Development of a high-throughput method for processing sponge-stick samples to detect viable Bacillus anthracis...

Area of Science:

• Environmental microbiology
• Biothreat detection

Background:

• The sponge-stick method for Bacillus anthracis spore detection is limited by low throughput.
• Rapid analysis of numerous samples is crucial after a biological warfare attack.

Purpose of the Study:

• To develop a high-throughput method for simultaneous processing of multiple sponge-stick samples.
• To improve preparedness for rapid response to anthrax incidents.

Main Methods:

• Samples were processed in 50 mL tubes with extraction buffer and shaken.
• Orbital shaking and multi-tube vortexing were employed for spore recovery.
• Simultaneous processing of up to 12 samples was achieved.

Main Results:

• An additional extraction step increased spore recovery by approximately 10%.
• High-throughput method showed comparable spore recovery efficiencies to the traditional method (60-75%).
• The method detected as low as 40 spores per sponge, even with contaminants.

Conclusions:

• The developed high-throughput method significantly enhances the capacity for analyzing Bacillus anthracis spores.
• This method is suitable for integration with Rapid Viability-Polymerase Chain Reaction (RV-PCR) for sensitive detection.
• The improved throughput is critical for effective response and recovery after wide-area anthrax events.