(Final) Diagnosis of viral infections Flashcards
What are the risks levels associated with broad risk categories of viral infections?
Risk Group 1 = No or low individual risk
Risk Group 2= Moderate individual risk, low community risk
Risk Group 3 = High individual risk, low community risk
Risk Group 4 = High individual risk, High community risk
What risk category is Foot and mouth disease classified as?
Risk group 4
T/F: BSL4 is the maximum containment laboratory. BSL4 labs handle dangerous and exotic pathogens belonging to the highest risk group - 4
TRUE
ex: ebola virus
What is a biohazard?
Biological substances that pose a threat to the health of living organisms, primarily that of humans
What is biosafety?
Laboratory biosafety describes the containment principles, technologies, and practices that are implemented to prevent unintentional exposure to pathogens and toxins, or their accidental release
Very small droplets of fluid that can spread via air are called?
Aerosol
Viruses can spread in the lab through aerosol route
What does biosecurity refer to?
Lab biosecurity described the protection, control, and accountability for valuable biological materials within labs - in order to prevent their unauthorized access, loss, theft, and misuse
When should samples be taken for virus isolation?
Specimens should be collected as soon after onset of symptoms as possible. *maximum amounts (titers) of virus are usually present at the onset of signs
**chance of viral recovery is best during the first three days after onset - up to five
When should samples be taken for viral serological tests?
Two blood samples are best: One during the acute phase of illness and the second sample during the convalescence period
When should samples be collected for PCR or molecular diagnostics?
During the early part of illness
Viral swab samples should be sent to the lab in what medium?
Viral transport medium (VTM)
To prevent spillage, it is recommended to follow the _______________ system while transporting infectious materials
Basic triple packing system
What would be a good time frame to collect samples for virus isolation?
- within 3 days of onset of clincial signs
- within 7 days of onset of clinical signs
- within 14 days of onset of clinical signs
- within one month of onset of clincal signs
1 - within 3 days of onset of clinical signs
What process must be done to tissues prior to virus testing?
Tissue homogenization
Via tissue grinder or mortar and pestle
What are three simple ways to diagnose the presence of a specific virus?
Gross evaluation and histopathology:
Clinical signs
Necropsy
Histopath - inclusion bodies
What are two ways to detect the presence of a virus via cultivation?
Cultivation/isolation in cell culture or by egg inoculation
What kind of microscope is necessary to detect viruses that can not be grown in-vitro?
Electron microscope
Can use negative staining
What are the differences between the transmission and scanning microscope?
Transmission: Method based on transmitted elections: 2d image, seeks to see what is inside or beyond the surface, has higher magnification and greater resolution
Scanning: Method is based on scattered electrons. Focuses on the sample’s surface and its composition. **3D image
T/F: A transmission electron microscope yields a 3-D image
FALSE
Scanning electron microscope = 3D
Transmission = 2D
What is a gold standard test?
A dx test that is considered to be the most accurate and best available under a particular condition or set of conditions
Sensitivity is based on..?
The probability that cases WITH the infection will have a POSITIVE result using the test under evaluation
What is specificity based on..?
The probability that cases WITHOUT the infection will have a NEGATIVE result under test evaluation
How do you collect/prepare a serum sample?
Use a red top tube/vacutainer, allow the sample to clot, centrifuge the sample, supernatant = serum
How do you collect/prepare a plasma sample?
Use a lavender top EDTA tube/vacutainer, centrifuge, supernatant = plasma
Vacutainer tubes used to collect blood samples to obtain serum for diagnostic purposes usually have ___ colord caps?
Red
Blue
Lavender
Green
RED
What are the 6 steps of the basic Enzyme-linked immunosorbent assay (ELISA) test?
- antigen coated well
- Add antibody tagged with an enzyme
- Antigen binds to enzyme-tagged antibody
- Wash the excess unbound antiodies
- Add substrate
- Enzyme tagged to antibody which is bound to antigen will chnage color of substrate. Intensity of color indicates more positive reaction
How does the direct ELISA work?
Antigens are immobilized and enzyme-conjugated primary antibodies are used to detect or quantify antigen concentration. The specificity of the primary antigen is very important
How does the indirect ELISA work?
Primary antibodies are not labeled, but detected instead with enzyme-conjugate secondary antibodies that recognize the primary antibodies.
How does the sandwich ELISA work?
The antigen to be measures is bound between a layer of capture antibodies and a layer of detection antibodies. The two antibodies must be very critically chosen to prevent cross-ractivity or competition of binding sites
What ELISA has a more positive result, when you see a weaker color change?
Competitive ELISA
A decrease signal when compared to assay wells with purified antigen alone, indicates the presence of antigens in the sample
T/F: When using a competitive ELISA, a weaker signal indicates the presence of antigens in a sample
TRUE
How does direct Fluorescence antibody test (FAT) work?
Labelled antibodies are added onto the sample (antigen). Visible fluorescence appears at the binding site of the specific antibodies
How does indirect FAT work?
IFAT employs a secondary antibody labeled with a fluorescent marker that recognizes the primary antibody bound to antigen
How does immunohistochemistry work?
The antibody is tagged with an enzyme, generally horseradish peroxidase. The enzyme reacts with a substrate to produce a colored product that can be visualized in the infected cells with a standard light microscope
T/F: Immunochromatograhy is a point of care test
TRUE
A POC (point of care) test is one that is simple to perform, easy to carry, and does not require specialized equipment