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Detection of Leptospira species by PCR and sequencing

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Reference Details

Description:

Detection of Leptospira spp. by PCR and sequencing.

Test Category:
Molecular Detection and Genotyping
Pathogen:
Leptospira spp.
Laboratory:
Illnesses and Diseases:
  • Leptospirosis
Specimen:

Whole blood, urine or CSF. Minimum required volumes are as follows:

  • Whole Blood –  2 mL
  • Urine (fresh) 5 mL
  • CSF – 0.5 mL

 

Collection Method:

Collect blood in EDTA tubes. AVOID HEPARIN. Do not centrifuge. Aseptically collect urine or CSF into sterile, leak-proof container made of freeze-thaw and shatter-resistant plastic, without additives. 

Specimen Processing, Storage and Shipping:

Store specimens refrigerated up to 5 days and ship with freezer packs. If > 5 days, freeze at -20 °C and ship on dry ice.

 

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.

 

Patient Criteria:

Symptoms of Leptospira spp. infection may include fever, headache, chills, severe malaise, skin rash, vomiting, diarrhoea, myalgia, conjunctival suffusion, jaundice, kidney and/or liver failure, meningitis and chest pains. Contact with an infected animal (dogs, cattle, raccoons, rodents, etc.) or recent travel to locations known to be endemic for Leptospira spp.

Accompanying Documentation:

Completed Requisition for Molecular Testing for Selected Zoonotic Agents.

 

Comments:

Specimens may be subject to rejection if they are not the appropriate sample type, have insufficient volume or are not accompanied by relevant patient information or clinical history.

Initiation of antibiotic treatment prior to testing may result in decreased antibody production which will affect the outcome of serological testing.

THIS TEST IS PERFORMED FOR INVESTIGATIONAL OR RESEARCH PURPOSES ONLY

Methods and Interpretation of Results:

Extracted DNA is screened by real-time PCR using an assay that targets the lipL32 gene of pathogenic Leptospira spp. Samples that are positive by the screening assay are then tested by a confirmatory real-time PCR assay targeting Leptospira 16S rRNA and conventional PCR and sequencing of the citrate synthase gene.

 

Turnaround Time:

21 calendar days.

Contact:
Phone #: (204) 789-7434 or (204) 789-6060
Fax: (204) 789-2082
References:
  1. Levett, P.N. Leptospirosis (2001) Clin. Micro. Rev. (14): 296-326.
  2. Stoddard R. A., Gee J. E., Wilkins P. P., McCaustland K., et al.2009Detection of pathogenic Leptospira spp. through TaqMan polymerase chain reaction targeting the LipL32 gene. Diagn. Microbiol. Infect. Dis. 64:247255.
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