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Virology > Week 2 > Flashcards

Week 2 Flashcards

1
Q

Can viruses multiply by division?

A

No, they do not have the genetic capability to do so.

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2
Q

What do viruses need to replicate?

A

A living host

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3
Q

What dose a virus do once inside the host cell?

A

The virus hijacks & utilizes the host cell machinery to produce its proteins & nucleic acid for the next generation of virus

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4
Q

What are 3 ways we can grow/cultivate viruses outside of the living host?

A
  • Cell culture/tissue culture - Inoculation in embryonated egg - Use laboratory animals
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5
Q

What is the best way to grow viruses (even though it isn’t ethical)?

A

Actual host itself

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6
Q

Most sophisticated type of cell/tissue culture…

A

Organ culture

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7
Q

Organ Culture:

A

Section of organ cultured in-vitro. New method & technical difficulties. Not very popular method.

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8
Q

One of the best ways of cultivating viruses…

A

Cell Culture

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9
Q

Cell Culture:

A

Removal of cells from living host where cells are then grown in an artificial environment– where we inoculate the virus into the growing cells — where the virus will infect the host cells in-vitro & increase in number

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10
Q

What are the 2 types of cell culture?

A

1) Suspension Cultures 2) Monolayer Cultures

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11
Q

Suspension Cultures:

A

Where cells are suspended/floating in a liquid medium

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12
Q

Monolayer Cultures:

A

Most common, cells are grown on plastic surface where they grow in a 1 layer, where viruses are grown. CELLS ARE ATTATCHED

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13
Q

We take some of my liver cells. Cut it in small pieces, then use enzymes to break into individual cells, then into growth media, then cells start growing in a Monolayer fashion. Once there is one layer– known as contact inhibition.

A

Primary Cell Culture

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14
Q

Heterogeneous culture

A

Primary Cell Culture

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15
Q

What culture is used for producing viral vaccines?

A

Primary Cell Culture

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16
Q

What are the disadvantages of Primary Cell Culture?

A

-Difficult to obtain -Short lifespan in culture -Very susceptible to contamination -May not fully act like parent tissue due to complexity of culture media

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17
Q

Secondary Cell Culture:

A

When a primary cell culture is sub-cultured.

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18
Q

Subculture (or passage):

A

Transfer of cells from one culture vessel to another culture vessel

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19
Q

For further propagation of cells, we need more space so we do a _________ _________.

A

Secondary cell culture / Sub-culture

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20
Q

What are the 2 types of Cell Lines?

A

1) Finite / Diploid Cell Lines 2) Continuous Cell Line

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21
Q

_______ Cell Lines means cells can only be propagated up to a certain extend. Their lifespan is limited.

A

Finite/Diploid Cell Lines

22
Q

__________ Cell Lines means they have the ability to divide indefinitely. Edward Cullen of cell lines.

A

Continuous Cell Lines

23
Q

Example of HeLa cells:

A

Continuous Cell Line / Cancer cells

24
Q

Growth Medium %:

A

5-10%

25
Q

Maintenance Medium %:

A

0-2%

26
Q

Phenol Red is used as a…

A

pH indicator

27
Q

-Contamination -Metabolic waste that accumulates from the healthy cell

A

Phenol Red is used to sense a pH changes

28
Q

Most mammalian cells like a pH of…

A

7.0 - 7.4

29
Q

What do most virologist add to cell culture media before propagating growth?

A

Antibiotics/Antimicrobials

30
Q

Human & Mammal cell temp:

A

36c-37c

31
Q

Insect cell temp:

A

27c

32
Q

Avian cell temp:

A

38.5c

33
Q

If cells are left in Trypsin for too long…

A

Cells become damaged and die

34
Q
A

Positive stained cells in Adenovirus shell vial culture

35
Q

How are viral antigens detected using the shell vial technique?

A

Using a pool of fluorescein-labeled monoclonal antibodies

36
Q
A

No visible development / Infertile egg

37
Q
A

Developing embryo in the center of the developing vascular system / Fertile egg

38
Q
A

Dead embryo / blood ring indicates early embryonic death

39
Q

What do ultracentrifuges provide to virology?

A

Sufficient gravitational force 60,000 RPM and 200,000 x g to 150,000rpm and 1,000,000 x g to sediment the smallest of viruses.

40
Q

Rate-Zonal Centrifugation:

A

Sucrose gradient centrifugation

41
Q

Isopycnic Centrifugation:

A
  • Buoyant Density
  • Isopycnic Point
42
Q

What is the most effective Assay?

A

Monolayer Plaque Assay

43
Q

What is a pock of a pock assay?

A

Pock= necrotic area on chorioallantoic membrane (CAM) of embryonated egg

44
Q

What is a transformation assay?

A

It is used for oncogenic viruses, transformed cells lose contact inhibition and create “piles” of cells, focus forming units

45
Q

What is a quantal assay?

A

It measures the precence of absence of a virus/infection. Used for viruses that don’t form plaques. Endpoint is virus dilution that affects 50% of the test subjects

46
Q

___________ is the tissue culture infectious dose which will infect 50% of the cell monolayers challenged with the defined inoculum

A

TCID / Tissue Culture Infectious Dose

47
Q

________ is the average number of virus particles infectiong each cell.

A

MOI / Multiplicity of Infection

48
Q

Calculation of TCID50 using the Reed & Muench Method

A

% infectious about 50% - 50%

DIVIDED BY

% infectious about 50%. - % infection below 50%

49
Q

What is a ultracentrifuge used for?

A

To purify and concentrate viruses

50
Q

What are the 2 types of ultracentrifugation?

A
  • Rate-Zonal
  • Isopycnic
51
Q

Which centrifugation type uses a sucrose gradient?

A

Rate-Zonal

52
Q

________ centrifugation has a uniform mixture of smaple and a density gradient such as Caesium Chloride.

A

Isopynic

Virology (5 decks)

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