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Molecular Detection by PCR and Sequencing

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Requisition Forms

Reference Details

Description:

Molecular detection of Borrelia species including, but not limited to; B. burgdorferi sensustricto, B. afzelii, B. garinii, B. mayonii and B. spielmanii. in clinical specimens.

Test Category:
Molecular Detection and Genotyping
Pathogen:
Borrelia burgdorferi sensu lato
Laboratory:
Illnesses and Diseases:
  • European Lyme Disease
  • Neuroborreliosis
Specimen:

Synovial fluid, cerebrospinal fluid (CSF) or tissue biopsy. Minimum required volumes are as follows:

  • Synovial fluid – 0.5 mL
  • CSF - 0.5 mL.
  •  Fresh tissue (skin biopsy) – 1 to 5 mg

Serum is a low-yield specimen type and will be rejected.

Collection Method:
  • Aseptically collect CSF or synovial fluid into sterile leak-proof container without additives.
  • Fresh or paraffin-embedded tissue:
    • Wash biopsy site with an antiseptic soap. Thoroughly rinse area with sterile water. Do not use alcohol or iodine preparations. A local anaesthetic may be used.
    • Collect punch biopsy to include full thickness of dermis. Place tissue in a saline-moistened gauze and transfer into a sterile, leak-proof container containing 2 mL saline.  
Specimen Processing, Storage and Shipping:

Store specimens refrigerated up to 5 days and ship with freezer packs. If > 5 days, store 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:

Appropriate clinical symptoms with potential exposure to blacklegged ticks. Typical symptoms include fever, headache, fatigue, and a characteristic rash may be present. If left untreated, infection can spread to joints, the heart, and the nervous system.

Accompanying Documentation:

Completed Requisition for Molecular Testing for Selected Zoonotic.  If possible, include the clinical history and lab results performed at local or provincial laboratories.

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.

THIS TEST IS PERFORMED FOR INVESTIGATIONAL OR RESEARCH PURPOSES ONLY
 

Methods and Interpretation of Results:

Extracted DNA is screened by a real-time PCR assay specific for Borrelia species. If positive by real-time PCR, samples are tested by melt-curve analysis, conventional PCR and sequencing for confirmation and species determination.

Initiation of antibiotic treatment prior to testing may result in decreased bacterial genome which will affect the outcome of PCR testing.

Turnaround Time:

21 calendar days.

Contact:
Phone #: (204) 789-7434 or (204) 789-6060
Fax: (204) 789-2082
References:
  1. Courtney, J.W., Kostelnik, L.M., Zeidner, N.S., Massung, R.F. 2004.  Multiplex real-time PCR for detection of Anaplasma phagocytophilum and Borrelia burgdorferi.  J. Clin. Micro. 42(7):3164-3168.
  2. Johnson, B.J.B., Happ, C.M., Mayer, L.W., Piesman, J.    1992.  Detection of Borrelia burgdorferi in ticks by species-specific amplification of the flagellin gene. Am. J. Trop. Med. Hyg., 47(6):730-741.
  3. Scott, J.C., Wright, D.J. M., and Cutler, S.J. 2005. Typing African Relapsing Fever Spirochetes. Emerg. Inf. Dis. 11(11):1722-1729.
  4. Pritt, B. S., Mead, P. S., Hoang Johnson, D. K., Neitzel, et. al. 2016. Identification of a novel pathogenic Borrelia species causing Lyme borreliosis with unusually high spirochaetaemia: a descriptive study. The Lancet. Infectious Diseases16(5), 556–564.
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