Testing of Viruses Flashcards

1
Q

Conventional cell culture cell monolayer

A

Growing cell monolayer (single confluent layer) on plastic or glass tube

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2
Q

Conventional cell culture incubation conditions

A

Roller drum, tilted 5-7 degrees, revolving at 0.5-1 rpm at 35-37°C for 1 to 4 weeks

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3
Q

Advantage of conventional cell culture

A

Can detect different viruses in one cell culture

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4
Q

Disadvantage of conventional cell culture

A

Long, laborious process

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5
Q

EDTA/Heparin blood handling

A

Separate buffy coat

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6
Q

CSF specimen handling

A

Inoculate 1 mL directly

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7
Q

Feces/Rectal swab handling

A

Place in 2 mL VTM → Vortex → Centrifuge → Inoculate

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8
Q

NPS/OPS/Sputum/Respiratory washings handling

A

Dilute with VTM

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9
Q

Tissue specimen handling

A

Mince with scalpel/scissors → Gently grind → Suspend in VTM → Centrifuge → Inoculate

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10
Q

Swab/Fluid specimen handling

A

Emulsify in VTM

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11
Q

Urine specimen handling (clear)

A

Inoculate directly

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12
Q

Urine specimen handling (turbid)

A

Centrifuge → Inoculate

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13
Q

Genital/Skin (Vesicle fluid/Scrapings) specimen handling

A

Emulsify in VTM

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14
Q

Urine cell culture for adenovirus

A

HDF, HEp-2

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15
Q

Major cell culture types

A

PMK, HDF, HEp-2

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16
Q

Genital/Skin cell culture

A

HDF

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17
Q

Cell culture tube procedure

A

Absorption (12-24 hours) → Maintenance tube (replace 1-2x/week) → Incubation (5-28 days)

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18
Q

Freezing in cell culture

A

Remove old medium → Add fresh culture medium

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19
Q

Passaging or Splitting

A

Remove cells from surface, dilute, place into new container

20
Q

Blind Passage

A

Pass cells and fluid to a SECOND cell culture tube

21
Q

Cytopathic Effect

A

Visual morphologic changes in cells associated with specific virus infection

22
Q

Objective for observing cytopathic effects

A

Inverted phase contrast microscope (objective stage)

23
Q

Adenovirus in HEp-2 medium

A

Large rounding aggregates

24
Q

Confirming Adenovirus infection

A

Fluorescent antibody

25
Q

RSV in HEp-2 medium

A

Syncytia of cells

26
Q

Cytomegalovirus in HDF medium

A

Very slow discrete small foci of rounded cells

27
Q

Varicella-Zoster in HDF medium

A

Very slow rounded, swollen, refractile cells

28
Q

Herpes simplex virus in HEp-2, HDF, A549, RK medium

A

Rapid rounded, swollen, refractile cells

29
Q

Enterovirus in PMK medium

A

Refractile angular or tear-shaped cells

30
Q

Rhinovirus in HDF and PMK medium

A

Labile, HDF: Refractile rounding of cells, PMK: Refractile angular or tear-shaped cells

31
Q

Quantitation of Cytopathic Effect Uninfected monolayer

A

Negative

32
Q

Quantitation of Cytopathic Effect: Atypical alteration of monolayer involving few cells

A

Equivocal (±)

33
Q

Quantitation of Cytopathic Effect 1-25% of monolayer has CPE

A

1+

34
Q

Quantitation of Cytopathic Effect 25-50% of monolayer has CPE

A

2+

35
Q

Quantitation of Cytopathic Effect 50-75% of monolayer has CPE

A

3+

36
Q

Quantitation of Cytopathic Effect 76-100% of monolayer has CPE

A

4+

37
Q

R-mix shell vial for respiratory viruses (influenza)

A

1 type of Vx specific Ab

38
Q

Rapid virus detection

A

1-2 days instead of 5-28 days

39
Q

Disadvantage of shell vial cell culture

A

Can ONLY detect 1 type of virus per shell vial

40
Q

Provides the most sensitive method and is usually the more preferred technique compared to other alternative testing methods

A

NAAT

41
Q

Preferred testing method for common viruses Coronavirus Enterovirus Parechovirus Herpes virus

A

NAAT

42
Q

Preferred testing method for common viruses Adenovirus Influenza virus Parainfluenza virus RSV Varicella-Zoster virus Rotavirus Norovirus/Norwalk

A

NAAT, Antigen test

43
Q

Preferred testing method for common viruses Arbovirus, Cytomegalovirus, EBV, Parvovirus B19

A

NAAT, Antibody test

44
Q

Preferred testing method for common viruses= Hepatitis B Hepatitis D, HIV

A

NAAT, Antibody, antigen testing

45
Q

Preferred testing method for common viruses= Filovirus (Ebola) Arenavirus (LCMV, Lassa virus) Hantavirus Hepatitis A Hepatitis C Hepatitis E HTLV Rabies virus

A

Antibody test