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MYCV > midterm examSPECIMEN CULTIVATION FOR VIROLOGY > Flashcards

midterm examSPECIMEN CULTIVATION FOR VIROLOGY Flashcards

1
Q

The primary purposes of viral cultivation are

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
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2
Q

The earliest method for the cultivation of viruses causing
human diseases was inoculation into __

A

human volunteers.

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3
Q

they used human volunteers for
their pioneering work on yellow fever

A

Reed and colleagues (1900)

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4
Q

Due to serious risk involved, human volunteers are used
only when __

A

when no other method is available and when the virus
is relatively harmless

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5
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

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6
Q

METHODS OF VIRAL ISOLATION

A
  1. Animal inoculation
  2. embryonated egg inoculation
  3. tissue culture
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7
Q

this type of isolation is the primary isolation of certain viruses

A

animal inoculation

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8
Q

this type of isolation is for us to study the pathogenesis, immune response, and epidemiology of a viral disease

A

animal inoculation

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9
Q

this isolation method as well is used for the study of oncogenesis

A

animal inoculation

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10
Q
  • they play
    an essential role in studies
    of viral pathogenesis
A

Laboratory animals

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11
Q

they used Monkeys for the
isolation of poliovirus

A

Landsteiner and Popper
(1909)

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12
Q

Landsteiner and Popper
(1909)-Monkeys for the
isolation of poliovirus

why is it risky?

A

potential to physical aggression and cause harm

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13
Q

he introduce white mice

A

Theiler (1903)

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14
Q

why monkey is not used anymore as a disease model?

A

they can’t exhibit human diseases

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15
Q

what type of mice is used in lab

A

Infant (Suckling mice)

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16
Q

suckling mice are susceptible to which viruses

A

Coxsackie and arbovirus

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17
Q

Routes of inoculation for animal inoculation

A

Intracerebral,
subcutaneous,
intraperitoneal,
intranasal

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18
Q

how do we confirm if there’s growth of virus inside the animal model

A

sign of Death, disease or visible
lesions

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19
Q

Disadvantages of animal inoculation

A
  1. costly
  2. maintenance
  3. interference of immune system
  4. individual variations (gender)
  5. difficulty in choosing of animals for particular virus
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20
Q

embryonated egg inoculation is started by whom

A

goodpasture (1931)

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21
Q

embryonated egg inoculation is started by goodpasture and is further developed by __

A

burnet

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22
Q

the embryonate egg we must use for inoculation is how many days old?

A

8-11 days old

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23
Q

the embryonated egg must be incubated for how many days

A

2-9 days

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24
Q
  • Eggs provide a suitable
    means for:
A

the primary isolation and identification of viruses

maintenance of stock cultures

and the production of vaccines

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25
Q

Routes of Inoculation of embryonated egg inoculation

A
  1. Chorioallantoic membrane(CAM)
  2. Amniotic Cavity
  3. Allantoic Cavity
  4. Yolk sac
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26
Q

in embryonated egg, viruses lies in Chorioallantoic membrane(CAM)

A

Herpes simplex virus
poxvirus
rous sarcoma virus

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27
Q

in embryonated egg, viruses lies in amniotic cavity

A

influenza virus
mumps virus

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28
Q

in embryonated egg, viruses lies in allantoic cavity

A

herpes simplex virus

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29
Q

in embryonated egg, viruses lies in yolk sac

A

influenza virus
mumps virus
newcastle disease virus
avian adenovirus

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30
Q

we look for ___ in Chorioallantoic Membrane
(CAM) to know if there’s growth of virus

A

pocks - visible lesion

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31
Q

most popular inoculation for embryonated egg

A

allantoic cavity

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32
Q

Has been widely used in
veterinary virology when it comes to embryonated egg

A

. Chorioallantoic Membrane
(CAM)

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33
Q

how do we interpret the result if it’s inoculated in allantoic cavity

A

fluid is examined for turbidity or hemagglutination

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34
Q

this type of inoculation for embryonated egg Has little application in
veterinary virology

A

amniotic cavity

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35
Q

Virus is introduced directly into
the amniotic fluid that bathes the
developing embryo

A

amniotic cavity

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36
Q

Volume of fluid in the
infected amniotic sac is small

A

(1-2 ml)

