Unit 1363: Molecular Diagnostics

1. Introduction to Molecular Diagnostics

• Definition and scope
• Importance in modern healthcare
• Key applications:
  • Detection of genetic variations
  • Infectious disease diagnosis
  • Cancer diagnostics
• Benefits over traditional diagnostic methods

2. Nucleic Acid Amplification Techniques

2.1 Polymerase Chain Reaction (PCR)

• Basic principle and steps (denaturation, annealing, extension)
• Types: conventional PCR, quantitative PCR (qPCR)

2.2 Reverse Transcription PCR (RT-PCR)

• Concept and applications
• Differences from standard PCR

2.3 Isothermal Amplification Techniques

• Loop-Mediated Isothermal Amplification (LAMP)
• Other methods overview (e.g., NASBA, HDA)
• Advantages and limitations compared to PCR

3. DNA Sequencing Technologies

3.1 Sanger Sequencing

• Methodology and chemistry
• Applications and limitations

3.2 Next-Generation Sequencing (NGS)

• Overview of platforms and workflows
• Types of NGS (Illumina, Ion Torrent, etc.)
• Applications:
  • Mutation identification
  • Gene expression analysis
  • Microbial genome characterization

4. Molecular Biomarkers

• Definition and classification
• Types of biomarkers:
  • Genetic markers (e.g., SNPs, mutations)
  • Protein markers
  • microRNA signatures
• Role in:
  • Disease diagnosis
  • Prognosis
  • Treatment monitoring
• Examples of clinically relevant biomarkers

5. Infectious Disease Diagnostics

• Molecular detection of pathogens:
  • Bacteria
  • Viruses
  • Parasites
• Techniques:
  • PCR and real-time PCR assays
  • Nucleic acid probes
  • Sequencing for pathogen identification
• Advantages of molecular methods in rapid and accurate diagnosis

6. Cancer Molecular Diagnostics

• Molecular markers in cancer biology:
  • Oncogenes and tumor suppressor genes
  • Mutation analysis
• Liquid biopsies and circulating tumor DNA (ctDNA)
• Personalized medicine approaches
• Monitoring treatment response and disease progression

7. Quality Control and Assurance in Molecular Diagnostics

• Importance of quality control (QC) and quality assurance (QA)
• Validation procedures for assays
• Regulatory requirements and guidelines
• Proficiency testing and external quality assessment (EQA) schemes
• Best laboratory practices

8. Emerging Trends in Molecular Diagnostics

• Point-of-care testing (POCT)
• Digital PCR technology
• CRISPR-based diagnostic tools
• Integration of artificial intelligence (AI) and machine learning for data analysis
• Future perspectives and innovations