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MIDTERMS > SPECIMEN COLLECTION AND CULTIVATION OF VIRUSES > Flashcards

SPECIMEN COLLECTION AND CULTIVATION OF VIRUSES Flashcards

1
Q
  • Specimen collection depends on the ____, ____
  • In the requisition, the ____ and ____ should be included.
  • Must be collected as early as possible after the onset of symptomatic disease
    ____ -interfere with nucleic acid based tests, recovery of some enveloped viruses, and fluorescent-antibody test
A

SPECIMEN COLLECTION FOR VIROLOGY
- specific disease syndrome, viral agents
suspected
- specimen type
- suspected virus
- Calcium alginate swabs

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2
Q

SUCCESSFUL LABORATORY INVESTIGATIONS

A

✓Collection of adequate and appropriate specimens
✓Sufficient documentation
✓Biosafety and decontamination
✓Correct packaging
✓Rapid transport
✓Choice of a laboratory that can accurately perform the tests
✓Timely communication of results

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3
Q

Specialized medium used to transport and store viral samples

A

VIRAL TRANSPORT MEDIUM (VTM)

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4
Q

Buffering Agents

A

Phosphate-Buffered saline: pH stability
and osmotic balance
▪Hanks’ balanced salt solution (HBSS):
mixture of salts and nutrients

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5
Q

Stabilizers

A

Glycerol: preservation of viral particles
▪Protein Stabilizers: bovine serum albumin
to stabilize the viral particles

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6
Q

Antimicrobial Agents

A

Antibiotics: penicillin, streptomycin,
gentamicin
▪Antifungal agents: amphotericin B

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7
Q

Throat swabs
1
2
3

A

Enteroviruses , Adenoviruses, HSV

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8
Q

Nasopharyngeal swab or aspirate
1
2
3

A

RSV , Influenza, Parainfluenza

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9
Q

Nasal Specimen

A

Rhinovirus

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10
Q

-____ are superior to swabs
-Swabs should be made of __,___,___,__
-Often are contaminated with bacteria.
> Contaminants may be removed by concentrating the sample through
___

A

THROAT, NASOPHARYNGEAL SWAB OR ASPIRATE
- aspirate
-polyester, Dacron, or rayon with plastic oraluminum shafts
- centrifugation

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11
Q
  1. Hold tongue away with tongue depressor
  2. Locate areas of inflammation and
    exudate in ____, tonsillar region of throat behind uvula
  3. Avoid swabbing ___; do not touch tongue
  4. Rub area back and forth with cotton or ___ swab
A

THROAT SWAB : POSTERIOR PHARYNGEAL SWAB
- posterior pharynx
- soft palate
- Dacron

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12
Q

1 Tilt head backwards
2 Insert _____ into nostril and back
to nasopharynx
3 Leave in place a few seconds
4 Withdraw slowly; rotating motion

A

NASOPHARYNGEAL SWAB
- flexible fine-shafted polyester swab

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13
Q
  1. Tilt head slightly backward
  2. Instill ___of VTM /sterile normal
    saline into one nostril
  3. Use ____
  4. Insert ___ in nostril and
    aspirate the secretion gently by
    suction in each nostril
A

NASOPHARYNGEAL ASPIRATE
- 1-1.5 ml
-aspiration trap
- silicon catheter

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14
Q

Specimen for detecting viruses that infect the lower respiratory tract( ___ and___)
Should be centrifuged to remove contaminating materials
>Not necessary if sample are for ____ and ____

A

BRONCHIAL AND BRONCHOALVEOLAR WASHES
- (Influenza and Adenoviruses)
- antigen
- nucleic acid testing

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15
Q

RECTAL SWABS AND STOOL is Used to detect __,__,___

A
  • Rotavirus, enteric adenoviruses (serotypes 40 and 41), and enteroviruses.
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16
Q

preferable for rotavirus and enteric adenovirus testing.