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37
Q

Simplest method for growth
and multiplication of virus in terms of embryonated egg

A

yolk sac

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38
Q

EMBRYONATED EGG ROUTES OF INOCULATION

Immune interference
mechanism can be detected
in most avian viruses

A

yolk sac

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39
Q

this inoculation method for embryonated egg Can also be used for the
cultivation of Chlamydia
and Rickettsia

A

yolk sac

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40
Q

Process of holding a strong light above
or below the egg to observe the
embryo

A

egg candling

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41
Q

Detection of Viral Growth in embryonated egg inoculation

A

death of the embryo
defect in embryonic development
localized areas of damage in the membranes (pocks)

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42
Q

A crucial technique in viral
isolation that involved
cultivating viruses in living
cells or tissues.

A

tissue culture

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43
Q

Provides a controlled
environment for studying
replication, pathogenesis
and development of
antiviral drugs and vaccines

A

tissue culture

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44
Q

TYPES OF TISSUE CULTURES

A

organ culture
explant culture
cell culture

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45
Q

a type of tissue culture wherein Small bits of organs can
be maintained in vitro

A

organ culture

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46
Q

a type of tissue culture wherein it’s useful for the isolation of some viruses which appear to be highly specialized parasites of certain organs

A

organ culture

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47
Q

a tissue culture wherein Fragments of minced
tissue can be grown as
‘explant’ embedded in
plasma clots

A

explant culture

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48
Q

a Routinely used type of tissue culture

A

cell culture

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49
Q

a type of tissue culture that is Dissociated using proteolytic
enzymes (trypsin) and
mechanical shaking

A

cell culture

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50
Q

the type of tissue culture in which the Growth medium contains essential amino acids,
vitamins, salts, glucose and a buffering system of bicarbonate in equilibrium
with atmosphere
containing 5% carbon
dioxide

A

cell culture

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51
Q

tissue culture specifically the cell culture, is Supplemented with up to
______ serum

A

5% calf or fetal calf

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52
Q

indicator used in cell culture

A

phenol red

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53
Q

CLASSIFICATION OF CELL CULTURES

A
  1. Primary Cell Cultures
  2. Diploid Cell Lines
  3. Continuous Cell Lines
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54
Q

the CLASSIFICATION OF CELL CULTURES is based on

A

origin
chromosomal characteristics
number of generations through which they can be maintained

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55
Q

this type of cell culture in which the Normal cells obtained from
fresh organs of animals or
human being and cultured

A

primary cell cultures

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56
Q

this type of cell culture is Capable of only limited growth
in culture and cannot be
maintained in serial culture (1-2
passage

A

primary cell culture

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57
Q

example of primary cell culture

A
  • Monkey kidney cell culture.
  • Human embryonic kidney.
  • Chick embryo cell culture
58
Q

this type of cell culture is Commonly employed for
primary isolation of viruses and
in preparation of vaccine

A

primary cell culture

59
Q

a type of cell culture that is a Diploid cells and have complete
set of chromosomes

A

diploid cell lines

60
Q

the type of cell culture - diploid cell lines has a * Limited lifespan ___

A

(20-50
serial passages)

61
Q

Useful for isolation of some
FASTIDIOUS pathogens and
for the production of viral
vaccines

A

diploid cell lines

62
Q

this type of cell culture which Cells of a single type, usually
derived from cancer cells, that are
capable of continuous serial
cultivation indefinitely

A

continuous cell lines

63
Q

example of continuous cell lines

A

Hela, hep-2 and KB cell lines

64
Q

continuous cell lines are Maintained by __

A

serial subcultivation
or stored in the cold (–70°C)

65
Q

is continuous cell line Now permitted to be used for
vaccine manufacture,?

A

yes, eg. rabies vaxx

66
Q

HeLa came from

A

Henrietta Lacks

67
Q

Hep 2 came from

A

human larynx epidermal carcinoma.

68
Q

Rapid modification of conventional
cell culture

A

shell vial cell culture

69
Q

shell vial cell culture

involved culturing cells in a small,
round-bottomed vial referred to
as a “__”

A

shell vial

70
Q

shell vial cell culture is Incubated for__hours

A

24 to 48

71
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
72
Q

one of the way to detect virus growth in a cell culture

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

A

viral inclusions

73
Q

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

A

syncytial cells

74
Q

Morphological changes in cultured cells - ‘___’ (CPE) caused by
‘cytopathogenic viruses.’

