Isolation, Cultivation, and Identification of Viruses
Growing Bacteriophages in the Lab
can be grown in liquid suspension
can be grown on solid media
plaque method: clearings and turbidity
plaque forming units (PFU)
Growing Animal Viruses in the Lab
In Living Animals
inoculation for diagnosis
some human viruses can't be grown in animals: AIDS
recent studies with mice have been used to study replication
In Embryonated Eggs
viral growth
death of embryo
embryo cell damage
pocks and lesions on egg membrane
why asked if allergic to eggs before receiving vaccination
In Cell Structures
preferred type of growth medium
more convenient to work with
cytopathic effect (CPE): cells of monolayer deteriorate as they multiply
Lines
Primary Cell Lines
derived from tissue slices
die out after a few generations
diploid cell lines from human enbryos can be used for 100 generations
Continuous Cell Lines
indefinite number of generations
immortal cell lines
HeLa cell line cancer from woman who died in 1951
major problem is that cell lines must be kept free of microbial contamination
a lot of samples are sent to central labs to avoid above problem
Viral Identification
need electron microscope
Serological method = Western blotting commonly used
IDed by reaction with antibodies
restriction fragment length polymorphisms (RFLPs)
Polymerase Chain Reaction (PCR): SARS and West Nile Virus IDed