Molecular Methods in Clinical Microbiology (A Historical Review)

Continuing Education Credits

• P.A.C.E.® Contact Hours: 1.5 hour(s)
• Florida Board of Clinical Laboratory Science CE Credit Hours -
  General (Microbiology/Mycology/Parasitology): 1.5 hour(s)

Objectives

• Describe some of the history of molecular methods and their introduction into the routine diagnostic laboratory.
• Describe some of the advantages of molecular methods over traditional microbiology.
• Describe the requirements and some of the challenges of implementing molecular methods in the setting of a routine clinical microbiology laboratory.
• Describe the principles of the basic methods of molecular techniques.
• Describe several assays of interest (and principles of reaction) for use in the field of infectious disease.

Course Outline

• Some History of Development
• Prior to 1985
• Chlamydia trachomatis and Neisseria gonorrhoeae
• Human Papilloma Virus (HPV) and Mycobacterium
• Hepatitis and Viral Load Testing
• Initially, why were molecular methods difficult to perform in routine clinical laboratories?
• Why were Chlamydia and Neisseria logical targets for the development of a commercial molecular assay?
• Potential Benefits of Molecular Methods over Traditional Microbiology
• The Key Benefits: Improved Sensitivity of Detection
• The Key Benefits: Improved Sensitivity of Detection, continued
• The Key Benefits: Specificity of Identification
• The Key Benefits: Reduced Turnaround Time
• In traditional culture or antigen detection methods, the sensitivity of detection is adversely affected by which of the following?
• Why can molecular methods offer improved turnaround times over-cultivation methods?
• Challenges for Implementing Molecular Microbiology
• Challenges for Implementation: Space Requirements
• Challenges for Implementation: Separation of Key Activities
• Challenges for Implementation: Workflow Requirements
• Challenges for Implementation: Required Work Skills
• Challenges for Implementation: Cost
• Why is it important to consider work space and workflow design for molecular methods?
• Molecular testing entails precise workflow requirements. Technologists must progress through a series of steps in a specific order to ensure quality r...
• Definitions and Principles of Basic Methods
• Methods: An Overview
• Categories of Methods
• Polymerase Chain Reaction (PCR)
• Reverse Transcriptase Polymerase Chain Reaction (RT-PCR)
• Detection and Identification of Polymerase Chain Reaction (PCR) Products
• Detection and Identification of Polymerase Chain Reaction (PCR) Products: Advantages of Real-Time PCR
• What are the two general categories of nucleic acid amplification test (NAAT) techniques?
• Polymerase chain reaction (PCR) copies DNA through repeated cycles of three basic steps. What is the correct order of these steps? (Choose the BEST an...
• Application to Infectious Diseases
• Assays of Interest
• Assays of Interest for Infectious Disease [Staphylococcus aureus and Methicillin-resistant S. aureus (MRSA)]
• Identification of Staphylococcus aureus with Peptide Nucleic Acid (PNA)-Fluorescence In Situ Hybridization (FISH)
• Detection and Identification of Methicillin-resistant Staphylococcus aureus (MRSA) by Polymerase Chain Reaction (PCR)
• Development of Assays
• Molecular Versus Culture - Pros and Cons
• Current and Future Prospects
• What is successful molecular identification of methicillin-resistant Staphylococcus aureus (MRSA) based upon? (Choose the BEST answer)
• Potential disadvantages of molecular methods for methicillin-resistant Staphylococcus aureus (MRSA) include:
• Assays of Interest for Infectious Disease [Influenza and Other Respiratory Viruses]
• Influenza: Prior Traditional Methods and the Need for Change
• Introduction of Molecular Methods
• 2009 H1N1 Influenza (Swine Flu)
• Incremental Improvements for Influenza Testing
• 2019 SARS-CoV-2 (COVID-19)
• Which statement about the 2009 H1N1 virus is TRUE?
• Which statement is TRUE about the molecular methods made available under the Emergency Use Authorization (EUA) during 2009 H1N1?
• Assays of Interest for Infectious Disease [Clostridioides difficile]
• Clinical Significance of Clostridioides difficile
• Previous Methodologies: Culture and Cell Cytotoxicity Neutralization Assay (CCNA)
• Other Methodologies: Antigenic Detection of Toxin and Glutamate Dehydrogenase (GDH)
• Molecular Methods
• Example #1
• Example #2
• Other examples...
• Several methods of detection are available for the detection of Clostridioides difficile (previously known as Clostridium difficile) in clinical sampl...
• What statement is TRUE about the glutamate dehydrogenase (GDH) assay for Clostridioides difficile?
• The Future of Development
• The Future ...
• References
• References

Additional Information