A

cytopathic effects

75
Q

Presumptive identification of a virus isolated from a clinical specimen.

A

cytopathic effect

76
Q

Main Types of CPE

A

rounding of cells
cell necrosis and lysis
syncytium formation
discrete focal degeneration
rounding and aggregation

77
Q

this virus causes a cytopathic effect such as rounding of cells

A

picornaviruses

78
Q

this virus causes a cytopathic effect such as cell necrosis and lysis

A

enteroviruses

79
Q

this virus causes a cytopathic effect such as syncytium formation

A

measles, respiratory syncytial virus, human immunodeficiency virus

80
Q

this virus causes a cytopathic effect such as discrete focal degeneration

A

herpes virus

81
Q

this virus causes a cytopathic effect such as rounding and aggregation

A

adenovirus

82
Q

quantitation of cell culture

negative

A

uninfected monolayer

83
Q

quantitation of cell culture

equivocal

A

atypical alteration of monolayer involving few cells

84
Q

quantitation of cell culture

1+

A

1%-25% of monolayer exhibits cytopathic effects

85
Q

quantitation of cell culture

2+

A

25%-50% of monolayer exhibits CPE

86
Q

quantitation of cell culture

3+

A

50%-75% of monolayer exhibits CPE

87
Q

quantitation of cell culture

4+

A

76% to 100 % of monolayer exhibits CPE

88
Q

an indicator of growth of virus in cell culture

Viruses interfere with the metabolic activities of infected cells
leading to reduction in cellular metabolic process

A

metabolic inhibition

89
Q

in metabolic inhibition, there will be changes in what parameter

A

pH

90
Q

in metabolic inhibition, normal cell culture’s medium will turn to what pH

A

medium turns basic because the normal cell culture must be acidic. If there’s a growth of virus, the medium will be basic

91
Q

virus in cell culture means ____ acid production in medium

A

no acid production causing the medium turn basic

92
Q

this indicator of growth of virus refers to a process wherein viruses binds to RBC

A

hemadsorption

93
Q

Viral envelope proteins may bind glycoproteins expressed on the surface of
erythrocytes
Addition of

A

hemadsorption

94
Q

hemadsorption

Addition of __ to the cultures

A

guinea pig erythrocytes

95
Q

what virus shows HEMADSORPTION as its indicator of cell culture growth

A

influenza and parainfluenza viruses

96
Q

Growth of the first will inhibit the infection of the second virus by
interference

A

interference

97
Q

Phenomenon for which a cell infected by a virus becomes resistant
toward a second outcoming infection by a superinfectant virus.

A

interference

98
Q

Non-cytopathogenic virus tested with known cytopathogenic virus

A

interference

99
Q

refers to a process and an indicator of cell culture growth predominantly practiced by Tumor forming (oncogenic) viruses

A

transformation

100
Q

Growth appears in a piled-up fashion
producing microtumors

A

transformation

101
Q

Tumor forming (oncogenic) viruses examples

A

herpes viruses
adenoviruses
hepadnavirus
papovavirues
retroviruses

102
Q

Technique used to visualize and localize specific antigens (viral
proteins) within cells.

A

immunofluorescence

103
Q

Gives positive results earlier than other methods

A

immunofluorescence

104
Q

stain used in immunofluorescence

A

Fluorescein Isothiocyanate (FITC)

105
Q

2 types of immunofluorescence

A

direct and indirect

106
Q

this type of immunofluorescence use A single labeled antibody is used to
directly detect the target viral antigen

A

direct immunofluorescence

107
Q

this type of immunofluorescence in which A primary antibody is followed by a
labeled secondary antibody, amplifying
the signal and increasing sensitivity

A

indirect immunofluorescence

108
Q

a more rapid but less sensitive type of immunofluorescence

A

direct IMF

109
Q

this type of immunofluorescence is best suited to large quantities of virus are suspected or when high quality, concentrated monoclonal antibodies are used

A

direct IMF

110
Q

used when lower quantities of virus are
suspected, such as detection of
respiratory viruses in specimens from
adult patient

