(07) Vaccines Flashcards
these are the main points he wants us to get out of these lectures
just look at this
(Electron Microscopy)
- negative staining (contrast stain)
- immuno electron microscopy
1-2. What are the two limitations?
- large number of particles required (sensitivy is bad)
- viruses with similar morphology difficult to distinguish
(advantage - can see what virus looks like by looking at its morphology - get general idea)
(Virus Isolation)
- able to detect, and get a isolate of the virus out of the sample
1. What is the advantage of being able to isolate?
2. disadvantage?
3. Which viruses don’t work
4. which do?
- can further characterize it
- not all viruses can be isolated (do this in embryonic eggs and cell cultures - if virus can’t grow on these you are screwed)
- Hantavirus and retrovirus
- herpes
(Virus isolation from cell culture)
Two types
- prepared directly from tissue from an animal or embryo
- transform immortalized cell lines, can be passaged indefinitely
- What are changes in the cells in the culture referred to as? what do some viruses produce?
- What is an anooying this about this?
- primary cell culture
- continuous cell culture
- cytopathic effect (CPE); inclusion bodies
- takes a long time
(Test for confirming Virus Specificy in Virus Isolation)
- immunoflouoresnence
- immunohistochemistry
- PCR
- hemadsorption
- hemagglutination
(Localization of Viral Antigen in Tissues)
1-2. What two methods allow you to test for virus antigen in particular tissues?
- Immunofluorescence
- Immunohistochemistry
(Immunofluorescence Assays)
- Ab is labeled with what?
- What is the most common dye?
- When FITC binds to antibodies does it affect their reactivty?
- what color does it show as?
- fluroscent dye
- FITC
- no
- green
(Direct vs indirect)
What is the
C
(ELISA)
Enzyme linked immunosorbent assay
- used for antibody or for antigen?
- can be either
Hemaggluination ASSAY
only works for a few different types of viruses
(Detection of Viral Nucleic Acids)
- What is this called (the most common one used for the detection of a particular virus)?
- work on RNA or DNA?
- What’s the limitation here?
- qualitative or quantitative?
- PCR
- can do both
- have to be looking for specific virus
- can be both
(Virus Quantitation)
- qualitative = yes/no
- quantitative = how much
examples of quant
(titration - serial ten fold dilutinos of virus)
(plaque reduction assay)
(quantitative PCR - Taqman)
D
(All of these are tests to detect the virus)
(PCR is the most sensitive)
B
- Why would you ever want to test for antibody?
- Should see a rise in what following virus exposure?
- What does DIVA differentiate?
- What does wurveillance accomplish?
- seeing if a virus has been exposed to something
- antibody titer
- infected from vaccinated
- detects prevalence (looking at a population)
in ELISA for FeLV and FIV
looking for FeLV antigen and FIV antibody
based on gel immunodiffusion
R - reference positive samples
A - neg
B - pos
C - neg
wanted (green) and unwanted (red) reactions
