Dr. Jeri Veenstra, Lee University

Report from Chautauqua Short Course #76

July 18 20, 2004

Clark Atlanta University

Dr. Suzanne Barth, University of Texas, Austin

Epidemiology: Molecular Methods for Subtyping Bacterial Microorganisms

 

My purpose in attending this course was to supplement my teaching of  Epidemiology of Infectious Disease.  The sessions provided a good overview of emerging and re-emerging infectious diseases, including bacterial, viral, fungal, protozoal and prion-caused diseases.  Currently used molecular diagnostics were presented and Dr. Barth provided many practical examples of how these techniques were used in the Texas Department of Health for detecting and tracking outbreaks of infectious disease.  

 

We compared conventional and molecular subtyping methods of epidemiologic investigation.  Dr. Barth holds such a wealth of experience in this field that she was able to guide us through several scenarios in which both methods are invaluable tools in the hands of the investigative team.  We examined amplification, assay and probe techniques for use in genotyping.  We reviewed techniques involving pulse-field gel electrophoresis, multilocus enzyme electrophoresis, restriction fragment length polymorphism and restriction endonuclease analysis.

 

We looked specifically at outbreaks involving food contamination as this is an area of special interest for Dr. Barth.  We were fortunate to be able to tour the Centers for Disease Control in Atlanta on our third day.  We visited the labs involved in Salmonella and Shigella research and molecular subtyping.  We enjoyed seeing how the techniques we had just studied were actually being used in the working labs.  We had fun in the visitor's center where we learned the story of the CDC and profited the gift shop. 

 

Resources and topics covered

 

Emerging and re-emerging infectious diseases:

1. American Society of Microbiology News, Volume 70, Number 6, 2004. "Major resistance mechanisms reported and subsequent development activities."

2. Texas Department of Health protocols for Molecular Diagnostics, including:

            Phenotypic detection of microorganisms using antibody (EIA, Mab, Western     Blot)

            Unamplified genotypic technique-nucleic acid probes

            Amplified signal bDNA nucleic acid probe

            Hybridization probe array        

            DNA amplification; polymerase chain reaction

            Amplification of RNA

3.  TDH - Molecular epidemiology techniques for pathogens

            Archaic and current phenotypic techniques

            Multilocus enzyme electrophoresis (MLEE)

            Genotypic techniques; plasmid analysis, restriction fragment length         polymorphism

            Pulsed-field electrophoreses (PFGE) of genomic DNA with a detailed description         of preparation of bacterial genomic DNA for procedure

            Description of Components of BioRad CHEF Mapper

  1. Suzanne S. Barth, Ph.D., TDH, Austin TX (retired 2003, current e-mail address ssbarth@mail.utexas.edu

      "Not waiting for an outbreak: PFGE for tracking organisms," a detailed comparison of conventional infectious disease epidemiological investigative            techniques with molecular subtyping techniques, including investigations in cases          of Salmonella agona, Listeria monocytogenes, Shigella sonnei.

 

      "Intentional Food Contamination: the Case of the Disgruntled Employee," an      investigation of deliberate food contamination with Shigella dysenteriae 2.

      "Danger at Summer Camp! Non-0157:H7 Shiga toxin producing E. coli (STEC)           Warning! You might miss it!"

      "Waco, Texas, college E. coli 0157:H7 diarrhea outbreak 1994"

      "North TX Investigation 1996"

5.  Biological Weapons: Limiting the Threat, 2000 by Joshua Lederberg, non-fiction            collection on biocrime, bioterrorism.

  • The Demon in the Freezer, 2002 by Richard Preston.
  • "Global Changes Affect Infectious Disease Patterns," ASM News, OL. 62, No. 5, 1996.
  • "Global Climate Change and Emerging Infectious Diseases," Paatz, Epstein, Burke, Balbus. JAMA, January 17, 1996 Volume 275, No. 3.
  • "Cellular Microbiology: How Enteric Pathogens Socialize with Their Intestinal Host," Alessio Fasano, ASM News, Volume 63, Number 5.
  • "Francisella tularensis: an Overview," Titball and Sjostedt, ASM News, Volume 69, Number 11, 2003.
  • "Current Topics: Flu Vaccine Planners Face Challenges on Several Fronts," ASM News, Volume 70, Number 5, 2004.
  • "Wrestling SARS from Uncertainty," Lingappa, McDonald, Simone, Parashar, Emerging Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA. www.cdc.gov/ncidod/EID/vol10no2/03-1032.htm.
  • "Update: Severe Acute Respiratory Syndrome United States, 2003," MMWR, July 4, 2003.
  • "DCD Lab Sequences Genome of New Coronavirus," CDC Office of Communication, Media Relations, www.cdc.gov/od/oc/media/pressrel/r030414.htm.
  • "The risk of transfusion-transmitted prion infections," Rodriguez and Rodriguez, April 2000, www.mlo-online.com.
  • "Antibiotic Discovery: State of the State," ASM News, Volume 70, Number 6, 2004.
  • "Why It Is Important To Continue Antibacterial Drug Discovery," ASM News, Volume 70, Number 6, 2004.
  • "New Strategies for Disigning and Optimizing Vaccines," ASM News, Volume 70, Number 5, 2004.
  • "Clinical Microbiology Eyes Nucleic Acid-Based Technologies," ASM News, Volume 64, Number 2, 1998.
  • "The Application of Molecular Biology to Medical Bacteriology Yesterday, Today, and Tomorrow," ASM News, Volume 65, Number 5, 1999.
  • "The DNA Domain: Molecular Technology Infiltrates the Lab," Deshrain Asthana, November 2000, www.advanceforAL.com.
  • "Studying Microbial Genomes with High-Density Oligonucleotide Arrays," ASM News, Volume 66, Number 8, 2000.
  • "Gen-Probe Amplified Mycobacterium Assays: System Principles," Gen-Probe Incorporated, 9880 Campus Point Drive, San Diego, CA 92121.
  • "Nucleic Acid-Based Tests Move Slowly into Clinical Labs," William Check, ASM News, Volume 67, Number 11, 2001.
  • Manual of Clinical Microbiology, 8th edition, Patrick R. Murray, editor, ASM Press, 2003.

  •       "Molecular Detection and Identification of Microorganisms," Nolte and Caliendo.

  •       "Laboratory Procedures for the Epidemiological Analysis of Microorganisms,"   Soll, Lockhart and Pujol.

  • "Bioterrorism: the ASM Response," ASM News, Volume 68, Number 3, 2002.