Presentation on theme: "Monitoring and Detecting Anthrax By Jennifer Bisogno Eastern Connecticut State Univ."— Presentation transcript:
Monitoring and Detecting Anthrax By Jennifer Bisogno Eastern Connecticut State Univ.
Analytical Chemistry A Minisonicator to Rapidly Disrupt Bacterial Spores for DNA Analysis Belgrader, Hansford, Kovacs, Venkateswaran, Mariella, Milanovich, Nasarabadi, Okuzumi, Pourahmadi, Northrup
Anthrax Top candidate for biological weapon Spores produced in large quantities Stored for decades readily disseminated in air
Defense against Anthrax Instrumentation that provides early warning identifying infected individuals quickly administer antibiotics high specificity to distinguish virulent strains
Minisonicator Improves PCR analysis and detection Decreases limit of detection Reduces time of detection Increases signal amplitude
Bacterial Strain and Spore Purification Resuspention and quantitation by spreading on agar plates containing BHI incubation in shaking water bath for 3 days sporulation checked periodically under phase contrast microscope centrifuged, washed,and resuspended Lysozome added and reincubated sonication
Spore Lysis by Sonication 5mg of glass beads and 75-100uL B. Anthracis spores placed in tube tube placed in sonicating water bath beads provide more surface area high pressure and temperature will damage cells
Results Anylized by quantitative real-time PCR using the ABI Prism 7700 spectrofluorometric thermo cycler measures relative level of available DNA improvement in PCR detection as result of sonication reduced detection limit
Conclusion Sonication to purified spores improved limit of detection essential step in rapid detection small portable devices wanted goal to build a portable spore lysis cartridge with a minisonictor
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