Course

Unit 1

Unit 1:

Microbial Culture and Basic Laboratory Practices, Laboratory safety protocols and biosafety levels, Sample handling, labelling, and documentation, Aseptic techniques and sterile practices Preparation of culture media (selective and differential), Culturing of bacteria: streak plating, pour plates, spread plates, Isolation and identification of microbes using colony morphology and staining (Gram stain), Interpretation of microbial growth patterns

Unit 2

Unit 2:

Nucleic Acid Extraction and Quality Assessment, Principles of DNA and RNA extraction, Manual and kit-based extraction methods from clinical samples (blood, sputum, swabs, etc.), Purification techniques: column-based, magnetic bead-based, Assessment of nucleic acid quality and quantity (Nanodrop, gel electrophoresis), Troubleshooting contamination and degradation, Storage and handling of nucleic acids for downstream applications

Unit 3

Unit 3:

PCR-Based Diagnostics, Principle and setup of conventional PCR, Optimization of reaction conditions (Mg²? concentration, primers, annealing temp), Real-time PCR (qPCR): SYBR Green vs TaqMan assays, Digital PCR: concept, workflow, and clinical relevance, Interpretation of amplification curves and Ct values, Troubleshooting PCR reactions, Documentation, analysis, and reporting of diagnostic results.

Preamble

Diagnostics Lab I is a practical course designed to equip students with hands-on skills in diagnostic laboratory techniques. Students will learn and perform a variety of assays, including microbial culture, DNA/RNA extraction and purification methods, Polymerase chain reaction (PCR) and real-time PCR, digital PCR used in clinical diagnostics. The course emphasizes precision, safety, data recording, and interpretation.

Course outcomes:

CO1: Master microbial culture techniques, aseptic methods, and media preparation.

CO2: Perform DNA/RNA extraction and assess quality using Nanodrop and gel electrophoresis.

CO3: Set up and optimize PCR, qPCR, and digital PCR, and interpret results.

CO4: Isolate microorganisms using morphology, staining, and growth pattern analysis.

CO5: Adhere to safety protocols and maintain accurate lab documentation.

CO6: Troubleshoot and optimize PCR-based diagnostic techniques.

Program outcome

PO1: Bioscience Knowledge

PO2: Problem Analysis

PO3: Design/Development of Solutions

PO4: Conduct Investigations of complex problems

PO5: Modern tools usage

PO6: Bioscientist and Society

PO7: Environment and Sustainability

PO8: Ethics

PO9: Individual & Team work

PO10: Communication

PO11: Project management & Finance

PO12: Lifelong learning

0 – No affinity; 1 – low affinity; 2 – Medium affinity; 3 – High affinity

Program-specific outcome

PSO 1 – Emerging technologies in clinical diagnostics

PSO 2 – Biomolecules in Medicine

PSO 3 – Molecular dysregulation in diseases

PSO 4 – Molecular technology in diagnosis and therapy

PSO 5 – Applying lab discoveries to clinical practice

PSO 6 – Microorganisms in Medicine

PSO 7 – Statistical methods to interpret and validate diagnostic results

PSO 8 – Integrate molecular diagnostics into personalized medicine

PSO 9 – Compounds as biomarkers and its specificity

PSO 10 – Bioinformatics and biological data use

0 – No affinity; 1 – low affinity; 2 – Medium affinity; 3 – High affinity

Textbook:

Mahon CR, Lehman DC, Manuselis G. Textbook of Diagnostic Microbiology. 6th ed. St. Louis (MO): Elsevier; 2018.

Reference Book:

Persing DH, Tenover FC, Hayden RT, Ieven M, Miller MB, Nolte FS, et al. Molecular Microbiology: Diagnostic Principles and Practice. 3rd ed. Washington (DC): ASM Press; 2016.