A

Stool

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17
Q

___ - acceptable for detecting enteroviruses in patients suspected of having an enteroviral disease, such as ___

A

Rectal swabs
- aseptic meningitis

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18
Q

Due to bacterial contamination in ____, centrifugation, filtration or both are
necessary for cell cultures for the recovery of viral agents
Storage: ____

A

stool
- 4°C or -15°C (Ag detection and PCR)

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19
Q

Urine is for detection of __,__,__,__,__,__
Improved recovery: at least ___ from clean catch first morning urine
Interference of urine __ and ___ may affect viral replication
>Urine is centrifuged or filtered to remove contaminants and neutralizing pH with____

A
  • CMV, Mumps, Rubella and Measles Virus,
    Polyomavirus and Adenovirus
  • 10mL
  • pH and bacteria
  • 7.5% solution of sodium bicarbonate
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20
Q
  • Detection of HSV or VZV may require a ___ if PCR testing is not available
    > are prepared by carefully unroofing the
    vesicle.
A

Tzanck smear

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21
Q

___, ___,____ , and in rare cases ___ or ___ can be detected in vesicular lesions of the skin and mucous membranes.
>Once the vesicle has ____ , detection of the virus is difficult

A
  • Enteroviruses, HSV, VZV
  • CMV or pox viruses
  • ulcerated or crusted
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22
Q

STERILE BODY FLUIDS OTHER THAN BLOOD
-may contain ___, __,___,___,____
- Collected aseptically by the physician and sent to the laboratory forprocessing
-Must not be diluted with VTM-causes___
- Specimens contaminated with blood may inhibit viral cultivation due to presence of ___

A
  • CSF and pericardial and pleural fluids, Amniotic Fluid
  • enteroviruses, HSV, VZV, influenza viruses, or CMV
  • false negative results
  • antibodies
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23
Q

Amniotic Fluid
1
2
3
.Delay in processing: Storage at refrigerated temp for ___ or ___ if longer

A

congenital CMV, VZV and parvovirus B19
- 48hrs or -70°C

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24
Q

Viral culture of blood is used primarily to detect __; however, __,__,__,__ occasionally may be encountered

A

CMV
- HSV, VZV, enteroviruses and adenovirus

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25
Q

___ of anticoagulated blood collected in a whole blood tube is needed
- __,__,___ anticoagulated blood is acceptable for CMV detection

A

5-10 mL
- Heparinized, citrated, or ethylene diaminetetraacetic acid (EDTA)

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26
Q

__ and __ should be used for samples collected for nucleic acid testing, because other anticoagulants may interfere with the enzyme functions required for PCR amplification.
Serum may be used for ___ and ____

A

EDTA and Citrated blood
- serologic tests and nucleic acid assays

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27
Q

For BONE MARROW
__ or __ anticoagulants are acceptable for culture
__ and ___ for nucleic acid testing

A

Heparin or EDTA
EDTA and ACD

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28
Q

Useful for detecting viruses that commonly infect the lungs (___,___,__,__), brain (___), and gastrointestinal tract (__).
Collected during surgical procedures.
- ___ is preferred for nucleic acid assays, but __ and ___ may be used after removal of the paraffin (deparaffinization) and extraction

A

Tissue
- CMV, influenza virus, adenovirus, sin nombre virus
- HSV
- CMV
- Fresh TISSUE
- formalin-fixed and paraffin-embedded tissues

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29
Q

GENITAL SPECIMENS
Detection of __ and __
Must be place in appropriate VTM

A

HSV and HPV

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30
Q

should be used for ulcerations and placed in
appropriate viral transport media.

A

Genital swabs

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31
Q

may be collected using a swab or brush and
placed in viral transport media.
Some manufactured __ or ___ are appropriate for nucleic acid testing

A

Cervical specimens
- endocervical or liquid-based cytology devices

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32
Q

__ and ___ specimens may be needed to detect
antibody to specific viruses
- Acute specimens should be collected as soon as possible after the
appearance of symptoms

A

Acute and convalescent serum

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33
Q

is collected a minimum of 2 to 3 weeks after the acute specimen

A

Convalescent specimen

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34
Q

Appropriate specimen is ___ of serum collected by venipuncture.

