L9 Lab diagnosis Flashcards
What is the direct approach to viral diagnosis
Demonstration of virus in material from patient approach
What is the indirect approach to viral diagnosis
Demonstration of an immune response to virus (serology)
How are viruses isolated
In tissue culture as need to grow in cells
How is tissue culture grown
In flask lying flat down
Cells stick to plastic until confluence growth and scrap cells off surface
Red liquid - cells
What do uninflected human embryonic lung fibroblasts look like under light microscope
Long thin shape of cells
What do rhinovirus infected lung fibroblasts look like
Round and shrivelled typical of rhinovirus
Cytopathic effect CPE
What do cytomegalovirus infected fibroblasts look like
Causes rounded and swollen cells for CPE effect
What is important when trying to isolate many viruses
Not all cells have a receptive for every virus - need to provide a range of different cell types if trying to isolate many viruses
What are Hep2 cells infected with respiratory syncytial virus like
Huge gaps in cell sheet
Large multi nucleated giant cells
Virus caused these cells to fuse together - syncytia
How is a genital ulceration tested
Scrap cells off swab
Break swab into transport medium as need cells alive not dried
What is in transport medium
Antibiotics to stop overgrowth
Where are viral antigens expressed
In cells as viruses replicate in cells
What is early antigen fluorescent foci
Antigen expressed early in cell cycle
Make antibody for this and tag with fluorescent marker
What is virus isolation like
Can be slow
Why is detection of early antigen fluorescent foci good
Can do within 24 hrs rather than waiting for CPEs which take long
What are the pros of virus isolation in tissue culture
Positive result unequivocal
What are the cons of virus isolation in tissue culture
May be slow
Can’t be universal (can be hard to grow in cell culture)
How is virus isolation sped up
Haemaosdrption
Detection of early antigen fluorescent foci