A

indirect IMF

111
Q

Molecular-based assays example

A

polymerase chain reaction

112
Q

Provide rapid, sensitive, and specific methods of detection

A

DETECTION OF VIRUS-SPECIFIC NUCLEIC ACID

113
Q

Identified by detection of viral enzymes, such as reverse transcriptase in a virus ____ in the culture fluid

A

retroviruses

114
Q

Using antibodies to detect viral antigens or antibodies in the culture medium

A

detection of enzymes

115
Q

disadv of nucleic acid tsting

A

expensive
more specialized facilities needed
easily contaminated
low viral load but still have remnants will cause false positive

116
Q

Powerful technique used to visualize at a very high resolution of virus

A

electron microscope

117
Q

used to Study the structure, morphology and size of viral particles in d

A

electron microscope

118
Q

passes a beam of electrons through a thin specimen. The electron that pass
through interact with the specimen, creating an image

A

Transmission Electron Microscopy (TEM)

119
Q

Useful in studying the internal structure of viruses. It can reveal presence of viral
capsids, nucleic acid and other internal components

A

Transmission Electron Microscopy (TEM)

120
Q

2 VIRAL ASSAY

A

total virus particles
infectious virion assay

121
Q

2 methods under Total virus particles

A

electron microscopy
hemagglutination

122
Q

2 methods under infectious virion assay

A

quantal assays
quantitative infectivity assay

123
Q

Step 1 in electron microscopy

A

Negative Staining-virus suspension is mixed with a negative stain

124
Q

Step 2 in electron microscopy

A

: Latex Particle Addition-a known concentration of latex particles of a specific size is
added to the virus suspension. The latex particles serve as a visual reference

125
Q

Step 3 in electron microscopy

A

3: Electron Microscopy-The sample is examined under an electron microscope. The virus
particles and latex particles can be distinguished based on their size and
appearance

126
Q

Step 4 in electron microscopy

A

Counting-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

127
Q

A convenient method of quantitation for certain viruses particularly that possess
hemagglutinin proteins

A

Hemagglutination

128
Q

Hemagglutination is not a very sensitive indicator of the presence of small amount of
virus particles.

true or false

A

true

129
Q

steps in Hemagglutination

A

Step 1: Virus Dilution-Virus suspension is serially diluted
Step 2: Red blood cell addition-fixed volume of red blood cell is added to each dilution
incubate at 37°C`
Step 3: Observation of hemagglutination-if virus concentration is high enough, red blood
cells will clump together
Step 4: Endpoint Determination-Highest dilution that still produces hemagglutination is
determined

130
Q

Used to measure the “all-or-none” response of a population of virus

A

quantal assay

131
Q

Only indicates presence or absence of infectious viruses

A

quantal assay

132
Q

Quantal assays of infectivity can be carried out in __ for those
viruses

A

animals, eggs or tissue culture

133
Q

The virus sample is serially diluted to determine the lowest concentration that can still
produce infection

A

quantal assay

134
Q

Destruction of the host cell, embryo or animal or the appearance of CPE in cell cultures

A

quantal assay

135
Q

The titer of the original virus suspension is expressed as the 50% infectious
dose (ID50) or the 50% lethal dose (LD50) per milliliter

A

ID50 or LD50:

136
Q

measure the actual number of infectious
particles in the inoculum

A

Quantitative assays

137
Q

quantitative infectivity assay

Two methods are available—

A

plaque assay in monolayer cell culture
and pock assay on chick embryo CAM

138
Q

Viral suspension is added to a monolayer of cultured cells in a bottle or petri dish allowing time for
absorption

A

plaque assay

139
Q

Each infectious viral particle gives rise to a localized
focus of infected cells that can be seen with the naked
eye. - Known as ‘plaques’ and each plaque indicates
an infectious virus

A

plaque assay

140
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 solid agar gel to
prevent virus spreading throughout the culture

A

plaque assay

141
Q

___, form pocks when
inoculated onto the chorioallantoic
membrane of an embryonated egg.

A

Herpes and vaccinia

142
Q

in this assay, viruses can be assayed by
counting the number of pocks formed on
cam by appropriate inocula of virus

A

pock assay

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