Identification of Non-tuberculous Mycobacterium species (NTM)

Identification of Non-tuberculous Mycobacterium spp to the species level as per ‘LPSN (List of Prokaryotic names with Standing in Nomenclature, author:  J.P. Euzeby).’  The testing is conducted using a combination of MALDI-TOF (matrix-assisted laser desorption/ionization – Time Of Flight) Mass Spectrometry* and/or DNA sequence-based methodologies.

Solid or liquid media growth of pure culture.  

• For solid media growth, isolated colonies on plated media. 
• For liquid culture, a minimum volume of 4 mL of actively growing culture. 
• Mycobacterium tuberculosis complex and slowly growing Mycobacterium species must show visible growth and be no more than 4 weeks old. 
• Rapidly growing Mycobacterium species must show visible growth and be no more than 10 days old.
• If the culture has inadequate growth, contamination, or mixed Mycobacterium  species, the submitted culture will be rejected and a resubmission will be requested. 

N/A

Ship all cultures at room temperature (DO NOT freeze) for overnight delivery, and prior to Wednesday each week to ensure receipt by Friday.

Shipping of specimens shall be done by a TDG certified individual in accordance with TDG regulations. For additional information regarding classification of specimens for the purposes of shipping, consult either Part 2 Appendix 3 of the TDG Regulations or section 3.6.2 of the IATA Dangerous Goods Regulations as applicable.

None

Notification of sample submission must be emailed or faxed prior to shipping isolates. A requisition for the NRCM must be completed and signed off by the supervisor/designate of the submitting Provincial/Territorial Healthcare laboratory and include the source of specimen, patient gender, date of birth, clinical history, submitting laboratory identifier and submitter information. Please also include isolate characteristics: microscopy, pigmentation, culture characteristics, growth rate/temperature, and other identification. Requested testing for urgent submissions should be accompanied by a justification. The NRCM will provide results to the submitting Provincial/Territorial Healthcare laboratory, who is then responsible for disseminating results to the appropriate local lab or physician.

Cultures will be rejected if appropriate documentation and justification is incomplete or missing

Species identification will be determined primarily through MALDI-TOF. 16S rRNA and/or hsp65 gene fragment sequencing may be used for further differentiation or for confirmation of a submitted identification, as needed.  The identification will be determined by comparison of data to in-house quality controlled databases.  If the isolate is determined to belong to the Mycobacterium tuberculosis complex, it will be automatically referred forMolecular Differentiation of Mycobacterium tuberculosis Complex (MTBC). 

Routine testing from culture: 10 calendar days from the date of specimen receipt.

1. Turenne CY, Tschetter L, Wolfe J, Kabani A. (2001) Necessity of quality-controlled 16S rRNA gene sequence databases: identifying nontuberculous Mycobacterium species. J Clin Microbiol. 39:3637-48.
2. Brunello F, Ligozzi M, Cristelli E, Bonora S, Tortoli E, and Fontana R. (2001) Identification of 54 mycobacterial species by PCR-restriction fragment length polymorphism analysis of the hsp65 gene. J. Clin. Microbiol. 39:2799-2806.