doc kumareng che Flashcards
science of measuring Ab or Ag in body fluids
serology
production of Ab (substances) that protects the body against the Ag
immune rxn
specialized proteins produced by the immune system in response to the presence of Ag
Ab
molecules (protein or carb) that are recognized by the immune system as foreign of non-self
Ag
enable an early diagnosis or presumptive diagnosis of an infectious dz
Ag test
- identification of a pathogen that has been isolated by culture
- identification of pathogens in different samples
uses of Ab tests
- diagnose a microbial dz when the pathogen microbial Ag is not present in routine specimen
- screen donor blood for different infectious dz
- monitor effectiveness of a given tx by measuring Ab titer
- diagnose autoimmune disorder
immunological techniques that measure the direct interaction b/w Ag and Ab in vitro
primary binding tests (ELISA, IFA, RIA)
immunological techniques that measure the consequences of interaction b/w Ag and Ab in vitro
secondary binding tests (agglutination, CFT, precipitation, neutralization test)
immunological techniques that measure Ag and Ab interactions in vivo (in body)
tertiary binding tests
uses an enzyme system to show the specific combi of an Ag w/ its Ab
ELISA (enzyme-linked immunosorbent assay)
labeled or linked to a specific Ab or Ag
enzyme
added after Ag-AB rxn and acted on (usually hydrolyzed) by the enzyme attached to the Ag-Ab complexes, to give a color change
substrate
4 types of ELISA
- direct
- indirect
- sandwich
- competitive
2 main ways of performing ELISA
- double Ab technique to detect Ag
- indirect technique to detect and assay Ab
PCR applications
- genetic testing
- pathogen detection
- drug development
- crop modification
- forensic analysis
- sequencing
qPCR steps
repeat 25-30 cycles:
1. denaturation (94C)
2. annealing (50-60C)
3. extension (72C)
T/F: qPCR uses a fluorescent dye that only fluoresces when bound to double DNA (dsDNA)
T
T/F: every PCR cycle, more and more ssDNA is made, and fluorescence will get brighter and brighter
F - dsDNAi
T/F: early in the rxn there is so little DNA that the fluorescence cannot be detected
T
it is where fluorescence can observed enough DNA
threshold
cycle at w/c fluorescence is 1st observable above bg
cycle threshold
T/F: a rxn’s Ct is directly linked to the starting concentration of target sequence
T
T/F: in a PCR, the amount of target sequence doubles every cycle
T
T/F: If there is a two-cycle difference, you expect a four-fold difference in starting DNA concentration. A three-cycle difference, eight-fold and so on.
T
