Identification and Subtyping from Pure Cultures

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Requisition Forms

Reference Details

Description:

Identification of Legionella pneumophila and subtyping it by Sequence-based typing (SBT) for Serogroups 1-14. If isolate is a L. pneumophila Serogroup 1, then further subtyping by a monoclonal antibody (mAb) panel is done.

Test Category:
Genotyping
Pathogen:
Legionella pneumophila
Illnesses and Diseases:
  • Legionnaires' disease
Specimen:

Pure culture.

Collection Method:

BCYE plates, slants, or charcoal transport swabs are acceptable. Isolated colonies on BCYE plate are preferred.

Specimen Processing, Storage and Shipping:

Store samples between room temperature and 37oC until shipped for testing. Ship at room temperature.

Transportation of Dangerous Goods:

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.

For additional guidance on the transport of infectious substances in other languages, please click on the link below.

http://www.who.int/ihr/capacity-strengthening/infectious-substances/en/

Patient Criteria:

Suspected Legionnaires' Disease or Pontiac Fever.

Accompanying Documentation:

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.

 

Comments:

All patient and strain history must be included. 

Methods and Interpretation of Results:

Legionella pneumophila is identified and subtyped using SBT. This involves the amplification and sequencing of seven target genes, including some under diversifying pressure, for the purpose of strain typing.  It is an adaptation of multi-locus sequence typing (MLST), where all gene targets are non-selective (eg. house-keeping genes).  Furthermore, the serogroup is determined by latex agglutination, and if required, direct fluorescent assays (DFA), using ProLab Diagnostics Serogroups 1-14 antibody kits. L. pneumophila Serogroup 1 is further typed using the Dresden Legionella LPS mAb subtyping panel.

Turnaround Time:

20 calendar days.  In cases where staff or resources are limited final report may be delayed and status of request will be forwarded in a preliminary report.  Final report is sent once all results are complete.

 

Contact:
Phone: (204) 789-2137
Fax: (204) 784-7509
References:
  1. Farhat, C., M. Mentasti, E. Jacobs, N. K. Fry, and C. Lück. 2011. Dec. The N-Acylneuraminate Cytidyltransferase Gene, neuA, Is Heterogenous in Legionella pneumophila Strains but Can Be Used as a Marker for Epidemiological Typing in the Consensus Sequence-Based Typing Scheme. J. Clin. Microbiol. 49:4052-4058
  2. Gaia, V., N. K. Fry, B. Afshar, P. C. Luck, H. Meugnier, J. Etienne, R. Peduzzi, and T. G. Harrison. 2005. May. Consensus sequence-based scheme for epidemiological typing of clinical and environmental isolates of Legionella pneumophila. J. Clin. Microbiol. 43:2047-2052
  3. Gilmour, M. W., K. Bernard, D. M. Tracz, A. B. Olson, C. R. Corbett, T. Burdz, B. Ng, D. Wiebe, G. Broukhanski, P. Boleszczuk, P. Tang, F. Jamieson, G. Van Domselaar, F. A. Plummer, and J. D. Berry. 2007. Mar. Molecular typing of a Legionella pneumophila outbreak in Ontario, Canada. J. Med. Microbiol. 56:336-341
  4. Helbig, J. H., S. Bernander, M. Castellani Pastoris, J. Etienne, V. Gaia, S. Lauwers, D. Lindsay, P. C. Luck, T. Marques, S. Mentula, M. F.   Peeters, C. Pelaz, M. Struelens, S. A. Uldum, G. Wewalka, and T. G. Harrison. 2002. 10. Pan-European study on culture-proven Legionnaires' disease: distribution of Legionella pneumophila serogroups and monoclonal subgroups. Eur. J. Clin. Microbiol. Infect. Dis. 21:710-716
  5. Legionella pneumophila Sequence-Based Typing Database website (http://bioinformatics.phe.org.uk/legionella/legionella_sbt/php/sbt_homepage.php) developed by The European Working Group for Legionella Infections (EWGLI).
  6. Ratzow, S., V. Gaia, J. H. Helbig, N. K. Fry, and P. C. Luck. 2007. Jun. Addition of neuA, the gene encoding N-acylneuraminate cytidylyl transferase, increases the discriminatory ability of the consensus sequence-based scheme for typing Legionella pneumophila serogroup 1 strains. J. Clin. Microbiol. 45:1965-1968
  7. Wilkinson, H. W. 1987. Hospital-Laboratory Diagnosis of Legionella infections. Centers for Disease Control, Atlanta, GA.
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