SPECIMEN COLLECTION TO ADENOVIRIDAE Flashcards
(119 cards)
1
Q
[?], from the [?]
Taken at the [?]
A
RIGHT SPECIMEN; RIGHT SITE
RIGHT TIME
2
Q
Stored and transported [?]
A
IMMEDIATELY
3
Q
To ensure [?]
A
accurate laboratory results and prevent contamination.
4
Q
: most crucial procedure
A
Pre – analytical
5
Q
Store specimen at [?]
A
4C
6
Q
Transportation that will take days at [?]
A
-70C
7
Q
SPECIMENS
A
• Urine
• Throat Swab/Throat Washings
• Blood; Plasma
• Rectal swab
• Biopsy material
• Skin scrapings
• CSF
• Vesicle fluid
• Autopsy tissue
8
Q
Respiratory Infection
A
Nasal or throat swabs, postnasal washings
9
Q
Gastrointestinal Infection
A
Feces
10
Q
Vesicular Rash
A
Vesicular fluid
11
Q
Hepatitis
A
Serum, feces
12
Q
CNS
A
CSF, throat swab, feces
13
Q
AIDS
A
Unclotted blood
14
Q
VIRAL GROUP
A
Respiratory Tract
Mouth or Gastrointestinal Tract
Skin
15
Q
Respiratory Tract
A
• Adenoviridae
• Herpesviridae
• Poxviridae
• Picornaviridae
• Togaviridae
• Orthomyxovirid ae
16
Q
Mouth or Gastrointestinal Tract
A
• Adenoviridae
• Herpesviridae
• Picornaviridae
• Reoviridae
• Flaviviridae
17
Q
Skin: A. Mild Trauma
A
• Papovaviridae
• Herpesviridae
• Poxviridae
18
Q
Skin: B. Injection
A
• Herpesviridae
• Hepadnaviridae
• Retropiridae
19
Q
Skin: C. Bites
A
• Togaviridae
• Rhabdoviridae
• Flaviviridae
20
Q
THREE APPROACHES
A
Direct detection of virus
Virus Isolation (Indirect detection)
Serology (Detection of Antibodies)
21
Q
• Electron Microscopy
A
Direct detection of virus
22
Q
• Light Microscopy “Inclusion Bodies”
A
Direct detection of virus
23
Q
• Antigen detection tests
A
Direct detection of virus
24
Q
• Molecular methods: PCR (Polymerase Chain Reaction) & Nucleic Acid Probes
A
Direct detection of virus
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• Animal inoculation Inoculation of eggs
Virus Isolation (Indirect detection)
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• In vitro Cell Culture
Virus Isolation (Indirect detection)
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microscopic examination for viral inclusions
CYTOLOGY/HISTOLOGY
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less sensitive than culture
CYTOLOGY/HISTOLOGY
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helpful for viruses that are difficult to grow
CYTOLOGY/HISTOLOGY
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Cytology:
herpes simplex, varicella zoster
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Histology:
adenivirus, cytomegalovirus, measles, papilloma virus, parvovirus, rabies (infects the brain)
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ELECTRON MICROSCOPY
Advantages
• Economical running costs?
• Detects ?
• Adaptable for the confirmation of ?
• ? required for result; results available the same day (fastest)
• Specific ?
cost
unculturable viruses
cytopathic effect
Short length of time
antiviral chemotherapy
33
ELECTRON MICROSCOPY
Disadvantages
• Requires ?
• Poor ?
