Molecular Detection of Yersinia pestis by real-time PCR

Molecular detection by real-time PCR of Yersinia pestis.

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 Yersinia pestis 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 for 3 targets specific to Yersinia pestis. Confirmatory tests of a sample positive for Yersinia pestis by real-time PCR result include direct fluorescent antibody (DFA) staining, bacteriophage lysis, cellular fatty acid (CFA) analysis, and 16S sequencing. CFA analysis and/or 16S sequencing will be performed on all samples negative for Yersinia pestis 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.