Molecular Detection of Brucella spp. by real-time PCR
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Molecular detection by real-time PCR of Brucella spp.
- Brucellosis
Pure bacterial culture, whole blood – minimum 0.5 mL, cerebrospinal fluid (CSF) – minimum 0.5 mL, biopsy tissue, powder Pure bacterial culture – plate or swab; Whole blood – EDTA tube; Tissue or CSF – submit in a sterile collection tube; powder – submit in a sterile 2 mL o-ring microcentrifuge tube
Pure bacterial culture – plate or swab; Whole blood – EDTA tube; Tissue or CSF – submit in a sterile collection tube; powder – submit in a sterile 2 mL o-ring microcentrifuge tube.
Store blood samples refrigerated until shipped for testing. Store tissue samples and CSF frozen until shipped for testing. Store bacterial cultures refrigerated until shipped for testing.
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.
Suspected Brucella spp. infection.
Completed Bioforensics Assay Development and Diagnostics requisition including sender name, address and telephone number. Patient name or identifier (referring lab #), date of birth, suspected exposure, test(s) requested, clinical history. Type of specimen and date collected. If possible, include the antibiotics administered and travel history.
Real-time PCR is performed using three assays – one specific for Brucella spp., one specific for Brucella abortus, and the third specific for Brucella melitensis. Confirmatory tests of a sample positive for Brucella spp. by real-time PCR result include urease test, susceptibility to Brucella phage Tbilisi, gel formation (Brucella canis), single nucleotide polymorphisms (SNPs), cellular fatty acid (CFA) analysis and omp2a sequencing. CFA analysis and/or 16S sequencing will be performed on all samples negative for Brucella by real-time PCR as long as the sample provides bacterial growth by culture.
4 calendara days for a preliminary result; 14 calendar days for a final result for cultures, and 29 calendar days for a final result for all other sample types.