Mycobacterial Interspersed Repetitive-Unit Variable Number of Tandem Repeats (MIRU-VNTR) genotyping

Primary typing of Mycobacterium tuberculosis complex (MTBC) isolates using Variable Number of Tandem Repeat analysis based molecular typing to determine relatedness of strains.

Solid or liquid media growth.

• For solid media growth, isolated colonies on plated media are preferred. Mycobacterium tuberculosis complex species must show visible growth and be no more than 4 weeks old.

• For liquid culture, a minimum volume of 4 mL of actively growing culture is preferred.

• If there is inadequate growth, or there is contamination, submitted cultures 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

Requisitions must be faxed prior to shipping isolates to NRCM - attention Joyce Wolfe, Program Chief, NRCM/NML at 204-789-2036. Requisition for the NRCM must be completed and signed off by the supervisor/designate of the submitting laboratory and include the source of specimen, patient gender, date of birth, clinical history, submitting laboratory identifier and submitter information. Also include isolate characteristics including microscopy and growth rate if relevant. Requested testing for urgent submissions should be accompanied with justification.

Cultures will be rejected if appropriate documentation is incomplete or missing.

MIRU-VNTR patterns will be determined as per the "Multilocus Variable Number Tandem Repeat Genotyping of Mycobacterium tuberculosis Technical Guide". The 24 locus MIRU-VNTR patterns will be provided with a dendrogram including other closely related strains from the requesting province.

Urgent/stat submissions: 7 calendar days from the date of specimen receipt. Routine testing from culture: 30 calendar days from the date of specimen receipt. Submissions of stock culture: 44 calendar days from the date of specimen receipt.

1. Technical guidelines can be accessed in the Multilocus Variable Number Tandem Repeat Genotyping of Mycobacterium tuberculosis Technical Guide available at the following website: http://www.miru-vntrplus.org/MIRU/files/MIRU-VNTRtypingmanualv6.pdf

2. Supply P, Lesjean S, Savine E, Kremer K, van Soolingen D, and Locht C. (2001). Automated high-throughput genotyping for study of global epidemiology of Mycobacterium tuberculosis based on mycobacterial interspersed repetitive units. J. Clin. Microbiol. 39:3563-3571.

3. Blackwood KS, Wolfe JN, Kabani AM. (2004) Application of mycobacterial interspersed repetitive unit typing to Manitoba tuberculosis cases: can restriction fragment length polymorphism be forgotten? J Clin Microbiol. 42:5001-6.

4. de Beer J, Kremer K, Kodmon C, Supply P, van Soolingen. (2011). First Worldwide Proficiency study on Variable-Number Tandem Repeat Typing of Mycobacterium tuberculosis complex strains. J Clin Microbiol. 50: 662-669.

5. Christianson S, Wolfe J, Orr P, Karlowsky J, Levett PN, Horsman GB, Thibert L, Tang P, Sharma MK. Evaluation of 24 locus MIRU-VNTR genotyping of Mycobacterium tuberculosis isolates in Canada. 2010. Tuberculosis 90:31-38