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Chapter 77 : Reagents, Stains, Media, and Cell Cultures: Virology

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Abstract:

Some molecular assays, for practical reporting purposes, may take as long as overnight cell culture and may not lend themselves to single-specimen testing as well as cell culture does. For these reasons, cell cultures are still an indispensible research and clinical laboratory tool, particularly when combined with the use of highly specific monoclonal antibodies (MAbs) for the detection of common viruses and spp. or when cell lines are engineered to produce virus-induced enzymes, such as beta-galactosidase for the detection of herpes simplex viruses (HSV). This chapter describes the cell lines, reagents, stains, and media used in association with traditional tube and rapid viral culture techniques. Definitive identification of certain viruses or Chlamydia spp. can be directly determined in clinical samples when using MAbs labeled with fluorescein isothiocyanate, methylrhodamine isothiocyanate, or phycoerythrin with an Evans blue and/or propidium iodide counterstain. The lot number and date of use for all media, buffers, reagents, and additives should also be recorded. An important consideration in using cell culture is ensuring collection of cellular material and maintaining the viability of the organisms from the time of sample collection to inoculation in cell culture. Cell culture media are an important part of the production and maintenance of cells.

Citation: Ginocchio C, Harris P. 2011. Reagents, Stains, Media, and Cell Cultures: Virology, p 1289-1296. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch77

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References

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Tables

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TABLE 1

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

Refer to manufacturers' websites for FDA and in vitro diagnostic status of reagents. Although uses for the reagents may be suggested on manufacturers' websites, all suggested applications may not have been validated by the manufacturer. Abbreviations: adeno, adenovirus; CC, culture confirmation; DSD, direct specimen detection; dual, a 2-fluorophore stain; FluA, influenza virus A; ID, identification; IF, immunofluorescence; Para, parainfluenza virus; IFA, indirect fluorescent antibody; HSV-1/2, HSV type 1 or 2.

Respiratory panel A: adenovirus, influenza viruses A and B, parainfluenza viruses 1, 2, and 3, RSV, HMPV; respiratory panel B: adenovirus, influenza viruses A and B, parainfluenza viruses 1, 2, and 3, RSV; respiratory panel C: adenovirus, influenza viruses A and B, parainfluenza viruses 1, 2, and 3, and differential identification of RSV; respiratory panel D: adenovirus, influenza virus B, parainfluenza viruses 1, 2, and 3, RSV, and differential detection of influenza virus A.

Citation: Ginocchio C, Harris P. 2011. Reagents, Stains, Media, and Cell Cultures: Virology, p 1289-1296. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch77
Generic image for table
TABLE 2

List of cell lines and virus susceptibility profiles

Primary cell cultures.

Available as fresh and frozen ReadyCells (Quidel/Diagnostic Hybrids, Inc.).

Abbreviations: HCoV, human coronavirus; EBV, Epstein-Barr virus; SARS-CoV, severe acute respiratory syndrome coronavirus.

Citation: Ginocchio C, Harris P. 2011. Reagents, Stains, Media, and Cell Cultures: Virology, p 1289-1296. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch77
Generic image for table
TABLE 3

List of cocultured cell lines and virus susceptibility profiles

Abbreviations: SARS-CoV, severe acute respiratory syndrome coronavirus.

Available from Quidel/Diagnostic Hybrids, Inc.

Available as both fresh and frozen cells which do not require propagation but are ready to use.

Citation: Ginocchio C, Harris P. 2011. Reagents, Stains, Media, and Cell Cultures: Virology, p 1289-1296. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch77

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