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Detection of Bartonella species by PCR and sequencing

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Description:

Detection of Bartonella species by PCR and sequencing.

 

Test Category:
Molecular Detection and Genotyping
Pathogen:
Bartonella spp.
Laboratory:
Illnesses and Diseases:
  • Bartonellosis (Trench Fever, Cat-scratch disease)
Specimen:

Fluid aspirate from wound, fluid and / or tissue from lymph nodes, heart valve biopsy, paraffin block, synovial fluid or whole blood Minimum required volumes are as follows:

  • Liquid specimens including whole blood - 0.5 mL
  • Fresh tissue (skin biopsy) – 1 to 5 mg

CSF samples are accepted for testing however, they are not ideal specimen types as they are typically low-yield.

Collection Method:

Collect blood in EDTA tubes. AVOID HEPARIN. Do not centrifuge. Collect a swab, fluid aspirate or tissue biopsy from suspected area of infection. Swabs must be supplied in at least 1 mL appropriate storage medium and NOT shipped dry.  Aseptically collect samples into sterile, leak-proof containers made of freeze-thaw and shatter-resistant plastic.

Specimen Processing, Storage and Shipping:

Store specimens refrigerated for up to 5 days and ship with freezer packs. If > 5 days, store at -20 °C and ship on dry ice. Paraffin-blocks can be stored and shipped at ambient temperature.

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:

Clinical manifestations of B. henselae infection, also referred to as “Cat-scratch disease” (CSD), may include localized infections such as subacute regional lymphadenopathy, skin lesions, musculoskeletal lesions, culture-negative endocarditis, and uveitis and retinitis; however systemic manifestations are also common resulting in symptoms of fever, meningitis, osteomyelitis and arthritis.

Infection with B. quintana (Trench fever) is characterized by fever, which may be present as a single bout or bouts of recurrent fever, headache, rash and bone pain mainly in the shins neck and back.

Both B. henselae and B. quintana may cause bacillary angiomatosis in immunocompromised individuals such as those with advanced HIV. 

Accompanying Documentation:

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

 

Comments:

Due to the fact that a Bartonella spp. infection can result in localized and systemic manifestations, it is difficult to accurately detect the infectious agent unless the appropriate specimens are submitted for testing. Please only submit the listed specimen types. Samples may be subject to rejection if they are not the appropriate specimen type, have insufficient volume or are not accompanied by relevant patient information or clinical history.

 

Methods and Interpretation of Results:

Extracted DNA is initially tested by a real-time PCR assay capable of detecting all Bartonella species. Samples positive by the screening assay, are tested by additional real-time PCR assays specific for B. henselae and B. quintana. If necessary, conventional PCR and sequencing for species determination is performed.   

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. Raoult D, Roblot F, Rolain JM, Besnier JM, et al. 2006. First isolation of Bartonella alsatica from a valve of a patient with endocarditis. J Clin Microbiol. 44:278–9.
  2. Angelakis, E., Rolain, J. M., Raoult, D., & Brouqui, P. 2011. Bartonella quintana in head louse nits. Pathogens and Disease, 62(2), 244-246.
  3. Chomel B, Boulouis HJ, Maruyama S, Breitschwerdt EB. 2006. Bartonella spp. in pets and effect on human health. Emerg. Infect. Dis. (12):389-394.
  4. Raoult D. 2007. From cat scratch disease to Bartonella henselae infection. Clin. Infect. Dis. (45):1541-1542.
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