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Diagnostic skills
Diagnostic skills
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1
Gross evaluation and histopathology. What is NOT true?
Unimportant, unuseful diagnostic method
Fixation of necropsy samples in 10% neutral buffered formalin
Consultation of lab tech and material details
Case history+ lesion description + samples collected
2
transfer of proteins from gel surface to special mem. Tests for FIV, confirmatory, detects antibodies. High specificity
Hemagglutination-Inhibition
Immunodiffusion
IgM class specific assay
Western Blot
3
Rationale for specific diagnosis. What is true regarding individual animal or herd level health?
Diagnosis needed for screening against viruses in embryo, blood, AI
ALL of the above
Establish management plan (vax, anti-microbial treatment)
Certification freedom, specific diagnosis for sale, need specific diagnosis
Zoonosis diseases like rabies, influenza, Venezuela Equine Encephalitis
Ex: CF for EIA, bovine leukemia, herpes
4
Image
Papillomavirus
Influenza
Herpesvirus
Rotavirus
Adenovirus
5
Inclusion bodies
Diagnose all viral infections
Light or electron microscopy
Acidophilic or basophilic
Acidophilic only
Basophilic only
Diagnose some but not all viral infections
6
Image
Rotavirus
Adenovirus
Herpesvirus
Influenza
Papillomavirus
7
Intracytoplasmic basophilic
Negribodies in rabies
Adenovirus/ owl eyes in cytomegalovirus
Feline rhinotracheitis (herpes)
Bollinger bodies in fowl pox
8
Image
Herpesvirus
Papillomavirus
Influenza
Rotavirus
Adenovirus
9
Detect viral antigens. Viability not required. One infected cell to 105 depend on assay and reagents. Specific to virus
ImmunoEM (IEM)
Immunofluorescence
Immunoperoxidase
10
Requires min 105 oral particles. Sensitivity is great. 1 animal or many. Rapid. High sensitivity, lower specificity
Pair sampling
Serological tests
ELISA/ 1-antibody capture
DIVA
Snap ELISA
11
Rationale for specific diagnosis. What is true?
many clinical cases occur as disease complex (ex: BRD, BDD)
All of the above
New strain or new virus
Gross pathology, histopathology, unusual clinical signs may not be helpful
Virus in clinical samples may be precurosor rather than causeative
12
Plaque assay for quantification and infectivity of viruses
Most accurate of quantitative biological assays
developed 1915-1917 in initial studies on bacteriophage
Brand spankin new
Concentrate over time and culture
Dilute over time and culture
Most accurate of qualititative chemical assays
13
CPE assay pros and cons
Support various cell lines
Requires careful specimen handling
Detect multiple viruses, including unknown viruses by CPE
False negative possible
False negative impossible
Some viruses do not produce CPE
14
Detection/growth of viruses in cell culture: What is true regarding pros and cons
Important for vaccine manufacture
Necessary for new pathogens, many viruses in one sample
Slow, labor intensive, time variability
Detects subtle CPE changes
Sesnitive and specific
Not always feasible and requires confirmatory testing
15
Degeneration changes in Mongolian cells caused by virus. Swell, round, shrink, syncytium, cluster, comp destruction
Cytonecrotic effects
Cytopathic effects
Cytomegaly
Cytitis
16
Intranuclear basophilic
Adenovirus/ owl eyes in cytomegalovirus
Feline rhinotracheitis (herpes)
Negribodies in rabies
Bollinger bodies in fowl pox
17
Immunoperoxidase, select the false statement
Fixation at room temp, less than 24 hours
Tissue fixed in 4% buffered formaldehyde
Large bits of tissue should be used
Fixative 155-20 times vol of tissue (change after first hour)
18
Detection of viruses by EM (negative stain and Thin section) : PROS
Dual infections can be detected better than antigen based testing
Allows direct visualization of noe or more viruses
Morphology allows for identification of unknown viruses
Skilled labor needed
Low sensitivity
Detection of non-cultivable viruses
19
What is true regarding Immunoperoxidase?
