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Virology > Class-Cult-Purif-Quant (Ex1) > Flashcards

Class-Cult-Purif-Quant (Ex1) Flashcards

1
Q

What are the classes in the Baltimore Classification System?

A 
Class1: DNA double stranded
Class2: DNA single stranded (+)
Class3: RNA double stranded
Class4: RNA single stranded (+)
Class5: RNA single stranded (-)
Class6: DNA int. RNA single stranded (+)
Class7: RNA int. DNA double stranded
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2
Q

What does the ICTV system consider for classification of viruses?

A
  • nature of virus genome and virus genetic diversity
  • virus replication strategies
  • virus morphology
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3
Q

What does ICTV stand for?

A

International Committee on Taxonomy of Viruses

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4
Q

What are the suffixes for the ICTV System?

A

Order - virales
Family - viridae
Subfamily - virinae
Genus - virus

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5
Q

What are suspension cultures?

A
  • cells which do not require attachment for growth or do not attach to the surface of the culture vessels
  • can be propagated in suspension
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6
Q

What are monolayer cultures?

A

when the bottom of the culture vessel is covered with a continuous layer of cells, usually one cell in thickness

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7
Q

What is a primary cell culture?

A
  • cells dissociated directly from the parental tissue of origin
  • cells have same number of chromosomes as parent tissues
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8
Q

Disadvantages of primary cell culture

A
  • difficult to obtain
  • short lifespan in culture
  • susceptible to contamination
  • may not act like parent tissue
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9
Q

What is secondary culture?

What is a cell line?

A
  • transfer culture
  • primary culture is sub-cutlured
  • transfer of cells from one culture to another
  • after first subculture, primary culture becomes a cell line
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10
Q

Describe a fibroblastic cell

A
  • bipolar or multipolar
  • elongated shape
  • grow attached to a substrate
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11
Q

Describe an epithelial-like cell

A
  • polygonal shape

- grow attached to a substrate in discrete patches

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12
Q

Describe a lymphoblast-like cell

A
  • spherical in shape

- grown in suspension without attaching to surface

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13
Q

Describe the Shell Vial technique

A
  • cover slip with monolayer of cells
  • put in media in vial, add virus, centrifuge
  • when virus has grown, take out cover slip and add antibodies with fluorescent dye
  • antibodies bind to virus
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14
Q

What are the routes of inoculation of an egg?

A
  • Yolk sac inoculation
  • Allantoic cavity inoculation
  • Amniotic cavity inoculation
  • Chorioallantoic membrane inoculation
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15
Q

Explain Rate-Zonal Centrifugation

A
  • sample is layered
  • under centrifuge, heavier particles move to the bottom
  • particles of each size settle as discrete bands
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16
Q

Explain Isopycnic Centrifugation

A
  • particles separated on the basis of their buoyant density

- isopycnic point is when the buoyant density of a particle equals that of the medium

17
Q

What is the virus titer?

A

the lowest concentration of virus that still infects cells

18
Q

Virus Counter 2100

A
  • measures intact virions through detection of protein and nucleic acids
  • each sample stain with two different dyes, one for nucleic acid and one for protein
  • analyzed as they flow through a laser beam
19
Q

Hemagglutination Assay

A
  • add virus to RBCs in suspension

- positive: RBCs become linked by ND virus receptors and form a mat

20
Q

What is a plaque?

A

a circular zone of necrotic cells surrounded by viable cells in a monolayer

21
Q

Monolayer Plaque Assay

Units?

A
  • measures the number of virus particles capable of forming plaques per unit volume
  • plaques will be colorless
  • plaque forming units/mL
22
Q

Calculation for determination of the Titer(PFU/mL) for Monolayer Plaque Assay

A

Average titer divided by dilution

23
Q

Pock Assay

Units?

A
  • necrotic area on chorioallantoic membrane of embryonated egg
  • pock-forming units/mL
24
Q

Transformation Assay

Units?

A
  • determination of titers or oncogenic viruses
  • transformed cells lose contact inhibition and heap upon each other
  • focus-forming units
25
Q

Quantal Assay

A
  • measures presence or absence of infection

- endpoint: virus dilution that affects 50% of test subjects

26
Q

Describe Finite/Diploid Cell lines

A
  • limited life spans and go through a limited number of cell generations
  • derived from embryos or secondary cultures
  • cells retain original morphology and diploid
  • growth rate slow
  • contact inhibition, anchorage dependence
27
Q

Describe Continuous Cells lines

A
  • divide indefinitely
  • derived from cancer cells or induced transformation of primary strain
  • genetically furthest from animal
  • abnormal morphology and chromosome number
  • rabid growth rate
  • lack of contact inhibition and anchorage dependence

Virology (15 decks)

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