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Liz's Serology/Virology Module 1 > Immunofluorescence and Viral Disease Diagnosis > Flashcards

Immunofluorescence and Viral Disease Diagnosis Flashcards

1
Q

What is a method for efficient and definitive detection of viral Ags in virus-infected cells?

A

Immunofluorescence (FA) staining in virology

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2
Q

3 applications of FA

A
  • Transport medium sediment from samples collected on swabs
  • Cells from infected cell culture monolayers
  • Shell vial monolayers
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3
Q

Procedure of immunofluorescence staining

A
  1. Cells are fixed to a slide
  2. Monoclonal Abs applied in direct or indirect staining protocols
  3. One of the Ab preparation is labeled w/ a fluorescence dye
  4. Results are red w/ a fluorescence microscope
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4
Q

FA stain: fluorescein isothiocyanate (FITC) is used most

  • What color does it fluoresce?
  • What does it stain?
A
  • Fluoresces bright green

- Stains the target

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5
Q

FA stain: Evans blue is used as a counterstain

  • What color does it fluoresce?
  • What does it stain?
A
  • Fluoresces red

- Stains the background

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6
Q

Steps in direct FA staining?

A
  1. Fluorescein-labeled Abs are added to a microscope slide w/ smear of virus-infected cells
  2. Incubate and rinse
  3. If Ag/Ab binding, then green fluorescence
  4. If no binding, no fluorescence
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7
Q

Steps in indirect FA staining?

A
  1. Abs added to microscopic slide w/ smear of virus-infected cells
  2. Incubate and rinse
  3. Stage 1: there will either by Ag/Ab binding or not
  4. Stage 2: fluorescein-labeled anti-species globulin added
  5. Incubate and rise
  6. If Ag/Ab binding, then fluorescence
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8
Q

In FA testing, what are smears evaluated for?

A

Intensity and distribution of fluorescence

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9
Q

How do we stain cells from a cell culture monolayer?

A
  • Use pipette/scraper to scrape cells off tube wall
  • Spin to sediment the cells
  • Make smear of sediment
  • Stain, read
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10
Q

How do we stain cells from cells collected on swabs submitted in transport medium?

A
  • Use sediment found in transport medium after centrifugation
  • Make a smear on a microscope slide
  • Stain and read (at least 20-25 cells must be present to be valid test)
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11
Q

Enzyme-linked virus inducible system (ELVIS) is specific for what virus?

A

Herpes simplex virus (HSV)

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12
Q

How does ELVIS work?

A
  • E. coli LacZ gene is cloned into cells behind an HSV promoter
  • A substrate is added to infected cells. If beta galactosidase enzyme is present (only produced by HSV infected cells), it acts on the substrate to produce a color change (DARKER BLUE)
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13
Q

Advantages to ELVIS

A
  • Very little skill required
  • Expensive monoclonal Abs not needed
  • Sensitive and specific compared to cell culture
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14
Q

Disadvantage to ELVIS

A

Virus-specific

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15
Q

2 ways antivirals work

A
  • Nucleoside analogs (allows virus to add it in and then destroy it)
  • Enzyme inhibitors
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16
Q

Are antivirals “virus specific” or “broad spectrum”?

A

Virus-specific

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17
Q

What is CMV’s antiviral?

A

Gangciclovir

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18
Q

What is HSV’s antiviral?

A

Acyclovir

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19
Q

What is HIV-1’s antiviral?

A

Didanosine

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20
Q

What is RSV’s antiviral?

A

Ribavirin

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21
Q

What test involves using components labeled w/ active enzymes?

A

Enzyme immunoassay EIA Ag detection

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22
Q

What is EIA used for?

A

To detect unknown Ags or Abs (widely used for testing fecal samples for rotavirus Ag)

23
Q

Strains of influenza A vary in regards to what?

