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Monitoring and Detecting Anthrax By Jennifer Bisogno Eastern Connecticut State Univ.
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Analytical Chemistry A Minisonicator to Rapidly Disrupt Bacterial Spores for DNA Analysis Belgrader, Hansford, Kovacs, Venkateswaran, Mariella, Milanovich, Nasarabadi, Okuzumi, Pourahmadi, Northrup
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Anthrax Top candidate for biological weapon Spores produced in large quantities Stored for decades readily disseminated in air
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Defense against Anthrax Instrumentation that provides early warning identifying infected individuals quickly administer antibiotics high specificity to distinguish virulent strains
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Minisonicator Improves PCR analysis and detection Decreases limit of detection Reduces time of detection Increases signal amplitude
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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
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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
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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
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Figure 1
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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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