Isolation, Cultivation, and Identification of Viruses

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Isolation, Cultivation, and Identification of Viruses

Viral Identification

Polymerase Chain Reaction (PCR): SARS and West Nile Virus IDed

restriction fragment length polymorphisms (RFLPs)

Serological method = Western blotting commonly used

IDed by reaction with antibodies

need electron microscope

Growing Animal Viruses in the Lab

Lines

a lot of samples are sent to central labs to avoid above problem

major problem is that cell lines must be kept free of microbial contamination

Continuous Cell Lines

HeLa cell line cancer from woman who died in 1951

immortal cell lines

indefinite number of generations

Primary Cell Lines

diploid cell lines from human enbryos can be used for 100 generations

die out after a few generations

derived from tissue slices

In Cell Structures

cytopathic effect (CPE): cells of monolayer deteriorate as they multiply

more convenient to work with

preferred type of growth medium

In Embryonated Eggs

why asked if allergic to eggs before receiving vaccination

viral growth

pocks and lesions on egg membrane

embryo cell damage

death of embryo

In Living Animals

recent studies with mice have been used to study replication

some human viruses can't be grown in animals: AIDS

inoculation for diagnosis

Growing Bacteriophages in the Lab

can be grown on solid media

plaque forming units (PFU)

plaque method: clearings and turbidity

can be grown in liquid suspension