2/20/2016 OBJECTIVES Popsicle sticks to electrophoresis: how to teach molecular to different levels of students. Audrey Hentzen, PhD, MLS(ASCP)CM Casper College, Casper, WY “One thing seems clear: if all students are helped to the full utilization of their intellectual powers, we will have a better chance of surviving as a democracy in an age of enormous technological and social complexity.” • Describe alternative ways to teach molecular biology • Manipulatives • Dry lab exercises • Online resources • Describe spiral curriculum and its application to teaching clinical molecular techniques at different academic levels. • Compare student learning activities and correlate with ongoing student assessment. GETTING STARTED • Determine academic level of student • Evaluate curriculum • Identify key fundamental concepts that are central themes • Create structure and logical progression of content • Plan and gather resources • Develop assessments to monitor learning success • The Process of Education, Jerome S. Bruner, 1960 • • • • Structure Readiness for learning Intuitive and analytical thinking (spiral curriculum) Motives for learning Gaging student readiness for learning. “match level of these materials to the capacities of the student of different abilities at different grades in school” Bruner, 1960. Level Curriculum Objectives Activities Outcomes Phlebotomy Certificate Program 12 credit hours Entry level: Communication Sociology Computer skills Phlebotomy didactic and clinical practicum Determine and obtain appropriate specimen for collection, processing and knowledge of testing. Patient preparation, specimen collection, and processing. Phlebotomy skills Specimen Collection Processing Patient Care Build upon fundamental curriculum and extend to more complex ideas and content. Level Curriculum Objectives Activities Outcomes MLT AS Degree 60 credit hours General Education Biology Microbiology Math Chemistry Clinical Laboratory Disciplines Specimen collection, analytical testing, QC interpretation, reporting and clinical correlation Laboratory pipetting skills, instrument operation and maintenance, PCR, gel electrophoresis, quality control, data acquisition, manipulation and interpretation. Molecular methodology, patient testing, QC evaluation, data analysis, decision making, and clinical correlation. 1 2/20/2016 Mastery level experiences STRUCTURE OF THE CURRICULUM Level Curriculum Objectives Activities Outcomes MLS BS Degree 120 credit hours Organic Chemistry Biochemistry Cell biology Genetics Evolution Immunology Molecular biology Research Sophisticated analytical testing, skilled techniques, research, patient testing, data management, QC design and development, and clinical diagnostic liaison. Complex, multistep experiments, electrophoresis, blotting techniques, hybridization, detection, data acquisition, management, interpretation, and clinical case studies. Technical skills, direct and amplified testing experience. Use of multiple detection systems and data base utilization for interpretation and management of patient results. Clinical correlation and laboratory resource for physician support. GETTING STARTED • Assess preclinical coursework curriculum • Genetics • Molecular techniques • Microbiology labs • Virology labs • Define fundamentals • Provide enough information to explain the fundamental idea or concept • Provide practice and repetition • Transfer ideas to new situations or problems • Show connections • Allow group interaction for peer tutoring and mastery practice GAUGE YOUR STUDENT LEVELS • Teach at the appropriate level • • • • Phlebotomy MLT MLS DMS • Plan appropriate level learning experiences. • • Assess outcomes Survey clinical environment MLT CONTENT DELIVERY MLT CONTENT DELIVERY • • Internet hybrid delivery Embed molecular content in courses where appropriate. • • • • Microbiology • Content delivery, lectures, assignments and examinations student discussion incorporating clinical laboratory Immunology • Laboratory activities on-campus, for 5 designated lab dates, providing concentrated student laboratory Hematology Immunohematology • Immunology: cellular and molecular mechanisms; host resistance to infectious agents; as well, as hypersensitivities, autoimmunity, tumor and tissue rejection. Includes laboratory for molecular and immunological techniques. • No Designated Molecular