Culture

Culture and isolation of Legionella species from clinical or environmental samples associated with clinical illness. Resulting Legionella spp. isolates are subsequently identified and subtyped.

Clinical (respiratory secretions, lung tissue, pleural fluid, extra-pulmonary sites) or environmental samples (water – minimum 1 L, water distribution system swabs in 0.5 – 5 mL of same residual water).

Environmental samples should be collected in sterile containers by methods outlined by Fields (1994) or Joseph et al. (2005).

Samples should be refrigerated, but not frozen, and shipped for testing as soon as possible. Ship on wet ice or cold pack.

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.

Suspected Legionnaires' Disease or Pontiac Fever.

Completed Special Bacteriology requisition form detailing all patient information and relevant clinical information. If possible, attach lab results that have already been done at local or provincial laboratories.

Service done by special request only or to assist in an outbreak. ALL SPECIMENS MUST BE APPROVED by the Special Bacteriology Laboratory before specimens are sent

Samples are pre-treated, by sample type-dependent methods, and seeded to various selective BCYE media. Identification is done by 16S rRNA gene sequencing, mip gene sequencing, and any other relevant tests deemed appropriate for the bacterial identification at the discretion of the Special Bacteriology Laboratory.

50 calendar days. 20 calendar days to confirm negative results. Final report is sent once results from all relevant tests are completed. In cases where staff or resources are limited, final report may be delayed and status of request will be forwarded in a preliminary report.

1. Fields, B. S. 1994. Procedures for the Recovery of Legionella from the Environment. Public Health Service, Centers for Disease Control and Prevention (CDC), Atlanta GA.
2. Fry, N. K., S. Warwick, N. A. Saunders, and T. M. Embley. 1991. May. The use of 16S ribosomal RNA analyses to investigate the phylogeny of the family Legionellaceae. J. Gen. Microbiol. 137:1215-1222
3. Isenberg, Henry D., Clinical Microbiology Procedures Handbook, "Direct Immunofluorescent-Antibody Examination for Legionella spp.," Section 9.5, American Society for Microbiology, 1992
4. Joseph, C., J. Lee, J. van Wijngaarden, V. Drasar, and M. Castellani-Pastoris. 2005. European Guidelines for Control and Prevention of Travel Associated Legionnaires' Disease In Anonymous. European Working Group for Legionella Infections.
5. Ratcliff, R. M., J. A. Lanser, P. A. Manning, and M. W. Heuzenroeder. 1998. Jun. Sequence-based classification scheme for the genus Legionella targeting the mip gene. J. Clin. Microbiol. 36:1560-1567
6. Wilkinson, H. W. 1987. Hospital-Laboratory Diagnosis of Legionella infections. Centers for Disease Control, Atlanta, GA.