Molecular detection of Burkholderia mallei/pseudomallei by real-time PCR

Molecular detection by real-time PCR of Burkholderia mallei/pseodomallei.

• Melioidosis

Pure bacterial culture, whole blood – minimum 0.5 mL, cerebrospinal fluid (CSF) – minimum 0.5 mL, fresh 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 Burkholderia pseudomallei/mallei 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, travel history, type of specimen and date collected. 

Please notify the laboratory prior to submission of specimen.

Real-time PCR is performed using in-house differentiating molecular assays which distinguish between Burkholderia pseudomallei and Burkholderia mallei.

Confirmatory tests of a sample positive for B. pseudomallei/B. mallei by real-time PCR result may include gentamicin sensitivity, oxidase production, motility, sucrose/maltose utilization and Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) bacterial identification.

MALDI-TOF MS will be performed on all samples negative for B. pseudomallei/B. mallei 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; 28 calendar days for any non-standard specimen type or specialized testing requests

1. 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.

Burkholderia (Pseudomonas) pseudomallei - Material Safety Data Sheets (MSDS)