Flow-cytometric Analysis of Bacillus anthracis Spores

Abstract

Flow-cytometric technique has been established as a powerful tool for detection and identification of microbiological agents. Unambiguous and rapid detection of Bacillus anthracis spores has been reported using immunoglobulin G-fluorescein isothiocyanate conjugate against live spores. In addition to the high sensitivity, the present technique could differentiate between spores of closely related species, eg, Bacillus cereus and Bacillus subtilis using fluorescence intensity. The technique can be used for detection of live as well as inactivated spores making it more congenial for screening of suspected samples of bioterrorism.

Real-time loop-mediated isothermal amplification assay for rapid and sensitive detection of anthrax spores in spiked soil and talcum powder

“…Spores were scraped using Phosphate Buffer Saline (PBSpH 7.2) from incubated plates and washed three times with triple distilled water at 3,000g for 20 min followed by heat shock treatment at 60°C for 90 min; spores were centrifuged at 3,000g for 20 min to remove cellular debris. The spore pellet obtained was washed twice with triple distilled water at 3,000g for 20 min (Kamboj et al 2006). Finally the spore pellet was suspended in phosphate buffer saline (PBS) and its concentration was determined by direct colony counting on Trypticase Soy Agar (TSA; Difco, Sparks, MD, USA).…”

Identification and Characterization of Bacillus anthracis Spores by Multiparameter Flow Cytometry

“…Comparatively, spore-based methods that analyze intact spores can be rapid because surface antigens are detected directly by using simple labeling procedures (5); however, many of these strategies are not selective for specific members of the B. cereus group (9,17). Through a phage display screening process, short peptide fragments that exhibited species-specific binding to Bacillus spores were discovered (19,21), and several investigators have used them successfully (2,15).…”