• Large ?
skilled staff
sensitivity
capital outlay
34
Examples of viruses seen: gastroenteritis
o Calici virus
o Corona virus
o Enteric virus
o Adeno virus
o Norwalk
35
Examples of viruses seen: encephalitis
o John Cunningham (JC) virus
o Herpes Simplex Virus
o Measles
36
TYPES OF ELECTRON MICROSCOPY
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IMMUNOFLUO RESCENT MICROSCOPY Examples:
Adenovirus, CMV, herpes simplex 1 and 2, influenza virus
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antiviral antibodies are used to stain viral antigens in specimen and cultures
IMMUNOFLUO RESCENT MICROSCOPY
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Black background
IMMUNOFLUO RESCENT MICROSCOPY
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usually solid-phase and membrane Elisa are used
ENZYME IMMUNOASSA Y (EIA)
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the basic test consists of antibodies bonded to enzymes, the enzymes remain able to catalyze a reaction yielding a visible discernible end product while attached to antibodies
ENZYME IMMUNOASSA Y (EIA)
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ENZYME IMMUNOASSA Y (EIA)
Examples:
enteric adenovirus, rotavirus, RSV
43
Notes :
• Sample : Stool
• Collecting specimens, have to poke or collect in 3 different sites (e.g. left, right)
• Right amount is placed in a solution → shake → place in a cartridge → results in 10 to 20 mins
ENZYME IMMUNOASSA Y (EIA)
44
baby hamster kidney cell culture with Bgalactosidase gene that is expressed when cells are infected with the virus
ENZYME LINKED VIRUS INDUCIBLE SYSTEM (ELVIS)
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ENZYME LINKED VIRUS INDUCIBLE SYSTEM (ELVIS)
Example:
Herpes simplex 1 and 2
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different viruses grow on different cell lines
CONVENTIONAL CELL CULTURE
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growth takes about 1 - 28 days
CONVENTIONAL CELL CULTURE
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microscopically examined for cytopathic effects (CPE)
CONVENTIONAL CELL CULTURE
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Cell rounding, granulation, clumping, vacuolation
cytopathic effects (CPE)
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Giant multinucleated cells formation: Herpes simplex virus, Tzanck smear
cytopathic effects (CPE)
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Cell fusion, lysis, flux, inclusion bodies
cytopathic effects (CPE)
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confirmed with immunofluorescent stains
CONVENTIONAL CELL CULTURE
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CONVENTIONAL CELL CULTURE
Examples:
Adenovirus, CMV, enteroviruses, RSV
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THREE BASIC TYPES OF CELL CULTURES
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made by dispersing cell (usually with trypsin) from freshly removed host tissues
PRIMARY CELL LINE
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Unable to grow for more than a few passages in culture
PRIMARY CELL LINE
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PRIMARY CELL LINE Example :
Primary Monkey Kidney
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that have undergone a change that allows their limited culture (range from 50 up to 100 passages/generations) but that retain their normal chromosome pattern
DIPLOID CELL LINE
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DIPLOID CELL LINE Example :
Human diploid fibroblast and Human diploid vaccine for Lassa virus/Rabies
60
cultures capable of more prolonged growth ("immortal") that have been derived from diploid cell lines or from malignant tissues
CONTINUOUS CELL LINE
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INTRACYTOPLASMIC INCLUSION BODIES
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INTRANUCLEAR INCLUSION BODIES
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BOTH INTRACYTOPLASMIC AND INTRANUCLEAR
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NEGRI BODIES
Rabies virus
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MOLLUSCUM BODIES
Molluscum contagiosum virus
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GUARNIERI BODIES
Vaccinia virus
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OWL'S EYE INCLUSION BODIES
Cytomegalovirus
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COWDRY TYPE A BODIES
HSV, VZV
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COWDRY TYPE B BODIES
Polio virus
