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Serology > The Clinical Virology Laboratory > Flashcards

The Clinical Virology Laboratory Flashcards

1
Q

Specimen Collection:
- Samples may be collected from the site of _____ _____ or from a site where the virus is being ______ (asymptomatically)

A

viral pathology

shed

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2
Q

When should specimen be collected after onset of symptoms?

A

as soon as possible after onset of symptoms

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3
Q

What is the viral transport medium used for?

A

Used to prevent drying, maintain viability of virus, inhibit growth of contaminants

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4
Q

What specimen are placed in transport medium and what specimens are not?

A

Are: specimen from contaminated body sites

Are not: specimen from sites that are free from usual flora

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5
Q

Controls pH and keeps sample moist; prevents drying

A

Hanks Balanced salt solution or buffered broth

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6
Q

stabilizes or “cushions” virus; use fetal bovine serum, albumin, and gelatin

A

Proteinaceous Supplement

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7
Q

inhibits growth of bacteria and fungi

A

Antibiotics

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8
Q

signals the pH and indicates if pH is “off”; red/orange color change

A

pH indicator

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9
Q

collect in a sterile screw-capped container; do NOT use transport medium

A

spinal fluid

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10
Q

Rub on surface to collect cells and place in transport medium

A

Swab collection

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11
Q

collect in a sterile container; do NOT use transport medium

A

Urine collection

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12
Q

collect in anticoagulant such as heparin, EDTA, or sodium citrate; do NOT use transport medium

A

whole blood collection

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13
Q

If a sample will be processed within 3-5 days use ________ to store and ship on _____ ice.

A

refrigerator; wet

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14
Q

If sample will be processed after 3-5 days, use ________ at ____‘C or lower. Ship on ______ ice.

A

freezer; -70’C; dry

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15
Q

If respiratory syncytial virus (RSV) or cytomegalovirus (CMV) is suspected what don’t you do?

A

DO NOT Freeze

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16
Q

Two general steps in processing most clinical samples for viral culture

A

centrifugation and addition of antibiotics

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17
Q

following centrifugation of transport medium, the _______ is used to inoculate the cell culture tubes

A

supernatant

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18
Q

following centrifugation of transport medium, the _______ is used to prepare antigen smears for immunofluorecence testing

A

sediment

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19
Q

In whole blood samples, the ________ are separted out and used to inoculate cell cultures

A

leukocytes

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20
Q

In urine samples, _____-_______ urine is used to inoculate cell cultures

A

well-mixed

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21
Q

cell culture freshly explanted from tissue of origin; may have indigenous contaminants; ex: rhesus monkey kidney

A

primary cell culture

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22
Q

Cell cultures that have diploid chromosome number; can be passed 50 times so can’t survive long term; ex: MRC-5, MRHF

A

Diploid (Normal)

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23
Q

Cell cultures that have heteroploid chromosome number; originate from cancer tissue; can be passed indefinitely; Ex: A549

A

Established (heteroploid)

24
Q

In cell culture tubes how do viruses grow?

A

cells grow in a single layer on one side of the tube called a “monolayer” and are on their side to ensure they are bathed in medium

25
Q

inoculation performed by adding processed fluid to the cell culture tube; 2/10-3/10 of fluid

A

standard inoculation

26
Q

Inoculation procedure performed where the cell culture medium is decanted from the tube, processed material is applied to the monolayer, incubated in a slant rack (1hr at 35’C), excess inoculum is removed and idscarded, and fresh cell culture medium is added

A

Adsorption inoculation

27
Q

Incubation temperature of inoculated cell culture tubes

A

36-37’C on rotating and stationary racks (to keep the specimen out of the lid)

28
Q

____ _______ media contains the same components as transport media but have added enrichments such as amino acids, vitamins, and other nutrients.

