information

Updated 26 July 2022