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Laboratory Diagnosis of Infectious Diseases Principles and Practice pp 39–59Cite as

Virus Isolation and Identification

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Abstract

Cell Cultures: Monolayer cultures of primary, diploid, and continuous cell lines are the hosts of choice for virus isolation. Quality cell cultures are available commercially and are conveniently maintained in the laboratory. After proper decontamination and purification, each clinical sample is inoculated into several types of cell cultures; the preferred lines vary from virus to virus.

Alternative Cell Culture Techniques: New cell culture techniques have been developed to enhance virus isolation. Shell vials containing a monolayer of cells growing on a cover slip provide a virus isolation system that can be placed in the centrifuge and spun to enhance virus infection. Following incubation, the infected cover slip is withdrawn and stained to confirm virus identification. Shell vials are helpful in cytomegalovirus isolation. Herpes simplex isolation and identification systems are commercially available. A culture of herpes-susceptible cells and an immunostaining system are included. Inclusion of microspheres in cell culture has been shown to provide increased surface area for attachment of cells. Increased cell numbers provide for increased virus production. Several viruses can be isolated in suspensions of human lymphocytes rather than in traditional monolayer cultures.

Virus Quantitation: Viruses are quantitated by titration and plaquing methods. Formulas by Reed-Muench (1938) and Karber (1931) are used to determine the end point from titration data.

Virus Detection and Identification: Both immunologic and nonimmunologic methods are used in identification of isolated viruses and for detection of viral antigen within cell cultures or within clinical materials.

Nonimmunologic methods include histologic staining, electron microscopy, hemadsorption (HAD), hemagglutination (HA), challenge interference (CI), and DNA probes. HAD, HA, and CI are used primarily to detect non-cytopathogenic effect-producing viruses within cell cultures. DNA probes provide the most specific viral identification and will probably replace many traditional techniques.

Immunologic methods include immunofluorescence (IF), immunoperoxidase (IP), enzyme-linked immunosorbent assay (ELISA), neutralization (NEUT), hemagglutination inhibitation (HAI), passive agglutination (PA), radioimmunoassay, and immune adherence hemagglutination. IF, IP, ELISA, and PA provide rapid and accurate virus identification, whereas NEUT and HAI, although accurate, are more costly and timeconsuming.

Keywords

  • Herpes Simplex
  • Respiratory Syncytial Virus
  • Virus Isolation
  • Cover Slip
  • Hemorrhagic Fever With Renal Syndrome

These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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  1. Diane S. Leland
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Leland, D.S., French, M.L.V. (1988). Virus Isolation and Identification. In: Laboratory Diagnosis of Infectious Diseases Principles and Practice. Springer, New York, NY. https://doi.org/10.1007/978-1-4612-3900-0_3

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  • DOI: https://doi.org/10.1007/978-1-4612-3900-0_3

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