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MICRO2209 > virus detection and cultivation > Flashcards

virus detection and cultivation Flashcards

1
Q

What are specimens used to culture or detect viruses?

A 
throat swabs
nasopharyngeal aspirates
vesicle fluid
blood
cerebrospinal fluid
saliva
biopsy
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2
Q

What is important during transport of samples?

A

temperature: 4 degrees, -20 degrees, -80 degrees, -150 degrees

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3
Q

What are the two

A
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4
Q

What are some examples of direct detection?

A

electron microscopy
isolation of infectious virus by cell culture
detection of viral genome by molecular methods
detection of virions by serological methods

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5
Q

What is indirect detection?

A

measurement of virus-specific host immune response after infection

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6
Q

Why is electron microscopy used instead of light microscopy?

A

can be readily identified

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7
Q

How can viruses be isolated?

A

by propagation in animals or human cells

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8
Q

What is the most common method for virus propagation?

A

cell culture

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9
Q

How does cell culture work?

A

tissues dissociated into single cells by mechanical disruption and proteolytic enzymes
cells suspended in culture medium and placed in culture vessels

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10
Q

What is cell culture medium made of?

A

isotonic solutions of salts, glucose, vitamins and amino acid @ 7.2-7.4 pH

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11
Q

What are the 3 main kinds of cell cultures?

A
  1. Primary cells (5-10 cell divisions)
  2. Diploid cells (>100 divisions)
  3. Continuous cell lines (infinite)
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12
Q

What are evidence of viral growth?

A

cytopathic effects visible - inclusion bodies / morphological alterations

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13
Q

How can we detect viral genomes?

A

via PCR

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14
Q

How is viral antigen detected?

A

using immunofluorescence

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15
Q

How does indirect IFA work?

A

the antibody is labelled with a flurophore

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16
Q

How does indirect IFA work?

A

a second antibody which recognised the Fc region of the first antibody, is labelled with a flurophore

17
Q

What is enzyme-linked immunosorbent assay?

A

a way to detect viral antigen or antiviral antibody

18
Q

How does enzyme-linked immunosorbent assay work?

A
  1. antigen fixed onto a solid phase
  2. antigen or antibody of interest is allowed to bind
  3. detector antibody (conjugate) is added
  4. enzyme substrate added
  5. enzyme/substrate combinations produce a colour change
19
Q

What is a way to measure infectious virus?

A

plaque assay

20
Q

What are plaque assays?

A

area of killed cells in a tissue cultured vessel; visible damage can be tested
each infectious virus particle produces a circular zone called plaque

21
Q

What is haemagglutination?

A

identifying the grouping of RBC caused by viral antigen (attachment to sialic acid)

22
Q

What is a Western blot?

A

separation based on size, then transferred to nitrocellulose membranes (blotting) and viewed

MICRO2209 (16 decks)

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