Text • Molecular testing taught as a tool used throughout clinical disciplines. practices and procedures will be on-line. activities with one-on-one instruction. • Student laboratory • Molecular manipulatives and genetic databases are used to allow the student to explore the practices of clinical molecular based testing. • Basic PCR amplification, DNA electrophoresis, visualization and documentation. • Each student will complete specified laboratory exercises using basic molecular and immuno- techniques found in clinical laboratories to detect disease states, infectious organisms or patient immune status. • Capstone Activity: • Based upon class discussions, lessons and laboratory experiences, students design an assay system to detect target molecules found in an infectious or disease state. The student describes sample choice, collection, processing, methodology and the overall procedure for the detection of the target molecule from the complex mixture of a human sample. The description includes identification of the target molecule, assay methodology, procedure, detection system, quality control, interpretation of results, and correlation to disease state. 2 2/20/2016 MLT CONTENT DELIVERY- MLT CONTENT DELIVERY- Didactic reinforced with student laboratory. • • Didactic reinforced with student laboratory. Develop fundamental concepts Base Pairing Puzzle Molecular Biology • Review basic molecular cell biology • Identify key concepts that impact molecular testing • • Nucleic Acid Synthesis Protein Synthesis • DNA structure • Base pairing • Unique sequences • • • Collection, handling and storage Target molecules Didactic reinforced with student laboratory. Didactic reinforced with student laboratory. Students evaluate sets of genomic DNA ladders and a standard obtained from suspects or crime scene in case histories provided. Students generate restriction fragment length polymorphism (RFLP) patterns for each individual in the case, analyze and interpret the RFLP patterns to answer case history questions. Southern and northern hybridization DNA fingerprinting Fluorescent in situ hybridization Semi-automated testing Hybridization protection assay Amplified detection system • • • • • • MLT CONTENT DELIVERY- Mathematical expression of PCR Direct nucleic acid testing • • • • • Probe Hybridization Activity Nucleic Acid Hybridization Reactions • Probe design and synthesis • Nucleic Acid Detection • Radioactive labels • Enzyme or Fluorescent labels • Chemiluminescent labels Types of Nucleic Acid MLT CONTENT DELIVERY- Fluorescent Labels • Substrate-labeled fluorescent immunoassay • Fluorescence polarization immunoassay • Microparticle enzyme immunoassay • Radioactive energy attenuation Amplified nucleic acid testing Polymerase chain reaction Ligase chain reaction Nucleic acid based amplification Transcription mediated amplification 5Strand displacement amplification RFLP MAPPING Manipulatives Help to Teach Molecular Diagnostic Concepts, Hentzen, AE (2000), Laboratory Medicine, Sept 2000, Vol 31, Number 9, Pages 513-517. MLT CONTENT DELIVERYDidactic reinforced with student laboratory. • Manipulatives provide free practice of PCR amplification. • Mathematical expression for the PCR amplification. • Given DNA sequences, students identify target sequence, design a reverse primer to specifically amplify a portion of the target DNA sequence. VNTR and STR analysis forward and MLS CONTENT DELIVERY • • cytogenetics, nucleic acid extraction, hybridization, detection, amplification, sequencing, microarrays, and in-situ hybridization. Emphasis is on the areas of the clinical laboratory that use molecular techniques related to genetics, oncology, infectious disease, and identity testing for forensic and transplant purposes. requirements. • Students design reaction conditions for the amplicon hybridization and detection. Students to perform PCR, DNA gel electrophoresis, visualization and documentation. • Case Study PCR Application allows for data interpretation and clinical correlation. • Forensics and paternity testing • Gene mutation detection for disease diagnosis Advanced Clinical Practicum: Molecular • Principles of molecular technology used in clinical laboratories. Laboratory experiences include • Students calculate primer annealing temperature based on the melting • Designated molecular course • Internet hybrid delivery • Content delivery, lectures, assignments and examinations are on-line. There are 5 supplemental laboratory sessions (40 hours) at the UW-Casper Campus and a 24-48 hours of clinical observation at a clinical affiliate. Travel to a clinical affiliate is required for this course. • Lectures and exam will focus on the clinical applications of the various molecular techniques used in diagnostic testing. 