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Monitoring and Detecting Anthrax By Jennifer Bisogno Eastern Connecticut State Univ.

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Presentation on theme: "Monitoring and Detecting Anthrax By Jennifer Bisogno Eastern Connecticut State Univ."— Presentation transcript:

1 Monitoring and Detecting Anthrax By Jennifer Bisogno Eastern Connecticut State Univ.

2 Analytical Chemistry A Minisonicator to Rapidly Disrupt Bacterial Spores for DNA Analysis Belgrader, Hansford, Kovacs, Venkateswaran, Mariella, Milanovich, Nasarabadi, Okuzumi, Pourahmadi, Northrup

3 Anthrax Top candidate for biological weapon Spores produced in large quantities Stored for decades readily disseminated in air

4 Defense against Anthrax Instrumentation that provides early warning identifying infected individuals quickly administer antibiotics high specificity to distinguish virulent strains

5 Minisonicator Improves PCR analysis and detection Decreases limit of detection Reduces time of detection Increases signal amplitude

6 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

7 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

8 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

9 Figure 1

10 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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