A

3 to 5 mL

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35
Q

Throat and Nasopharynx

A

Respiratory:
1 Adenoviruses Y
2 Parainfluenza virus Y
3 Influenza virus W
4 Respiratory syncytial virus W
5 Metapneumovirus W
6 SARS coronavirus W

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36
Q

nasal:+++ in respiratory

A

Rhinoviruses y

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37
Q

(other:serum) in respiratory

A

Sin nombre virus sp, s

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38
Q

Dermatologic and mucous membrane vesicular
Throat and Nasopharynx
Stool
Vesicle fluid or scrapping

A

Entero virus S F

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39
Q

Dermatologic and mucous membrane vesicular
Vesicle fluid or scrapping

A

Herpes simplex virus Y
Moknkeypox Y

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40
Q

Dermatologic and mucous membrane vesicular
Throat and Nasopharynx
Vesicle fluid or scrapping

A

Varicella zoster virus Y

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41
Q

Dermatologic and mucous membrane Exanthematous
Throat and Nasopharynx
Stool

A

Enterovirus S

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42
Q

Dermatologic and mucous membrane Exanthematous
Throat and Nasopharynx
Urine
Serum

A

Measles Y

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43
Q

Dermatologic and mucous membrane Exanthematous
Urine
Serum

A

Rubella Y

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44
Q

Dermatologic and mucous membrane Exanthematous
Serum
Amniotic fluid

A

Parvoviirus Y

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45
Q

Dermatologic and mucous membrane Pustular/Nodular
Tissue

A

Molluscum contagiosum, orf Y

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46
Q

Dermatologic and mucous membrane Pustular/Nodular
Tissue/ cells, thin prep

A

Warts Y

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47
Q

Dermatologic and mucous membrane Pustular/Nodular
cervical

A

Papillomavirus

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48
Q

Meningoencephalitis/encephalitis
CSF and Serum

A

Arboviruses S, F

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49
Q

Meningoencephalitis/encephalitis
Throat and Nasopharynx
CSF
Urine

A

Enteroviruses S, F

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50
Q

Meningoencephalitis/encephalitis
CSF
Brain biopsy

A

Herpes simplex virus Y

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51
Q

Meningoencephalitis/encephalitis
Serum

A

Lymphocytic choriomeningitis Y
Mumps virus Y

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52
Q

Meningoencephalitis/encephalitis
Brain biopsy

A

HIV Y
Polyomavirus (JC virus) Y

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53
Q

Meningoencephalitis/encephalitis
Corneal cels, brain

A

Rabies virus

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54
Q

Gastrointestinal dse
Stool

A

Adenoviruses (serotypes 40-41) Y
Noroviruses S
Rotavirus W, SP

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55
Q

Myocarditis, Pericarditis, and Pleurodynia
Throat and nasopharynx
csf
pericardial fluid

A

CoxsackieB S, F

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56
Q

Hemorrhagic fevers
Tissue, respiratory secretions and serum

A

Ebola/Malburg viruses Y

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57
Q

Hemorrhagic fevers
Throat and nasopharynx
URINE
Throat washes, serum

A

Lassa fever virus

58
Q

Hepatitis
blood

A

Hepatitis Y

59
Q

Dermatologic and Mucous membrane
Congenital and Perinatal
Throat/Nasopharynx
CSF
Urine

A

Enterovirus S

60
Q

Dermatologic and Mucous membrane
Congenital and Perinatal
Vesicle fluid

A

Herpes simplex virus Y

61
Q

Dermatologic and Mucous membrane
Congenital and Perinatal
Amniotic fluid, liver tissue

A

Parvovirus Y

62
Q

Dermatologic and Mucous membrane
Congenital and Perinatal
Urine
serum

A

CMV Y
Rubella Y
Zika Y

63
Q

Dermatologic and Mucous membrane
Eye (Ocular dse)
Throat and nasopharynx
conjunctival swab or scraping

A

Adenoviruses Y

64
Q

Dermatologic and Mucous membrane
Eye (Ocular dse)
conjunctival swab or scraping

A

Herpes simplex virus Y
Varicella- zoster virus Y

65
Q

Dermatologic and Mucous membrane
Post transplantation syndrome
urine
blood
tissue

A

Cytomegalovirus Y

66
Q

Dermatologic and Mucous membrane
Post transplantation syndrome
Blood tissue

A

Epstein barr virus Y

67
Q

Dermatologic and Mucous membrane
Post transplantation syndrome
BLOOD

A

Human herpesvirus-6 Y

68
Q

Dermatologic and Mucous membrane
Post transplantation syndrome
Tissue

A

herpes simplex Y

69
Q

Since the viruses are obligate intracellular parasites, they cannot be grown on any inanimate culture medium
Viruses can be cultivated within suitable hosts, such as a___

