Module MSE-4089:
Molecular Diagnostics

Overview

The module is delivered over two weeks. One week students will spend 2 and a half-laboratory days preforming an RQ-PCR prognostic test, to determine the level of BCR-ABL transcripts present in peripheral blood-derived from a chronic myeloid leukaemia patient, undergoing standard tyrosine kinase therapy. Two half-day tutorials will then be utilised to develop proficiency in using the BioRad CFX manager qPCR analysis software to perform data quantification and report productions, to the level expected of a certified BCR-ABL monitoring laboratory. European Leukaemia Network

One week students will spend 2 and a half-laboratory performing an immunoprecipitation experiment, to detect 'novel' protein antigenic biomarkers present in peripheral blood-derived from a chronic myeloid leukaemia patient, undergoing standard tyrosine kinase therapy. Two days will then be used to develop proficiency in utilising the BioRad ImageLab analysis software to perform quantitative multiplex western blot analysis to a level required to produce publication-quality data.

Students will employ a number of molecular laboratory techniques; including plasmid extraction, purification, qualitative and quantitative analyse using fluorometer estimation, agarose gel electrophoresis and RQ-PCR analysis of cDNA. Protein work will include standardised extraction of total proteins from human cells, utilising RIPA buffer, estimation of protein concentration using BCA - standard curve analysis, immunoprecipitation utilising magnetic bead technology, SDS-PAGE protein separation and multiplex fluorescent western blotting.

The module is delivered over two weeks. One week students will spend 2 and a half-laboratory days preforming an RQ-PCR prognostic test, to determine the level of BCR-ABL transcripts present in peripheral blood-derived from a chronic myeloid leukaemia patient, undergoing standard tyrosine kinase therapy. Two half-day tutorials will then be utilised to develop proficiency in using the BioRad CFX manager qPCR analysis software to perform data quantification and report productions, to the level expected of a certified BCR-ABL monitoring laboratory.

One week students will spend 2 and a half-laboratory performing an immunoprecipitation experiment, to detect 'novel' protein antigenic biomarkers present in peripheral blood-derived from a chronic myeloid leukaemia patient, undergoing standard tyrosine kinase therapy. Two days will then be used to develop proficiency in utilising the BioRad ImageLab analysis software to perform quantitative multiplex western blot analysis to a level required to produce publication-quality data.

Students will employ a number of molecular laboratory techniques; including plasmid extraction, purification, qualitative and quantitative analyse using fluorometer estimation, agarose gel electrophoresis and RQ-PCR analysis of cDNA. Protein work will include standardised extraction of total proteins from human cells, utilising RIPA buffer, estimation of protein concentration using BCA - standard curve analysis, immunoprecipitation utilising magnetic bead technology, SDS-PAGE protein separation and multiplex fluorescent western blotting.

Assessment Strategy

Threshold (50% / -C) Criterion 1: Knowledge and Comprehension (Subject Specific Expertise) Understanding: The ability to use knowledge in particular, limited contexts, not the capacity to develop or interconnect knowledge Knowledge: The basic foundation of effective learning. Demonstration of knowledge requires and utilizes the powers of memory and recall. Knowledge represents a database of information; facts, principles, ideas and arguments.

Good (-60 / -B) Criterion 2: Analysis, Evaluation & Synthesis (Critical thinking) Analysis: The capacity to dissect information, arguments & ideas. The ability & confidence to stand back & look for logical consistency, completeness, relevance & usefulness. To discover & investigate the basic structure of an idea, reveal hidden meanings, problems & issues. Evaluation: The ability to come to judgments, based upon a critical review of available facts, information & views. The separation of evidence supported facts from unsupported opinions. The use of intellectual problem solving: the willingness, desire & ability to select from competing solutions by systematic evaluation of alternatives. Synthesis: The ability to build on the component parts of an idea or argument in order to develop further ideas. The capacity to engage in constructive, critical assessment of ideas & arguments & appreciate their implication. Originality: To demonstrate & convey ‘self-synthesized’ unique, arguments & discussions.

Excellent (-A / 70%) Criterion 3: Application and Presentation Grade (Transferable skills) The ability to draw upon, and use appropriately, a wide range of knowledge and skills to address questions and issues. The knowledge involved is in part factual, but also includes ideas, concepts, principles, technical expertise or theories Presentation: Written and/or oral and information technology (IT) skills. Indicators include such things as logical structure, coherence and clarity of expression, spelling and grammar, and the use of ICT and numerical data. Where appropriate, also requires that sources are acknowledged and properly referenced and the ability to work effectively as a member of a team.

Learning Outcomes

• Correctly interpret, accurately and ethically report laboratory-derived molecular diagnostic and/or prognostic results

• Critically discuss molecular diagnostic data in the context of medical research

• Demonstrate appropriate good laboratory practices and technical skills for accurate and precise generation of molecular diagnostic data

• Demonstrate in-depth knowledge and understanding of the principles and current limitations of molecular diagnostic applications and data

• Identify and appropriately employ bioinformatic applications to generate, interpret and analyse molecular diagnostic data