I.
MOLECULAR SCIENCE (25-30%)
A.
Nucleic Acid Chemistry
1.
Sugars
2.
Bases
3.
Chemical structure
4.
Associated proteins
5.
Mutations
B.
Basic Molecular Theory
1.
Replication
2.
Transcription
3.
Exons, introns, and splicing
4.
Translation
5.
Chromosome structure
6.
Extrachromosomal structure (e.g.,
phage, plasmid, mitochondrial)
7.
Protein structure
C.
Biochemical Reagents
1.
Polymerase enzymes
a)
DNA
b)
RNA
2.
Endo and exonuclease enzymes
3.
Reverse transcriptase
4.
DNA ligase
5.
Assay development and design
D.
Genetics
1.
Human
2.
Microbial
II.
MOLECULAR TECHNIQUES (20-30%)
A.
Separation and Detection
1.
Electrophoresis
a)
Gel (including agarose acrylamide and
b)
pulsed field)
c)
Capillary
2.
Blotting and probing procedures
(including washing and stringency)
3.
Probe hybridization
4.
Nucleic acid purification
5.
Probe structure (e.g., TaqMan, FRET, simple,
beacon, scorpion)
B.
Nucleic Acid Amplification
1.
Polymerase chain reaction (PCR)
2.
PCR variations (e.g., real-time,
nested, multiplex, arrays, reverse transcriptase)
3.
Branched DNA (bDNA) technology
4.
Sequence based (NASBA)
5.
Transcription-mediated technology (TMA)
6.
Strand displacement amplification (SDA)
7.
Loop-mediated isothermal amplification
(LAMP)
8.
Other (e.g., hybrid capture, ligase
chain reaction, cleavase)
C.
DNA Sequence Analysis
1.
Sanger Sequencing (e.g., chain
terminators)
3.
Automated sequence analyzer
4.
Other (e.g., pyrosequencing)
D.
Other Techniques
1.
Denaturing HPLC
2.
Melt curves analysis
3.
Nucleic acid labeling
4.
in-situ hybridization (ISH)
5.
Restriction fragment length
polymorphism (RFLP)
6.
Epigenetic modification
7.
Array technology (e.g., bead,
microarray)
8.
Multiplex ligation-dependent probe amplification (MLPA)
9.
Mass spectrophotometry
III.
LABORATORYOPERATIONS(15-25%)
A.
Contamination (e.g., biological,
amplified and non-amplified nucleic acid)
1.
Prevention
2.
Monitoring and detection
3.
Elimination
B.
Quality Assurance
1.
A. Specimen collection, preparation, transport and storage
a)
Evaluate quality and quantity of
specimen
b)
Evaluate quality and quantity of
nucleic acid
2.
Reagent selection, preparation
(including calculations), storage, and disposal
3.
Assay selection and validation
4.
Result calculation, interpretation and
reporting
5.
Quality control and proficiency testing
a)
Assay controls
b)
Proficiency testing
6.
Equipment and Instrumentation –
principles, calibration, maintenance, troubleshooting and validation
C.
Guidelines and Regulations
1.
Test System Categories: Analyte
specific reagent (ASR), research use only (RUO), in- vitro diagnostics (IVD)
and lab developed tests (LDT)
2.
Regulations and Standards: CLIA, The
Joint Commission, CAP, CMS, CLSI, FDA
D.
Personnel
1.
Continuing education
2.
Competency
E.
Safety
IV.
APPLICATIONS OF MOLECULAR TESTING
(25-30%)
A.
Infectious Disease
1.
Qualitative Analysis (e.g., MRSA, C. difficile, respiratory pathogens,
STD)
2.
Quantitative Analysis (e.g., viral
load)
3.
Genotypic Characterization (e.g.,
molecular epidemiology, viral typing, resistance testing)
B.
Oncology
1.
Leukemias/Lymphomas (e.g., CML, ALL,
translocations, clonal rearrangements)
2.
Solid Tumors
3.
Hereditary cancer syndromes (e.g.,
breast, colon, ovarian)
C.
Genetics
1.
Hemoglobinopathies (e.g., thalassemias,
sickle cell anemias)
2.
Coagulopathies (e.g., Factor V Leiden,
prothrombin, MTHFR)
3.
Trinucleotide repeat disorders (e.g.,
Fragile X, Huntington, muscular dystrophy)
4.
Single gene disorders (e.g., cystic
fibrosis, Gaucher, hereditary hemochromatosis)
5.
Epigenetic disorders (e.g.,
Prader-Willi, Angelman)
D.
Other
1.
Histocompatibility
2.
Genetic identity (e.g., parentage,
specimen identification, forensic)
3.
Engraftment
4.
Pharmocogenomics (e.g. HerceptinR,
warfarin, PlavixR, carbemazepine)
No comments:
Post a Comment