Detection of Leptospira species by PCR and sequencing
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Reference Details
Molecular detection of Leptospira species in clinical specimens.
- Leptospirosis
Whole blood, urine or cerebrospinal fluid (CSF). Minimum required volumes are as follows:
- Whole Blood – 5 mL
- Urine (fresh) – 5 mL
- CSF – 0.5 mL
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.
Store specimens refrigerated up to 5 days and ship with freezer packs. If > 5 days, freeze at -20 °C and ship on dry ice.
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.
Symptoms of Leptospira spp. infection may include fever, headache, chills, severe malaise, skin rash, vomiting, diarrhea, 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.
Completed Requisition for Molecular Testing for Selected Zoonotic Agents.
Specimens may be subject to rejection if they are not the appropriate sample type, have insufficient volume or not accompanied by relevant patient information or clinical history. This test is performed for investigational purposes.
Extracted DNA is screened by an in-house real-time PCR assay specific for Leptospira spp. If positive by real-time PCR, samples are subject to confirmation testing by real-time PCR or conventional PCR. Initiation of antibiotic treatment prior to testing may result in decreased bacterial genome which will affect the outcome of PCR testing.
21 calendar days.
- Levett, P.N. Leptospirosis (2001) Clin. Micro. Rev. (14): 296-326.
- Stoddard R. A., Gee J. E., Wilkins P. P., McCaustland K., et al.2009. Detection of pathogenic Leptospira spp. through TaqMan polymerase chain reaction targeting the LipL32 gene. Diagn. Microbiol. Infect. Dis. 64:247–255.