Molecular Detection of Bacillus anthracis

Molecular detection by real-time PCR of Bacillus anthracis.

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 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 Bacillus anthracis 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 in-house differentiating molecular assays specific for the pXO1 and pXO2 regions of Bacillus anthracis, as well as a Bacillus anthracis chromosomal target. Confirmatory tests of a sample positive for Bacillus anthracis by real-time PCR result include gamma phage lysis, motility, penicillin sensitivity, Multiple Locus Variable Number Tandem Repeat Analysis (MLVA), cellular fatty acid (CFA) analysis, and 16S sequencing. CFA analysis and/or 16S sequencing will be performed on all samples negative for Bacillus anthracis by real-time PCR, as long as the sample provides bacterial growth by culture.

4 calendar days for a preliminary result and 14 calendar days for a final result.