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Chapter 69 : Enhanced Detection of Viruses in Cell Cultures

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Abstract:

Several viruses such as herpes simplex virus (HSV), respiratory syncytial virus (RSV), influenza virus, and cytomegalovirus (CMV) were able to infect cells cultured in vitro, but the time for recognition of the cytopathic effects (CPE) produced by these agents ranged from a few days to several weeks. In addition to the age of cells, condition of cell monolayers, and specificity of monoclonal antibodies for immediate-early or early viral antigens, other important variables include incubation temperature, number of shell vial cell cultures used per specimen, centrifugation, type of specimen (urine, blood, bronchoalveolar lavage (BAL) fluid, tissue), quality of fluorescence equipment, chemical pretreatment of cell mono-layers, and technical experience with the assay to subjectively evaluate specific results. Several comparisons have demonstrated that the shell vial assay is as sensitive as (for HSV and respiratory tract viruses [adenoviruses, parainfluenza virus types 1, 2, and 3, enterovirus, influenza A and B virus, measles virus, mumps virus, and RSV]) or even more sensitive than (for CMV) the recovery of these viruses in conventional tube cell cultures, which may require several days to weeks for recognition by CPE. The diagnostic laboratory may always find a use for cell cultures; however, the next level of test performance in the clinical laboratory will be formatted for the automated extraction and quantitation detection of target nucleic acids. Amplified nucleic acids will be monitored in real time by thermocycling instruments designed to be used in routine biosafety level 2 laboratories.

Citation: Smith T. 2006. Enhanced Detection of Viruses in Cell Cultures, p 617-625. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch69

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Human immunodeficiency virus 1
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Nucleic Acid Amplification Techniques
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Severe Acute Respiratory Syndrome
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FIGURE 1

Detection of early antigens in the shell vial assay by immunofluorescence (left) and immunoperoxidase (right) stains. Magnification, ×400.

Citation: Smith T. 2006. Enhanced Detection of Viruses in Cell Cultures, p 617-625. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch69
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References

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Tables

Generic image for table
TABLE 1

Technical variables in the shell vial assay

Citation: Smith T. 2006. Enhanced Detection of Viruses in Cell Cultures, p 617-625. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch69
Generic image for table
TABLE 2

Sensitivity of detection of CMV DNA by real-time PCR compared to antigenemia results

Citation: Smith T. 2006. Enhanced Detection of Viruses in Cell Cultures, p 617-625. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch69
Generic image for table
TABLE 3

Laboratory diagnosis of HSV and VZV infections by cell cultures and by real-time PCR

Citation: Smith T. 2006. Enhanced Detection of Viruses in Cell Cultures, p 617-625. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch69

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