3 2/20/2016 MLS CONTENT DELIVERY- MLS CONTENT DELIVERY- Didactic • Didactic • Comprehensive, molecular text: • 2nd Edition (2012), Buckingham, L FA Davis, Philadelphia, PA. • Content: • Fundamentals of Nucleic Acid Biochemistry: DNA, RNA, Proteins • Common Techniques in Molecular Biology: • • • Resolution, Detection, Analysis And Characterization Amplification Inherited Diseases, Oncology And The Classification Of Neoplasms Gene-targeted Therapy Leukemia And Lymphoma Testing Monitoring Response To Therapy Solid Tumor Testing DNA Based Tissue Typing And The MHC Locus • Quality Assurance And Quality Control MLS CONTENT DELIVERYStudent Laboratory Comprehensive, concentrated, on-campus student laboratory sessions. Fundamental experience with application of theory. Common core experience, skills practice and mastery. • Equipment • • • • • • Detection And Identification of Microorganisms • Identity Testing For Forensic And Transplant Purposes Student Laboratory • DNA Polymorphisms • Chromosome Structure And Gene Mutation MLS CONTENT DELIVERY• • • Common Techniques in the Clinical Laboratory • • • • • • • Molecular Diagnostics, Fundamentals, Methods and Clinical Applications. Reagents • • • • • • DNA and Protein Electrophoresis Blotting Visualization Documentation Computer applications Data processing and interpretation • Kits (EDVOTEC) Enzymes Fluorescent labels Gel reagents DNA and Protein molecular marker stains • Unit 1 Module • DNA collection, purification • Restriction digest • DNA electrophoresis • Data bases • DNA Sequence analysis • Sequencing • Translation • Proteonomics • Cloning for sequencing, expression and production of transgenic cells. Laboratory commodities/supplies MLS CONTENT DELIVERY- MLS CONTENT DELIVERY- • • Unit 3 Module Student Laboratory Unit 2 Module • • • • • • • DNA collection, purification • Case Studies Restriction digest • FISH DNA electrophoresis • Cytogenetics Southern blotting Hybridization Visualization Interpretation • Gene expression and in vitro methodologies for expression and detection Student Laboratory • • PCR testing components • Clinical applications Sequence analysis and design of primer (specific and degenerate) • Other amplified techniques • • • • Perform lab • Design and interpretation • Case studies PCR RT-PCR • TMA, NASBA, LCR, PCR-STRS Real time PCR 4 2/20/2016 MLS CONTENT DELIVERY- MLS CONTENT DELIVERY- • • Students are assigned a clinical observation practicum at a clinical affiliate. Student Laboratory Unit 4 Module • • • • • Protein Analysis SDS Page electrophoresis Western Blotting • • • Clinical applications Other Immunoassays Instrumentation • • Immunofixation Interpretation • Design Clinical Correlation Case studies Clinical Observations Students are required to keep a journal of each day’s activities including clinical testing observed, methodology report sheets, and clinical correlation analysis. • Students will rotate through cytogenetics, microbiology, and molecular laboratories. SUMMARY • • • • • • • Simplify first experience and practice to allow for student mastery. Poise attitude for learning confidence and success. Utilize resources for logical progression and building of content complexity. Adjust content for the audience and their scope of practice. Molecular testing is a consideration at all levels of laboratory personnel. Teaching molecular testing as a clinical diagnostic tool in all laboratory disciplines. Utilize alternative methods that reduce programmatic expenses: • Manipulatives • Dry lab exercises • Online resources • Use a spiral curriculum approach, always revisiting fundamentals, and then extend the learning to the next academic and professional level. • • • Allow for the transfer and utilization of concepts in new innovative ways as students progress through curriculum. Focus the student learning activities and correlate with ongoing student outcomes. Assess and evaluate teaching effectiveness for future students. 5