Molecular Detection by PCR and Sequencing
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Molecular detection of Borrelia burgdorferi sensu lato
- European Lyme Disease
- Neuroborreliosis
- North American Lyme Disease
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.
- 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.
Store specimens refrigerated up to 5 days and ship with freezer packs. If > 5 days, store 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.
Appropriate clinical symptoms with potential exposure to Ixodes spp. 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.
Completed Requisition for Molecular Testing for Selected Zoonotic. If possible, include the clinical history and lab results performed at local or provincial laboratories.
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 purposes.
Extracted DNA is screened by a real-time PCR assay specific for Borrelia species. If positive by real-time PCR, samples are subject to confirmatory testing by real-time PCR, melt-curve analysis or conventional PCR for species confirmation.
Initiation of antibiotic treatment prior to testing may result in decreased bacterial genome which will affect the outcome of PCR testing.
21 calendar days.
- 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.
- 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.
- Scott, J.C., Wright, D.J. M., and Cutler, S.J. 2005. Typing African Relapsing Fever Spirochetes. Emerg. Inf. Dis. 11(11):1722-1729.
- 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 Diseases, 16(5), 556–564.