Better cellular localization
Light microscope can’t be used
Requires less than 24 hours
Fast processing
Not similar to IF labeling with enzyme to give it a color
Higher signal output with long term storage
20
Rationale for specific diagnosis. What is true? Regarding state, country, international level
All of the above
Specific diagnosis in a state provides sound veterinary service
Implemented for eradication programs like Marek, pseudorabies, BVD, EIA
International level may require diseases to be reported to gov officials
Allows for surveillance (FMD, swine fever, fowl plague, foreign incursions)
21
Intranuclear acidophilic
Negribodies in rabies
Bollinger bodies in fowl pox
Feline rhinotracheitis (herpes)
Adenovirus/ owl eyes in cytomegalovirus
22
Antibodies at time of patient presentation compared to 2-4 w. Diagnosis of influenza. Screen herd for effect of VAX
Snap ELISA
Pair sampling
ELISA/ 1-antibody capture
Serological tests
DIVA
23
Detect prior infection by virus. Example = Antibody detection in EIA, Bovine leukemia, caprine arthritis encephalitis
Pair sampling
ELISA/ 1-antibody capture
DIVA
Snap ELISA
Serological tests
24
Virus specific abs. One of most used methods for diagnosis of viruses that agglutinate RBC. Hemagglutinins present
IgM class specific assay
Hemagglutination-Inhibition
Immunodiffusion
Western Blot
25
Detect response to viruses in acute & chronic phases. Acute: blood IgM titer = west Nile, WEE, EE. Chronic/vax = IgG
Pair sampling
DIVA
Serological tests
Snap ELISA
ELISA/ 1-antibody capture
26
Intracytoplasmic acidophilic
Feline rhinotracheitis (herpes)
Bollinger bodies in fowl pox
Negribodies in rabies
Adenovirus/ owl eyes in cytomegalovirus
27
Known positive antiserum placed in alternating wells. Serum from sick animal also placed in well
Western Blot
Hemagglutination-Inhibition
IgM class specific assay
Immunodiffusion
28
Old, used to be commonly used. Agar gel. Ag + Ab diffuse against each other forming a line of precipitation.
Immunodiffusion
IgM class specific assay
Hemagglutination-Inhibition
Western Blot
29
Image
Influenza
Papillomavirus
Rotavirus
Adenovirus
Herpesvirus
30
Hemagglutination inhibition test (HAI) true?
Constant vol of RBC are added
Serum serially diluted
Constant viral antigen removed to avoid Reaction
Serum serially diluted
Constant viral antigen conc added allowed to react
Constant vol of RBC removed
31
Inclusion bodies
Nuclear or cytoplasmic aggregates of stainable carbs
Represent sites of viral degredation
Nuclear or cytoplasmic aggregates of stainable proteins (RNA, protein)
Represent sites of viral multiplication
32
SNT / Virus neutralization test is used on
TEquine Rhinopneumonitis, BVD antibodies
ALL of the above
Tick borne encephalitisi,
Avian infectious bronchitis, infectious bovine rhinotracheitis
Rabies vaccine efficiency
test vax efficiency and recent infection
33
Commercial test used for single animal. Simultaneously detect FeLV ag and FIV Ab
Pair sampling
Serological tests
Snap ELISA
DIVA
ELISA/ 1-antibody capture
34
Image
Rotavirus
Herpesvirus
Papillomavirus
Adenovirus
Influenza
35
Distinguish Abs response as result of natural infection vs vax. Avian influenza (deletion vax) and IBR (marker vax)
DIVA
ELISA/ 1-antibody capture
Snap ELISA
Pair sampling
Serological tests
36
Which are agent independent tests. For unknown viruses. Tell shape , size, componenents
PCR
E.M.
Cell culture
Elisa
IF
37
Virus neutralization test
Slow and labor intensive
Quick and easy, like me.
Species independent
Species dependent
38
Which are tests used to diagnose viral infections?
Seroconversion (antibodies)
Direct presence of infectious virus (antigens or nucleic acids)
All of the above
Direct visualization
39
Detection of viruses by EM (negative stain and Thin section) : CONS/ limitations
Concentration of virus in samples (10^6/ mL)
Allows direct visualization of noe or more viruses
Dual infections can be detected better than antigen based testing
Low sensitivity
Expensive
Skilled labor needed
40
Virus proliferation in cell cultures is detected following what?
Ultrasound
Radiograph
Electron imaging
Microscopic exam (10X)
41
Immunofluorescence, Click true statements
High sensitivity but lower specificity than ELISA
Slow results outside of 3 hours
Virus localization is a pro
Special equipment needed
Limited for certain viruses. Ex: group of enteroviruses
42
Detect viruses. More sensitive. Aggregation of virus in samples. Useful when virus is in low concentration
Immunoperoxidase
Iimmunofluorescence
ImmunoEM (IEM)
43
There are two methods to isolation and ID of viruses. Traditional & rapid. Which answer is filled with ONLY rapid tests?
ELISA, PCR, Necropsy
Isolation of virus in eggs and growth and isolatino in tissue cultures
ELISA, PCR
Histopathology and gross pathology in dead animals
44
Which are agent-dependent tests, specify known viruses
IF
Elisa
Cell culture
PCR
E.M.
45
ELISA, Virus neutralization assay, western blot, hemagluttination inhibition, Immunodiffusion, IgM class all assay what
Macrophages
Antigens
Neutrophils
Antibodies
