Chapter 2 Flashcards
Propogation
Growth
need a specific host
Animal Cell Culture
Continuous Cell Line
Immortal Cells
Plaque
A clear area in host lawn
Contains Genetically Identical viral particles
Differential Centrifugation
2-step centrifugation,
Low speed
High Speed
Density Gradient Centrifugation
for further purification
Use a series of solutions at different concentrations
Types of Density Gradient Centrifugation
Rate Zonal centrifugation
Equilibrium centrifugation
Rate Zonal Centrifugation
lay viral prep on top of pre-formed gradient
Equilibrium Centrifugation
Suspend virus in a solution of gradient materials
Electron Microscopy
High magnification and resolution
Negative staining with heavy metal
High cost and low sensitivity
Infectivity
Capacity of a virus to replicate in host cell
CPE
Cytopathic Effect
Visible change in host cell due to viral infection
Syncytium
cells fuse-> multinucleated cell
Detect Viral Antigens
Direct and indirect method
Hybridization
Detecting viral nucleic acid
Use labeled sequence-specific DNA probe
Hybridization techniques
Southern blotting for DNA
Northern Blotting for RNA
in situ directly use thin section of tissue
PCR
Polymerase Chain Reaction
Use viral specific primers to amplify viral DNA
for low number of copies
RT-PCR
Revers Transcriptase PCR
SDS-PAGE
Sodium Dodecyl Sulfate Polyacrylamide Gel Elecrophoresis
Analysis of viral proteins
Infectivity Assays
measure the titer of infective virus in a sample
Plaque Assay
A quantitative assay to determine viral titer
plaque forming unit
also used to isolate and purify viruses
Quantal Assay
Determine viral dose ID50 LD50 TCID50 Look for CPE or other change
One-Step Growth Curve
Adsorption
Latent(eclipse) period
Rise Period(burst)
Burst size
Average Number of virions released from an infected cell
final viral titer/initial viral titer
MOI
Multiplicity of Infection
# of virions/# of cells
helps to determine viral load