A

cell culture

29
Q

color of cell culture medium in healthy cell culture tube

A

peach and clear

30
Q

cell culture medium color in a tube that is contaminated with bacteria or fungi

A

bright yellow and turbid

31
Q

cell culture medium color in which the pH is too basic

A

bright cherry pink

32
Q

how are cell culture tubes examined under a light microscope?

A
  • 10x with reduced light
  • railroad track to keep media in bottom of tube
  • look for changes in cell morphology
33
Q

changes in the cell morphology of viruses including swelling, shrinking, and detachment from the vessel

A

cytopathogenic effect (CPE)

34
Q

When are cell culture tubes are routinely examined?

A

on alternate days for 14 days

35
Q

the only culture that is examined for only 7 days

A

Herpes Simplex Virus (HSV)

36
Q

When are CMV cultures examined?

A

30 days , slowest grower

37
Q

Viral identity is PREDICTED by what 4 factors?

A

1 characteristic appearance of CPE
2 Time to appearance of the CPE
3 Cell line in which the CPE appears
4 source of the clinical sample

38
Q

Viral identity is CONFIRMED by what 2 factors?

A

1 Virus-infected cells are reacted with antibodies of known specificity and when they bind, identity confirmed
2 most common confirmatory method is immunofluorescence

39
Q

Procedure that can help detect viruses by replacing cell culture medium with a dilute suspension of erythrocytes (guinea pig) and refrigerating in a slant rack at 4’C for 30 min

A

Hemadsorption

40
Q

Describe a positive HAD test

A

RBCs stick to monolayer

41
Q

Describe a negative HAD test

A

the erythrocytes do not stick to the monolayer and float free in the tube

42
Q

Shell vial system procedure

A
  • clinical sample placed directly on monolayer
  • centrifuge at low speed for 1 hour
  • add medium and incubate
  • decant and add fixative
  • add stain to coverslip in vial
  • coverslips removed and mounted on a slide
  • fluroescence microscope to examine
43
Q

Shell vials are not evaluated for ______

A

CPE

44
Q

Advantages of shell vial

A

quicker detection, shorter turnaround time, no CPE to read

45
Q

Disadvantages to shell vial system

A

tedious to read, virus-specific (rather than “open” system)

46
Q

WHy are shell vials flexible?

A

specificity can be changed by changing the type of cells on the monolayer and the monoclonal antibodies used

47
Q

Monolayers of cells that include two different types of cells grown together

A

co-cultivated cell lines

48
Q

What are some marketed names for co-cultivated cell lines?

A

R-Mix (Respiratory viruses), E-Mix (for enteric viruses), H&V-Mix (for HSV, CMV, and VZV)

49
Q

Advantages of co-cultivated cell lines?

A
  • Cell lines retain usual characteristics of susceptibility to viruses
  • virology labs do not need to stock as many types of cell cultures
  • same as shell vial advantages
50
Q

Disadvantages of co-cultivated cell lines?

A

same as shell vial; tedious to read, virus specific

51
Q

Explain the meaning behind the picomaviridae family

A

small RNA viruses

52
Q

have fecal/oral route of transmission, replicate in the bowel but produce pathology at sites outside the bowel, most grow in standard cell cultures, difficult to differentiate from each other and from rhinoviruses, molecular methods are best for detection, ESPECIALLY for CSF samples

A

enteroviruses

53
Q

replicates in the bowel, moves to CNS to produce pathology, recent change from oral to injection vaccine, extinct in Western Hemisphere

A

Poliovirus

54
Q

trasmission and replication in the bowel, pathology may be CNS but more often in the heart

A

coxsackievirus A and B

55
Q

transmission and replication same as polio, may be isolated when disease is not present “orphan”

A

echovirus also called parechoviruses

56
Q

What does E. C. H. O stand for?

A

E-Enteric
C-cytopathogenic
H-human
O-orphan

57
Q

member of Picomaviridae family but is NOT an enterovirus, proliferate in upper respiratory tract and causes common cold, diagnosed clinically

A

Rhinovirus

Serology (17 decks)

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