OVERVIEW

Molecular Methods in Clinical Microbiology is organized by MediaLab, Inc..

Course description:
This course offers a historical look at the progression of molecular methods used in the clinical laboratory. The advantages of these molecular methods over traditional microbiology are discussed, along with the requirements and challenges faced during implementation in a routine clinical setting. Basic methods and molecular techniques are described, including the principle reactions of some assays of current interest for infectious diseases.

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