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TORRES BODIES
Yellow Fever Virus
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WARTHIN-FINKELDEY BODIES
Measles virus
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• Grow via conventional cell culture (Cytomegalovirus and Varicella zoster virus)
SHELL VIAL CULTURE
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• Advantage: Rapid, within 1-2 days
SHELL VIAL CULTURE
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• Direct test, latex coated with antibodies, rapid and inexpensive
LATEX AGGLUTINATION
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• Example: Rotavirus
LATEX AGGLUTINATION
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• Used in IS
• Agglutination = (+)
LATEX AGGLUTINATION
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• Limited commercial availability
POLYMERASE CHAIN REACTION (PCR)
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• Enterovirus in CSF
POLYMERASE CHAIN REACTION (PCR)
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• Most common: used during the pandemic (confirmatory test for + result)
POLYMERASE CHAIN REACTION (PCR)
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• Specific IgG assays are good indications of prior infections
SEROLOGICAL TEST
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• IgM assays are good indicators of recent or recent infections
SEROLOGICAL TEST
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• Allow retrospective diagnosis if no acute clinical samples are available
SEROLOGICAL TEST
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• Rapid
SEROLOGICAL TEST
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• May be automated
SEROLOGICAL TEST
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• Allows diagnosis if uncultured viruses are involved
SEROLOGICAL TEST
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• Can make use of noninvasive clinical samples such as urine saliva
SEROLOGICAL TEST
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• CF test is usually insensitive
SEROLOGICAL TEST
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• Cross-reactivity of antibodies is a source of false positive results v Insensitive for congenital infections
SEROLOGICAL TEST
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• Not appropriate for immunocompromised patients
SEROLOGICAL TEST
90
SEROLOGICAL TEST Example : in Isolating the virus
Antigen detection test by Immunofluorescence
Complement fixation
Hemagglutination inhibition
Enzyme immunoassay
91
DETECTED IN SERUM ONLY
SEROLOGICAL TEST
92
Useful in evaluating immune status or diagnosing viral infections where culture is difficult or impossible
SEROLOGICAL TEST
93
SEROLOGICAL TEST Example virus :
Arbo virus
Cytomegalovirus
Epstein- Barr virus
Hepatitis
Herpes simplex
Measles
Rubella
Varicella zoster virus
94
Adenoviridae
a. Human Adenovirus
95
• Hepadnaviridae
a. Hepatitis B Virus
96
• Herpes viridae
a. Herpes Simplex Virus 1
b. Herpes Simplex Virus 2
c. Varicella-Zoster Virus
d. Cytomegalovirus
e. Epstein-Barr Virus
f. Human Herpes Virus 6,7 and 8
97
• Papovaviridae
a. Human Papilloma Virus
b. JC and BK Polyoma Virus
98
• Parvoviridae
a. Parvovirus B-19
99
• Poxviridae
a. Variola
b. Vaccina
c. Orf
d. Molluscum contagiosum
100
• Double-stranded (except)
Parvoviridae
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• Icosahedral (except)
Poxviridae
102
• Replication in the nucleus (except)
Poxviridae
103
• Naked:
Papoviridae, Parvoviridae, Adenoviridae
104
• Enveloped:
Poxviridae, Herpesviridae, Hepadnaviridae
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subspecie
Human Adenovirus
106
over 80 species with about 40 are pathogenic to man
Human Adenovirus
107
70-90 nm
Human Adenovirus
108
Isolated from humans and animals
Human Adenovirus
109
produces cytopathic changes
ADENOVIRIDAE
110
rounding, enlargement and aggregation
ADENOVIRIDAE
111
Diseases
• Acute Febrile Pharyngitis: common in young children
• Pharyngeal-conjunctival Fever
• Eye infections
• Pneumonia and gastroenteritis
112
- Common in newborn, immunosuppressed and military men
Eye infections
113
No ab produced
ADENOVIRIDAE
114
Cytotoxic T-cells kill infected cell early
ADENOVIRIDAE
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Common to children below 6 y/o
ADENOVIRIDAE
116
Predominant in cold climate
flu-like symptoms; not fatal
ADENOVIRIDAE
117
ADENOVIRIDAE SAMPLE COLLECTION
throat swab, eye samples, stool
118
ADENOVIRIDAE DIAGNOSIS
cell culture, serologic test and control
119
ADENOVIRIDAE CONTROL
vaccine; Anti-flu vaccine
