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