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Biodefense Detection to Protect the Nation

Biodefense Detection to Protect the Nation. Frank Y. S. Chuang & Dora M. Gutierrez Lawrence Livermore National Laboratory J. Kirk Brown Tracy High School. UCRL-PRES-204341. Main Questions. What are germs ? Why are they dangerous ? How do we recognize them?

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Biodefense Detection to Protect the Nation

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  1. BiodefenseDetection to Protect the Nation Frank Y. S. Chuang & Dora M. Gutierrez Lawrence Livermore National Laboratory J. Kirk Brown Tracy High School UCRL-PRES-204341

  2. Main Questions • What are germs? • Why are they dangerous? • How do we recognize them? • How can we protect ourselves against them?

  3. Introduction Throughout history, infectious disease has plagued mankind. Smallpox may have emerged 10,000 years ago in Asia or Africa. Scarred faces of mummies can be found in the Cairo Museum. Smallpox is caused by the variola virus.

  4. Achievements in Infectious Disease Medicine • Vaccines - cause the body to become immune to infectious disease • Antibiotics - kill or stop germs that cause infectious disease Edward Jenner 1749 - 1823 Used cowpox to protect against smallpox. (“Vaccination” from Latin vacca for cow.) Louis Pasteur 1822 - 1895 French chemist who discovered “germ theory of infectious disease” & demonstrated the benefits of sterilization.

  5. So - why worry? New Biological Threats Severe Acute Respiratory Syndrome (SARS) Bacillus anthracis (anthrax) Speed of Epidemic Outbreak

  6. Destructive power of germs • Severe Acute Respiratory Syndrome (SARS) Asia, Canada, U.S. 2003 • AnthraxUnited States, Oct. 2001 • West Nile Virus United States 1999 - ? • Ebola hemorrhagic fever Gabon 2001 • Meningococcal disease Ethiopia 2001 Recent Human Epidemics 20,000,000 18,000,000 16,000,000 14,000,000 12,000,000 10,000,000 8,000,000 6,000,000 4,000,000 2,000,000 70,000 # of deaths Atomic Bomb (Hiroshima) Jewish Holocaust WWII Spanish Flu of 1918

  7. Types of germs

  8. Viral Infection I - Invading host cell

  9. Viral Infection II - Replication of virus

  10. Viral Infection III - Host cell lysis

  11. The Immune System The body’s natural defense against foreign and/or infectious agents: • Skin • White blood cells • Antibodies Antibodies recognize and bind to epitopes of foreign molecules (antigens). Phagocytes “eat” and destroy bacteria.

  12. Immune System: Antibodies • Antibodies help recognize foreign material in the body. • Anything that antibodies stick to will be attacked by the immune system. • Healthy individuals carry trillions of different antibodies in the bloodstream.

  13. Immune System: White Cells T-lymphocytes use antibodies to recognize and destroy infected cells. • White blood cells • macrophages • neutrophils • lymphocytes use either native or adaptive immunity to destroy infectious agents.

  14. War against germs • The immune system works constantly to keep infectious microbes in check. • Germs that can bypass or overwhelm primary defenses cause infectious disease. • Early detection of infectious disease is often the key to preventing widespread outbreak.

  15. Methods of Biodetection Laboratory culture & microscopy Immunoassay (ELISA) DNA hybridization & sequencing

  16. Methods of Biodetection Procedure: put clinical sample in incubator and wait for germs to grow. Look for germs under microscope. Advantage: Laboratory Gold Standard Disadvantage: Results take 7-10 days Laboratory culture & microscopy

  17. How can we make him easier to spot?

  18. Paint him bright purple?? ???

  19. Make himglow-in-the-dark!!

  20. Immunoassays • Antibodies can be used to identify germs • Fluorescent labels allow us to locate antibodies without having to see germs • [Capture antibody layer] + [targeted germs] + [reporter antibody layer] = Sandwich Immunoassay Fluorescent Reporter Antibodies Targeted germs Capture Antibodies

  21. Enzyme-linked Immunosorbent Assay (ELISA) Positive test ELISA is also known as a “sandwich” assay -- because the antibody and antigen layers are stacked like a sandwich.

  22. Methods of Biodetection New methods detect molecules associated with germs, instead of germs themselves. Advantages: very sensitive, results in 1-3 hours, no need for microscope Disadvantages: ? Immunoassay (ELISA) DNA hybridization & sequencing

  23. Advanced Biodetection Need for better technology: • Faster results • More accurate (no false readings) • Higher throughput • Cost-efficient • Automatic

  24. Bead-Based Sandwich Immunoassay Fluorescent reporter molecule binds to microbead only in the presence of targeted antigen. Bead schematics courtesy of Luminex Corp. <http://www.luminexcorp.com/tech/>

  25. Multiplex Optical Encoding

  26. Multiplex Immunoassay 100 bead classes 100 immunoassays influenza adenovirus RSV anthrax etc... Optically encoded beads allow multiple tests to be run on a single sample.

  27. Analyzing the Microbeads

  28. Potential biodefense applications • Hospital-based flow cytometer systems • High-throughput screening of clinical samples • Handheld point-of-care instruments • Local clinic or paramedic “first responder” use • Autonomous monitoring systems • For continuous surveillance in strategic environments

  29. Autonomous Pathogen Detection System (APDS) • Completely autonomous • Sample, concentrate, detect, identify and report. • Multiplex detection • Up to 100 discrete channels. • Detects all bioagent types • Bacterial spore / virus / toxin

  30. Biodefense - A Multidisciplinary Field • Discovery through basic research • Innovative biotechnology • Excellent clinical medicine

  31. Basic Science Research • Basic scientists • biologists • chemists • physicists • discover new mechanisms of disease that could offer new ways to detect or treat infectious disease.

  32. Bioengineering & Biotechnology • Engineers • electrical • mechanical • computer • apply scientific knowledge to develop new medical technologies.

  33. Clinical Medicine Doctors are the first to respond to infectious disease, and also guide the direction of future research and development.

  34. Conclusions • Biodefense is everyone’s business. Public awareness is the first step towards prevention. • “No solution lasts forever.” Science and technology must adapt to cope with the evolving nature of infectious disease. • Future advances in biomedical research depend on experts in all disciplines..!

  35. LLNL M-Division: Medical Physics & Biophysics Candice Cook Christine Paulson Christine P. Nguyen Ben Hindson John T. Chang Steven R. Visuri Mary T. McBride Kodumudi Venkateswaran Bill W. Colston, Jr. UC Davis Medical Center James Carlson, Ph.D. (Clinical Microbiology) Robert Derlet, M.D. (Emergency Medicine) Luminex Corporation Education Outreach - LLNL Marsha McInnis Jason Windsor Acknowledgements

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