Molecular Detection of Brucella spp. by real-time PCR

Molecular detection by real-time PCR of classical brucellosis causing 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.

Store bacterial cultures refrigerated until shipped for testing.  Store blood samples refrigerated until shipped for testing.  Store tissue samples and CSF frozen 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 brucellosis causing 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.

Please notify the laboratory prior to submission of specimen.

Real-time PCR is performed using three assays – one specific for classic brucellosis causing Brucella spp., one specific for Brucella abortus, and one specific for Brucella melitensis.

Confirmatory tests of a sample positive for Brucella spp. by real-time PCR result may include urease test, susceptibility to Brucella phage Tbilisi, gel formation (Brucella canis), Single Nucleotide Polymorphisms (SNPs), Whole Genome Sequencing (WGS), and Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) bacterial analysis,

MALDI-TOF MS will be performed on all samples negative for Brucella by real-time PCR, as long as the sample provides bacterial growth by culture.

4 calendar days for a preliminary result; 14 calendar days for a final result for standard sample type; 35 calendar days for a final result for any non-standard specimen type, specialized testing requests or  clinical samples such as blood or tissue requiring a 28 day incubation period.

1. Ferreira, L, et al. 2010.  Identification of Brucella by MALDI-TOF mass spectrometry.  Fast and reliable identification from agar plates and blood cultures.  PLOS One 5, e14235.
2. Tracz, DM, et al. 2012.  Effect of gamma radiation on the identification of bacterial pathogens by MALDI-TOF MS. J. Microbiol. Methods 92, 132-134.

Fact Sheet - Brucellosis

Brucella spp. (B. abortus, B. canis, B. melitensis, B. suis) - Material Safety Data Sheets (MSDS)

Brucella and Ochrobactrum Taxonomic Updates for Laboratories | ASM.org

National case definition: Brucellosis