A

living celL

70
Q

The primary purposes of viral cultivation are:
1
2
3

A
  1. To isolate and identify viruses in clinical specimens
  2. To prepare viruses for vaccines
  3. And to do detailed research on viral structure, multiplication
    cycles, genetics, and effects on host cells
71
Q

The earliest method for the cultivation of viruses causing human diseases was inoculation into human volunteers.
>___ and ___ used human volunteers for
their pioneering work on yellow fever.
- Due to serious risk involved, human volunteers are used only when no other method is available and when the virus
is relatively harmless

A

Reed and colleagues (1900)

72
Q

LABORATORY DIAGNOSIS OF VIRAL INFECTION

A

I. Identification of the virus in cell culture
II. Microscopic identification in the specimen
III. Serological procedures to detect a rise in antibody titer
IV. Detection of viral antigen in blood or body fluids
V. Detection of viral nucleic acids

73
Q

METHODS OF VIRAL ISOLATION

A
  1. Animal Inoculation
  2. Embryonated Egg Inoculation
  3. Tissue Culture
74
Q

Primary isolation of certain viruses
* For the study of pathogenesis, immune
response and epidemiology of viral
diseases
* For the study oF ___

A

Animal Inoculation
- oncogenesis

75
Q

Laboratory animals play an essential role in studies of viral pathogenesis
* __ AND ___ -Monkeys for the isolation of poliovirus
* ____ -white mice
* monkeys, mice, rabbits,
guinea pigs, ferrets

A

Animal Inoculation
- Landsteiner and Popper (1909)
- Theiler (1903)

76
Q

limited application in virology

A

Monkey

77
Q

Infant:
Routes of inoculation of Animal inoculation: ____,___,___,___
Death, disease or visible lesions

A

Suckling mice
Coxsackie and arbovirus
Intracerebral, subcutaneous,
intraperitoneal, intranasal

78
Q

Animal Inoculation Disadvantages
1
2
3
4
5

A
  1. Costly
  2. Maintenance
  3. Interference of immune
    system
  4. Individual variations
  5. Difficulty in choosing of
    animals for particular
    virus
79
Q

____, further developed by Burnet
* 8-11 days old
* Incubated for 2-9 days

A

Embryonated Egg Inoculation
Goodpasture (1931)

80
Q

Eggs provide a suitable means for:
1
2
3

A
  • the primary isolation and identification of
    viruses
  • the maintenance of stock cultures
  • and the production of vaccines
81
Q

Embryonated Egg Inoculation
Routes of Inoculation
1
2
3
4

A
  1. Chorioallantoic membrane(CAM)
  2. Amniotic Cavity
  3. Allantoic Cavity
  4. Yolk sac
82
Q

Has been widely used in veterinary virology
* Viruses grow readily or can be adapted to grow on it
* Produces visible lesions ____
* ____, can be used for the assay of pock- forming viruses
* Different viruses have different pock morphology

A

Chorioallantoic Membrane
(CAM)
- (pocks)
- Pock counting

83
Q

Most popular
* Provides a rich yield of influenza
and some paramyxoviruses for
vaccine production
* ___,__, __,___
* Fluid is examined for ___ or
____

A

Allantoic Cavity
- Influenza Virus, Mumps Virus,
Newcastle disease virus, Avian
Adenovirus
- turbidity or hemagglutination

84
Q

Virus is introduced directly into it, that bathes the developing embryo
* Volume of fluid in the infected____
* Recommended for the primary isolation of human viruses:
1
2
* Has little application in veterinary virology
* Newly isolated influenza viruses may require several passages before they adapt to growth by other routes, such as ___

A

Amniotic Cavity
- amniotic sac is small (1-2 ml)
-Mumps virus
*Influenza A, B and C viruses
- allantoic

85
Q

Simplest method for growth
and multiplication of virus
- __,__,__,__
* Immune interference mechanism can be detected in most __ viruses
* Can also be used for the
cultivation of __, and __

A

Yolk Sac
- * Influenza, HSV, Vaccinia
and some Arboviruses
- avian
-Chlamydia and Rickettsia

