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Waterborne Pathogen Prevention and Detection Using Traditional Methods and Microarray Probe Detection

Waterborne Pathogen Prevention and Detection Using Traditional Methods and Microarray Probe Detection. Claribel Orellana CE 421 12/5/07. Biotechnology. Priorities: Agriculture and Medicine Malaria Vaccine Stem cell research Completion of the human genome map

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Waterborne Pathogen Prevention and Detection Using Traditional Methods and Microarray Probe Detection

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  1. Waterborne Pathogen Prevention and Detection Using Traditional Methods and Microarray Probe Detection Claribel Orellana CE 421 12/5/07

  2. Biotechnology • Priorities: Agriculture and Medicine • Malaria Vaccine • Stem cell research • Completion of the human genome map • ‘Golden Rice’ modified to make vitamin A • GCSF for increasing white blood count in chemotherapy patients

  3. World Concern: Water Quality • Half the world affected by contaminated water • Unsanitary conditions and lack of resources • Cause: waterborne pathogens

  4. Waterborne Pathogens • Three different types: bacteria, viruses, and bacteria

  5. Typical Contamination • Unprotected water source • Inadequate sanitation • Animal and fecal matter reaching water source • Surface runoff through the ground, water pipes and wells Well Cattle pond Well Surface drain Cattle pond

  6. Typical Contamination • Animal and fecal matter reaching water source • Surface runoff through the ground, water pipes and wells

  7. Traditional Practices for Safe Water • Point-of-use disinfection • Use of sodium and calcium hypochlorite • Safer and easier to use and distribute • Destroys most pathogens • Electrolysis • System run on solar power • Generators can generate enough disinfectant for 10,000 people. • Inexpensive

  8. Traditional Practices for Safe Water • Safe Storage • Stored water vs. municipal tap • Container comparison • CDC container • Cantero

  9. DNA Microarrays • DNA microarrays: reverse dot-blots for which sequence-specific “probes” are attached to substrate in a lattice pattern • spots are usually 100-200 micrometers and 200-500 micrometers away from each other and they represent specific probe sequences • Simultaneous detection vs. cultivation

  10. Polymerase Chain Reaction (PRC) • PCR amplifies DNA sequences

  11. Microarray Process • Sequences are hybridized • Specific bacterial targets are detected

  12. Precision Factors • Assay sensitivity • Sample size • Efficiency of pathogen isolation • Efficiency of nucleic acid extraction • Effect of co-precipitating factors that inhibit PCR

  13. Benefits and Limitations • Benefits: • Simultaneously detects pathogens • Not limited to identification by product length • Limitations: • Requires pathogens to be identified before configuring array • Needs to be validated • Currently more expensive than traditional methods

  14. Microarray Flow Process

  15. Application • EPA looking into feasibility • Is being monitored • Will most likely become a standard • Soon be more cost-effective • Priority is quick pathogenic detection

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