A

Hemagglutinin (H) and neuraminidiase (N)

24
Q

Herpesviridae family includes…

A
  • Herpes simplex virus, types 1 and 2
  • Cytomegalovirus (CMV)
  • Varicella-Zoster virus
  • Epstein Barr virus
25
Q

Herpesviridae family includes ____ viruses associated w/ various syndromes. All establish ____

A

DNA; latency

26
Q

Usually causes oral sores

A

HSV type 1

27
Q

Usually causes genital sores

A

HSV type 2

28
Q

Most common STD

A

HSV types 1 and 2

29
Q

Produces vesicular lesions, stress reoccurrence is common, congenital infections are the most severe manifestation

A

HSV, types 1 and 2

30
Q

HSV, types 1 and 2

- Virus isolation

A

Grows in 24-48 hours in several cell lines

31
Q

HSV, types 1 and 2

- Ag detection

A

Sensitivity good in moist lesions, otherwise not very sensitive (as low as 50%) compared to cell culture

32
Q

HSV, types 1 and 2

- Serodiagnosis

A
  • Not useful in diagnosis, cross reactivity b/w HSV-1 and HSV-2
  • Used to determine HSV “immune status”
33
Q

HSV, types 1 and 2

- Molecular methods

A
  • Excellent

- Method of choice, especially for CSF

34
Q

HSV, types 1 and 2

- Treatment

A
  • Acyclovir, Foscarnet, others

- Very effective, few side effects

35
Q
  • Asymptomatic/mild infections in most healthy (immunocompetent) individuals
  • Can cause a mononucleosis-like syndrome (resembles classic infectious mono clinically) in healthy people
  • Recurrence during stress is common
  • Congenital infections are possible, especially w/ new infections during pregnancy
  • Severe infections in immunocompromised individuals
A

Cytomegalovirus (CMV)

36
Q

Cytomegalovirus (CMV)
- Virus isolation
-

A
  • 3 weeks or longer in traditional cell cultures

- faster results in shell vials, sensitivity varies w/ type of specimen

37
Q

Cytomegalovirus (CMV)

- Ag detection

A
  • Generally NOT useful
38
Q

Cytomegalovirus (CMV)

- Serodiagnosis

A

Useful in diagnoses (seroconversion or IgM-specific detection) and for immune status testing

39
Q

Cytomegalovirus (CMV)

- Molecular diagnositcs

A

Excellent, especially for quantitative determinations

40
Q

Cytomegalovirus (CMV)

- Treatment

A

Ganciclovir

41
Q
  • Agent of chickenpox, shingles
A

Varicella-Zoster virus (ZVZ)

42
Q

Varicella-Zoster virus (ZVZ)

- Virus isolation

A

6-8 days in standard cell cultures

43
Q

Varicella-Zoster virus (ZVZ)

- Ag detection

A

Highly sensitive in smears from moist lesions

44
Q

Varicella-Zoster virus (ZVZ)

- Serodiagnosis

A

Not useful in diagnosis, “immune status” only

45
Q

Varicella-Zoster virus (ZVZ)

- Molecular methods

A
  • Very sensitive and accurate

- Not widely available b/c other less expensive methods are used → Ag detection

46
Q

Varicella-Zoster virus (ZVZ)

- Treatment

A
  • Sometimes not treated

- High dose of acyclovir for serious cases

47
Q

Agent of infectious mononucleosis

A

Epstein Barr virus

48
Q

Epstein Barr Virus

- Ag detection

A

Not available

49
Q

Epstein Barr virus

- Virus isolation

A

Does not proliferate in standard cell cultures

50
Q

Epstein Barr Virus

- Molecular diagnosis

A

Other methods are used → Ag detection

51
Q

8 respiratory viruses detected by FA in respiratory viral Ag profile

A
  • Adenovirus
  • Human metapneumovirus
  • Influenza A and B
  • Parainfluenza 1-3
  • Respiratory Syncytial Virus (RSV)
52
Q

Appearance of positive enzyme-linked virus inducible system

A

Pos: Beta galactosidase enzyme is present and acts on the substrate causing a color change

53
Q

Strains of influenza A

A

H5N1, H7N9, H1N1, vH3N2

Liz's Serology/Virology Module 1 (5 decks)

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