86
Q

Process of holding a strong light above or below the egg to observe the embryo
A candling lamp consists of a strong
electric bulb covered by a plastic or
aluminum container that has a handle
and an aperture

A

Egg Candling

87
Q

Detection of Viral Growth
1
2
3

A
  • Death of the embryo
  • Defects in embryonic development
  • Localized areas of damage in the membranes
    (pocks)
88
Q

A crucial technique in viral isolation that involved cultivating viruses in living
cells or tissues.
- Provides a controlled environment for studying replication, pathogenesis
and development of antiviral drugs and vaccines

A

Tissue Culture

89
Q

TYPES OF TISSUE CULTURES
1
2
3

A
  1. Organ Culture
  2. Explant Culture
  3. Cell Culture
90
Q

Small bits of organs can be maintained in ___
* Useful for the isolation of some viruses which appear to be highly specialized parasites of certain organs

A

Organ Culture
- vitro

91
Q

Fragments of minced tissue can be grown as
‘explant’ embedded in ___
* _____ were used for the isolation of adenoviruses

A

Explant Culture
- plasma clots
- Adenoid tissue explant
culture

92
Q

Routinely used
* Dissociated using __ and ____
* Growth medium contains
__,__,__,___ and a buffering system of
____ in equilibrium with atmosphere
containing____

A

Cell Culture
- proteolytic enzymes (trypsin) and
mechanical shaking
-essential amino acids, vitamins, salts, glucose
-bicarbonate
- 5% carbon dioxide

93
Q

Supplemented with up to 5% calf or fetal calf serum
* ___ to prevent bacterial contamination
* ___ as indicator

A

Cell Culture
-Antibiotics
- Phenol red

94
Q

CLASSIFICATION OF CELL CULTURES
3 Types based on:

A

Origin
▪ Chromosomal characteristics
▪ Number of generations through which they can be maintained

95
Q

CLASSIFICATION OF CELL CULTURES
1
2
3

A
  1. Primary Cell Cultures
  2. Diploid Cell Lines
  3. Continuous Cell Lines
96
Q

Normal cells obtained from fresh organs of animals or human being and cultured.
* Capable of only limited growth in culture and cannot be maintained in serial culture (1-2
passages)
1
2
3
* Commonly employed for primary isolation of viruses and in preparation of vaccine

A

Primary Cell Cultures
- Monkey kidney cell culture.
- Human embryonic kidney.
- Chick embryo cell culture.

97
Q

have complete set of chromosomes
* Limited lifespan (___ serial passages)
* Useful for isolation of some ___ and for the production of viral vaccines

A

Diploid Cell Lines
20-50
fastidious pathogens

98
Q

Cells of a single type, usually derived from ____, that are capable of continuous serial cultivation indefinitely
*__,__ and ___have been used in lab
throughout the world for many years
* Maintained by serial subcultivation
or stored in ____
* Now permitted to be used for vaccine manufacture, for example, ____ for rabies vaccine

A

Continuous Cell Lines
- cancer cells
- Hela, hep-2 and KB cell lines
- cold (–70°C)
- vero cell

99
Q

Primary cell cultures

A

Rhesus monkey kidney cell structure
Human amnion cell culture
Chick embryo fibroblast cell culture

100
Q

Diploid cell strains

A

WI-38 (Human Embryonic lung cell strain)
HL-8 (Rhesus embryo cell strain)

101
Q

Continuous cell line

A

HeLA (Human carcinoma of cervix cell line)
HEP-2 (Human epithelioma of larynx cell line)
KB ( Human carcinoma nasopharynx cell line)
McCoy (Human synovial carcinoma cell line)
Detroit-6 (Sternal marrow cell line)
Chang C/I/L/K (Human conjunctiva (c), Intestine (I) lIVER (L) Kidney (K) cell lines)
Vero (Vervet monkey kidney cell line)
BHL 21 (Baby Hamster kidney cell line)

102
Q
  • Derived from freshly isolated tissues or organs
  • Normal, diploid (46chromosomes)
  • limited (1-5 passages)
    -Slow growth, contact inhibition
  • Undergo senescence quickly
A

Primary Cell Cultures

103
Q

Derived from normal, diploid cells
Normal, diploid (46 chromosomes in humans)
Finite (typically 20-50 population doublings)
Slow growth, contact inhibition
Undergo senescence after finite number of
passages

