Diagnostic Tools Flashcards
(44 cards)
two types of clinical diagnostic tech
molecular and medical imaging diagnosis
technique used to analyze biological markers in the genome and proteome the individual’s genetic code and how their cells express their genes as proteins
molecular diagnosis
technique and process used to create images of the human body for clinical purposes or medical science
medical imaging diagnosis
functions of mass spectrometry
-measures mass better than any other technique and can give info about chem structures
what are mass measurements good for?
to identify, verify and quantitate: metabolites, recomb proteins, proteins isolated from natural sources…etc
how does mass spectrometer work
ion source: makes ions –> mass analyzer: separates ions based on their mass-charge ratio –> mass spectrum: presents info
key specifications of mass analyzers
resolution, mass measurement, accuracy and sensitivity
detection of the various pathogenic mutations in DNA and/or RNA samples in order to facilitate detection, diagnosis, subclassfication, prognosis and monitoring response to therapy
nucleic-acid based molecular diagnosis of human disorders
a genetic disease w/c is caused by a single nucleotide change in the 6th amino acid of the beta chain of hemog.
sicle cell anemia
who and when is the PCR invented
Kary Mullins (1985); noble prize in ‘93
Principles of PCR
- Temperature cycling: denaturation - 94, annealing -55, extension - 72
- every cycle DNA between primers is duplicated
* does exponential amplification
components of PCR rxn
- template DNA
- flanking primers
- thermo-stable polymerase (Taq polymerase)
- dNTP
- PCR buffer
- thermocycler
genetic disorder characterized by abnormal body movements and reduced mental abilities; caused by mutation of Huntingtin (HD) gene
huntington’s disease
individuals w/HD vs. non-HD individuals
non-HD: CAG repeats less than 30x
HD: CAG repeats more than 36x
How can someone determine if they have HD?
PCR can be performed; the DNA is amplified via PCR and sequenced and the number of the trinucleotide repeats is counted
retrovirus that attacks the immune system
HIV
principles of HIV test
HIV tests rely on PCR w. primers that will only amplify a section of the viral DNA found in an infected bodily fluid; pcr product = HIV+, no pcr prod = HIV-
accurate absolute quantification of template molecules by separation of target molecules and counting statistics
droplet digital PCR
Goals of Human Genome Project
- to identify all of the genes in human DNA
- to determine the sequences of the 3 billion bases that make up human DNA
- to create databases
- to develop tools for data analysis
- to address the ethical, legal, and social issues that arise from genome research
implications of genome decipher
epigenetics -> gene regulation -> RNA and protein markers
allows analysis of thousands of gene in one experiment
DNA microarray
Dideoxy (SAnger) method
- denaturation
- primer attachment and extension of bases
- termination
- gel electrophoresis
sequencing that performs large-scale sequencing of DNA by DNA fragmentation, duplication into a vector and individual sequencing
shotgun sequencing (assembled electronically)
NGS Platforms
pyrosequencing sequencing by ligation H+ ion generation/ion torrent reversibly dye terminators massive parallel sequencing illumina flow cell platform
