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Chapter 80 : Reagents, Stains, Media, and Cell Cultures: Virology

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Reagents, Stains, Media, and Cell Cultures: Virology, Page 1 of 2

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Abstract:

Despite the integration of nucleic acid-based diagnostic methods into routine clinical virology laboratory practices, direct fluorescent-antibody assays (DFAs) using monoclonal antibodies and culture-based methods are still an integral part of viral diagnostics. DFAs can provide a rapid diagnosis when used directly with clinical samples. In addition, DFAs are used for confirmation of virus identity when cytopathic effect is noted in traditional cell culture and to identify viral proteins by blind staining when rapid centrifugation-enhanced cell culture is used. Cell culture methods are useful for the detection of novel or unsuspected viral agents, for documentation of active infection, to obtain isolates for antiviral susceptibility testing, to assess response to antiviral therapy, for serologic strain typing, and for vaccine and therapeutic clinical trials. This chapter reviews the reagents, stains, tissue culture media, and traditional cell culture methods currently used in diagnostic virology laboratories. Rapid cell culture methods using single cell lines, cocultured cell lines, and transgenic cell lines are described. Quality control and safety regarding the handling and culturing of virology samples is discussed.

Citation: Ginocchio C, Van Horn G, Harris P. 2015. Reagents, Stains, Media, and Cell Cultures: Virology, p 1422-1431. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch80
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Figures

Image of FIGURE 1
FIGURE 1

Immunofluorescence detection of in McCoy cells. Magnification, ×200. Courtesy of Quidel/Diagnostic Hybrids. doi:10.1128/9781555817381.ch80.f1

Citation: Ginocchio C, Van Horn G, Harris P. 2015. Reagents, Stains, Media, and Cell Cultures: Virology, p 1422-1431. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch80
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Image of FIGURE 2
FIGURE 2

Immunofluorescence detection of respiratory pathogens in R-Mix cells using D3 Ultra Kit reagents (Quidel/Diagnostic Hybrids). (A) Uninoculated R-Mix cells; (B) adenovirus; (C) influenza A virus; (D) influenza B virus; (E) parainfluenza virus type 1; (F) parainfluenza virus type 2; (G): parainfluenza virus type 3; (H) RSV. Magnification, ×170. Courtesy of Quidel/Diagnostic Hybrids. doi:10.1128/9781555817381.ch80.f2

Citation: Ginocchio C, Van Horn G, Harris P. 2015. Reagents, Stains, Media, and Cell Cultures: Virology, p 1422-1431. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch80
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Image of FIGURE 3
FIGURE 3

Detection of HSV-1 and HSV-2 in ELVIS cells (Quidel/Diagnostic Hybrids). (A) Blue cells positive for HSV with X-Gal stain; (B) immunofluorescence of uninoculated ELVIS cells; (C) HSV-1-positive ELVIS immunofluorescence (note nuclear pattern); (D) HSV-2-positive ELVIS immunofluorescence (note cytoplasmic pattern). Magnification, ×170. Courtesy of Quidel/Diagnostic Hybrids. doi:10.1128/9781555817381.ch80.f3

Citation: Ginocchio C, Van Horn G, Harris P. 2015. Reagents, Stains, Media, and Cell Cultures: Virology, p 1422-1431. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch80
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Image of FIGURE 4
FIGURE 4

Immunofluorescence detection of respiratory pathogens from clinical specimens using Duet stains (Quidel/Diagnostic Hybrids). (A and B) Influenza A Duet stain (influenza A [gold] and respiratory pool [green] for adenovirus; influenza B; parainfluenza virus types 1, 2, and 3; and RSV): negative cells (A); influenza A-positive cells (B). (C) RSV Duet stain (RSV [gold] and respiratory pool [green] for adenovirus; influenza A; influenza B; and parainfluenza viruses 1, 2, and 3): RSV-positive and influenza A-positive cells. Magnification, ×170. Courtesy of Quidel/Diagnostic Hybrids. doi:10.1128/9781555817381.ch80.f4

Citation: Ginocchio C, Van Horn G, Harris P. 2015. Reagents, Stains, Media, and Cell Cultures: Virology, p 1422-1431. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch80
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Image of FIGURE 5
FIGURE 5

Detection of coxsackie B virus in Super E-Mix cells (Quidel/Diagnostic Hybrids). (A) Unstained, uninoculated cells; (B) unstained coxsackie B virus CPE; (C and D) immunofluorescence staining with pan-enterovirus antibody pool of uninoculated Super E-Mix cells (C) and coxsackie B virus-infected cells (D). Magnification, ×170. Courtesy of Quidel/Diagnostic Hybrids. doi:10.1128/9781555817381.ch80.f5

Citation: Ginocchio C, Van Horn G, Harris P. 2015. Reagents, Stains, Media, and Cell Cultures: Virology, p 1422-1431. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch80
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Image of FIGURE 6
FIGURE 6

Immunofluorescence detection of family viruses in H&V-Mix cells (Quidel/Diagnostic Hybrids). (A) Uninoculated H&V-Mix cells; (B) CMV; (C) VZV; (D) HSV-1; (E) HSV-2. Magnification, ×170. Courtesy of Quidel/Diagnostic Hybrids. doi:10.1128/9781555817381.ch80.f6

Citation: Ginocchio C, Van Horn G, Harris P. 2015. Reagents, Stains, Media, and Cell Cultures: Virology, p 1422-1431. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch80
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References

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Tables

Generic image for table
TABLE 1

Commercially available DFA and IFA reagents for the detection of chlamydiae and viruses

Citation: Ginocchio C, Van Horn G, Harris P. 2015. Reagents, Stains, Media, and Cell Cultures: Virology, p 1422-1431. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch80
Generic image for table
TABLE 2

List of cell lines and virus susceptibility profiles

Citation: Ginocchio C, Van Horn G, Harris P. 2015. Reagents, Stains, Media, and Cell Cultures: Virology, p 1422-1431. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch80
Generic image for table
TABLE 3

List of cocultured cell lines and virus susceptibility profiles

Citation: Ginocchio C, Van Horn G, Harris P. 2015. Reagents, Stains, Media, and Cell Cultures: Virology, p 1422-1431. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch80

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