A

Diploid Cell Lines

104
Q

Derived from cancer cells or transformed
cells
Abnormal, aneuploid (variable number of
chromosomes)
Unlimited (can be maintained indefinitely)
Rapid growth, loss of contact inhibition

A

Continuous Cell Lines

105
Q
  • Rapid modification of conventional
    cell culture
  • Involved culturing cells in a small,
    round-bottomed vial
    Inoculated with the clinical sample
    and then centrifuged to promote
    viral absorption.
    Incubated for ___
A

SHELL VIAL CELL CULTURE
- 24 to 48 hours

106
Q

DETECTION OF VIRUS GROWTH IN CELL CULTURE

A
  1. Cytopathic effect
  2. Metabolic inhibition
  3. Hemadsorption
  4. Interference
  5. Transformation
  6. Immunofluorescence
  7. Detection of virus-specific nucleic acid
  8. Detection of enzymes
107
Q

components in an infected cell or abnormal
accumulations of cellular materials resulting from virus-induced metabolic disruption.

A

Viral inclusions

108
Q

-aggregates of cells fused to form one large cell
with multiple nuclei

A

Syncytial cells

109
Q

Presumptive identification of a virus isolated from a clinical specimen.
Morphological changes in cultured cells

A

CYTOPATHIC EFFECT

110
Q

Main Types of CPE
1
2
3
4
5

A
  1. Rounding of cells - picornaviruses
  2. Cell necrosis and lysis – Enteroviruses
  3. Syncytium formation - measles, respiratory syncytial virus, human
    immunodeficiency virus (HIV)
  4. Discrete focal degeneration - Herpes virus
  5. Rounding and aggregation - Adenovirus
111
Q

Quantification of cell cytopathic effects

A

Negative uninfected monolayer
Equivocal +/- Atypical alteration of monolayer invoviving few cells
1+ 1-25% of monolayer exhibits cpe
2+ 25-50% of monolayer exhibits cpe
3+ 50-75% of monolayer exhibits cpe
4+ 76-100% of monolayer exhibits cpe

112
Q

Viruses interfere with the metabolic activities of infected cells leading to reduction in cellular metabolic processes
Changes in ___ in the culture medium.
Normal cell cultures-medium turns __
Virus in cell culture-__

A

METABOLIC INHIBITION
- pH
- acidic
- no acid production

113
Q

Viral envelope proteins may bind glycoproteins expressed on the surface of erythrocytes
Addition of guinea pig erythrocytes to the cultures - __ and ____

A

HEMADSORPTION
- Influenza and parainfluenza viruses

114
Q

Non-cytopathogenic virus tested with known cytopathogenic virus
Growth of the first will inhibit the infection of the second virus by this
Phenomenon for which a cell infected by a virus becomes resistant toward a second outcoming infection by a superinfectant virus

A

INTERFERENCE

115
Q

Tumor forming (oncogenic) viruses
Growth appears in a piled-up fashion
producing microtumors
1
2
3
4
5

A

TRANSFORMATION
Herpes viruses
Adenoviruses
Hepadnavirus
Papovaviruses
Retroviruses

116
Q

Technique used to visualize and localize specific antigens (viral proteins) within cells.
Gives positive results earlier than other methods
___

A

IMMUNOFLUORESCENCE
- Fluorescein Isothiocyanate (FITC)

117
Q

A single labeled antibody is used to directly detect the target viral antigen
- More rapid but less sensitive
- best suited to large quantities of virus
are suspected or when high-quality,
concentrated monoclonal antibodies are
used

A

Direct Immunofluorescence

118
Q

A primary antibody is followed by a
labeled secondary antibody, amplifying
the signal and increasing sensitivity
used when lower quantities of virus are
suspected, such as detection of
respiratory viruses in specimens from
adult patients

A

Indirect Immunofluorescence

119
Q

Interpretation of Fluorescence intensity using fitc

A

Negative No apple green fluorescence
1+ Faint yet unequivocal apple green fluorescence
2+ Apple green fluorescence
3+ Bright apple green fluorescence
4+ Brilliant apple green fluorescence

120
Q

Molecular-based assays, such as polymerase chain reaction
Provide rapid, sensitive, and specific methods of detection

