Molecular detection by PCR From Clinical Material
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Molecular detection of Legionella pneumophila by PCR from clinical material.
- Legionnaires' disease
- Pontiac fever
Bronchoalveolar lavage (BAL), nasopharyngeal aspirate (NPA), throat swab, sputum, cerebrospinal fluid (CSF), and tissue samples. Dry swabs will not be accepted for testing; any swabs sent must be supplied in appropriate storage medium. At least 1.0 mL of fluid sample is required for testing, and it is preferable if all samples are supplied in screw-capped tubes made of freeze-thaw and shatter-resistant plastic.
Please follow standard aseptic sampling methods for collecting specimens and ensure any tissue samples/swabs are in appropriate storage medium.
No further processing at the sending lab is required once specimens are collected according to the above instructions. It is preferable that all specimens be frozen after collection. If that’s not possible, then store at refrigeration temperature (2 – 8°C). Ensure that specimens are held at the appropriate temperature when shipped to the NML (e.g. frozen specimens shipped with dry ice, refrigerated specimens with cold packs).
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 (most recent version of requisition is required), detailing all patient information and relevant clinical information. If possible, attach lab results that have already been done at local or provincial laboratories.
If a test performed by the submitting lab produced a positive result for L. pneumophila, it is also helpful to indicate this in the documentation including what type of test was performed.
DNA is extracted using a commercially available kit and results are based upon a real-time PCR assay targeting the mip gene region specific for L. pneumophila. Positive samples are confirmed using a secondary real-time PCR assay targeting the ssrA gene specific to Legionella species.
10 calendar days. Please note that during times when large numbers of samples are received or if tests must be repeated the turnaround time may be longer.
- Benitez A.J., Winchell, J. M. 2012. Clinical application of a multiplex real-time PCR assay for simultaneous detection of Legionella species, Legionella pneumophila, and Legionella pneumophila serogroup 1. J. Clin. Microbiol. 51 (1): 348-351.