Molecular Detection

Molecular detection of Anaplasma phagocytophilum in clinical specimens.

Whole blood or buffy coat specimens are recommended. Cerebrospinal fluid (CSF) from patients whose clinical conditions suggests meningitis or serum samples are acceptable.  Minimum required volumes are as follows:

Whole blood – 5.0 mL

Buffy Coat – 0.5 mL

CSF – 0.5 mL

Serum – 0.5 mL

Collect blood in EDTA tubes. Avoid heparin. Do not centrifuge.  Aseptically collect buffy coat, CSF or serum into sterile, leak-proof containers 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, 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 within the previous 5-21 days prior to symptom onset . Clinical evidence of anaplasmosis is characterized by an acute onset of fever with one or more of the following non-specific symptoms: chills, malaise, headache, myalgia, arthralgia, leukopenia, thrombocytopenia or elevated hepatic transaminases.

Completed Requisition for Molecular Testing for Selected Zoonotic Agents. 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 not accompanied by relevant patient information. This test is performed for investigational purposes.

Testing is performed, in whole or in part, using a lab-developed test which has not been fully validated/verified.

Extracted DNA is screened by an in-house real-time PCR assay specific for A. phagocytophilum.  If positive by real-time PCR, samples are subject to confirmation testing by real-time PCR and may be tested by conventional PCR and sequencing for strain determination.

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

21 calendar days.

1. Massung, R.F., Owens, J.H., Ross, D., et al. 2000. Sequence analysis of the ank gene of granulocytic ehrlichiae. J. Clin. Microbiol. 38(8):2917-2922.
2. 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.
3. Krakowetz CN, Dibernardo A, Lindsay LR, Chilton NB. 2014. Two Anaplasma phagocytophilum Strains in Ixodes      scapularis Ticks, Canada. Emerg. Inf. Dis. 20(12):2064-2067.