A

DETECTION OF VIRUS-SPECIFIC NUCLEIC ACID

121
Q

Identified as reverse transcriptase in ___ , in the culture fluid.
Using antibodies to detect viral antigens or antibodies in the culture medium

A

DETECTION OF ENZYMES
- retroviruses

122
Q

Powerful technique used to visualize at a very high resolution
Study the structure, morphology and size of viral particles in detail

A

ELECTRON MICROSCOPY

123
Q
  • passes a beam of electrons through a thin specimen. The electron that pass
    through interact with the specimen, creating an image
  • Useful in studying the internal structure of viruses. It can reveal presence of ____,___ and ____
A

Transmission Electron Microscopy (TEM)
- viral capsids, nucleic acid and other internal components

124
Q

VIRAL ASSAY

A
  1. Total virus particles
  2. Infectious virion assay
125
Q

Total virus particles

A

Electron microscopy
Hemagglutination

126
Q

Infectious virion assay

A

Quantal assays
Quantitative infectivity assay

127
Q

Direct virus counting with electron microscopy and latex particles

A

Electron Microscopy

128
Q

Step 1: ____ -virus suspension is mixed with a negative stain
Step 2:____-a known concentration of latex particles of a specific size is
added to the virus suspension. The latex particles serve as a ___
Step 3:___ -The sample is examined under an electron microscope. The virus
particles and latex particles can be distinguished based on their ___ and___
Step 4: ____ -virus particles and latex particles are counted in a specific area of the electron micrograph. The ratio between the two can be used to estimate the concentration of the virus particles in the original suspension

A

Electron Microscopy
- Negative Staining
- Latex Particle Addition
- visual reference
- Electron Microscopy
- size and appearance
- Counting

129
Q

Used to measure the “all-or-none” response of a population of virus.
Only indicates presence or absence of infectious viruses
can be carried out in animals, eggs or tissue culture for those viruses

A

Quantal Assays

130
Q

The virus sample is serially diluted to determine the lowest concentration that can still produce infection
Destruction of the host cell, embryo or animal or the appearance of CPE in cell cultures

A

Quantal Assays

131
Q

__ or __: The titer of the original virus suspension is expressed as the 50% infectious
dose or the 50% lethal dose per milliliter

A

ID50 or LD50

132
Q

A convenient method of quantitation for certain viruses particularly that possess
hemagglutinin proteins
- is not a very sensitive indicator of the presence of small amount of virus particles.
- approximately ____ are required to produce macroscopic agglutination.

A

Hemagglutination
- 107 influenza virions

133
Q

Step 1: ____-Virus suspension is serially diluted
Step 2: ____-fixed volume of red blood cell is added to each dilution
incubate at ___
Step 3:___ -if virus concentration is high enough, red blood
cells will clump together
Step 4: ___-Highest dilution that still produces hemagglutination is determined

A

Hemagglutination
- Virus Dilution
- Red blood cell addition
- 37°C
- Observation of hemagglutination
- Endpoint Determination

134
Q

measure the actual number of infectious
particles in the inoculum.
Two methods are available
1
2

A

Quantitative Infectivity Assay
- plaque assay in monolayer cell culture
and pock assay on chick embryo CAM

135
Q

Viral suspension is added to a monolayer of cultured cells in a bottle or petri dish allowing time for absorption,
- Removed and replaced with a ___ to prevent virus spreading throughout the culture
- Each infectious viral particle gives rise to a localized focus of infected cells that can be seen with the naked eye. -

A

PLAQUE ASSAY
- solid agar gel

136
Q

__ and ___, form pocks when inoculated onto the chorioallantoic membrane of an embryonated egg.
- Such viruses can be assayed by counting the number of pocks formed on cam by appropriate inocula of virus

A

POCK ASSAY
- Herpes and vaccinia

137
Q

Each plaque represents ____ in the original virus sample.

A

1 infectious unit (virion or infected cell)

138
Q

choriallantoic membrane inoculation

A

hsv
pox virus
rous sarcoma virus

139
Q

amniotic inoculation

A

influenza
mumps

140
Q

yolk sac inoculation

A

hsv

141
Q

allantoic inoculation

A

influenza
mumps
newcastle dse virus
avian adenovirus

MIDTERMS (19 decks)

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