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Chapter 10.7 : Direct Detection of Viruses and in Clinical Samples

Editor: Amy L. Leber1
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Affiliations: 1: Editor in chief, Department of Laboratory Medicine, Nationwide Children’s Hospital, Columbus, Ohio
Content Type: Reference
Publication Year: 2016

Category: Clinical Microbiology

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Abstract:

The direct detection of viruses and in specimens can be performed with a variety of methods, including rapid antigen detection assays and immunofluorescence methods ( Table 10.7–1 ), as well as electron microscopy, cytopathology, and histopathology. Numerous molecular assays have also been developed and many are commercially available for diagnostic purposes; these are extensively described in other sections of this book. Rapid antigen detection methods for viral diagnostics are almost universally performed with commercially available kits in accordance with manufacturer’s instructions, most commonly for influenza viruses, respiratory syncytial virus (RSV), and rotavirus. This procedure therefore focuses on IF methods with brief sections on electron microscopy and cytohistopathology, and is revised from the previously published edition ( ).

Citation: Leber A. 2016. Direct Detection of Viruses and in Clinical Samples, p 10.7.1-10.7.10. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch10.7
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Direct Detection of Viruses and Chlamydia in Clinical Samples
Citation: Leber A. 2016. Direct Detection of Viruses and in Clinical Samples, p 10.7.1-10.7.10. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch10.7
/content/10.1128/9781555818814.chap10.7.fig10.7-1
Figure 10.7–1
Preparation of aspirates and fluids for IF.
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Image of Figure 10.7–1
Figure 10.7–1

Preparation of aspirates and fluids for IF.

Citation: Leber A. 2016. Direct Detection of Viruses and in Clinical Samples, p 10.7.1-10.7.10. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch10.7
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Figure 10.7–2
Preparation of swabs, scrapings, and impression smears for IF.
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Image of Figure 10.7–2
Figure 10.7–2

Preparation of swabs, scrapings, and impression smears for IF.

Citation: Leber A. 2016. Direct Detection of Viruses and in Clinical Samples, p 10.7.1-10.7.10. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch10.7
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Figure 10.7–3
Leukocyte preparation with several cells showing intense CMV pp65 staining and a few cells (arrows) showing weak speckled nuclear staining (×400). Provided by Light Diagnostics.
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Image of Figure 10.7–3
Figure 10.7–3

Leukocyte preparation with several cells showing intense CMV pp65 staining and a few cells (arrows) showing weak speckled nuclear staining (×400). Provided by Light Diagnostics.

Citation: Leber A. 2016. Direct Detection of Viruses and in Clinical Samples, p 10.7.1-10.7.10. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch10.7
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Figure 10.7–4
(A) Kidney tubule with characteristic “owl eye” inclusion of CMV (arrow). Courtesy of W. L. Drew. (B) Giemsa stain of conjunctival cells showing cytoplasmic inclusion (arrow) of Courtesy of W. L. Drew. (C) Cytologic examination (Tzanck smear) of scrapings from the base of an ulcerative HSV lesion showing multinucleated giant cells. Courtesy of W. L. Drew. (D) Large eosinophilic inclusions fill the cytoplasms of several infected basal cells (molluscum bodies). Courtesy of W. L. Drew.
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10.1128/9781555818814/fig10.7-4.gif
Image of Figure 10.7–4
Figure 10.7–4

(A) Kidney tubule with characteristic “owl eye” inclusion of CMV (arrow). Courtesy of W. L. Drew. (B) Giemsa stain of conjunctival cells showing cytoplasmic inclusion (arrow) of Courtesy of W. L. Drew. (C) Cytologic examination (Tzanck smear) of scrapings from the base of an ulcerative HSV lesion showing multinucleated giant cells. Courtesy of W. L. Drew. (D) Large eosinophilic inclusions fill the cytoplasms of several infected basal cells (molluscum bodies). Courtesy of W. L. Drew.

Citation: Leber A. 2016. Direct Detection of Viruses and in Clinical Samples, p 10.7.1-10.7.10. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch10.7
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References

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1. Clarke L. 2010. Direct detection of viruses and Chlamydia in clinical samples. In Garcia LS (ed), Clinical Microbiology Procedures Handbook, 3rd ed. ASM Press, Washington, DC.
2. Payne WJ Jr, Marshall DL, Shockley RK, Martin WJ. 1988. Clinical laboratory applications of monoclonal antibodies. Clin Microbiol Rev 1: 313– 329.
3. Matthey S, Nicholson D, Ruhs S, Aiden B, Knock M, Schultz K, Schmuecker A. 1992. Rapid detection of respiratory viruses by shell vial culture and direct staining by using pooled and individual monoclonal antibodies. J Clin Microbiol 30: 540– 544.
4. Stout C, Murphy MD, Lawrence S, Julian S. 1989. Evaluation of a monoclonal antibody pool for rapid diagnosis of respiratory viral infections. J Clin Microbiol 27: 448– 452.
5. Landry ML, Ferguson D. 2000. SimulFluor respiratory screen for rapid detection of multiple respiratory viruses in clinical specimens by immunofluorescence staining. J Clin Microbiol 38: 708– 711.
6. CDC. 1991. False-positive results with the use of chlamydia tests in the evaluation of suspected sexual abuse. Morb Mortal Wkly Rep 39: 932– 935.
7. Goudswaard F, Sabbe L, van Belzen C. 1989. Interference by gram-negative bacteria in the enzyme immunoassay for detecting Chlamydia trachomatis. J Infect Dis 18: 94– 96.
8. Hammerschlag MR, Rettig PJ, Shields ME. 1988. False positive results with the use of chlamydia antigen detection tests in the evaluation of suspected sexual abuse in children. Pediatr Infect Dis J. 7: 11– 14
9. Poder K, Sanchez N, Robin PM, McHugh M, Hammerschlag MR. 1989. Lack of specificity of Chlamydiazyme for detection of vaginal chlamydial infection in prepubertal girls. Pediatr Infect Dis J 8: 358– 360.
10. Pratt BC, Tait IA, Anyaegbunam WL. 1989. Rectal carriage of Chlamydia trachomatis in women. J Clin Pathol 42: 1309– 1310.
11. Biel SS, Gelderblom HR. 1999. Diagnostic electron microscopy is still a timely and rewarding method. J Clin Virol 13: 105– 119.
12. Doane FW, Anderson N. 1987. Electron Microscopy in Diagnostic Virology: Practical Guide and Atlas. Cambridge University Press, New York, NY.
13. Fong CKY. 1994. Electron microscopy and immunoelectron microscopy, p 99– 107. In Hsiung GD, Fong CKY, Landry ML (ed), Hsiung’s Diagnostic Virology, 4th ed. Yale University Press, New Haven, Conn.
14. Miller SE. 1986. Detection and identification of viruses by electron microscopy. J Electron Microsc Technol 4: 265– 301.
15. Lamson DM, Ramani R, Kleabonas M, Metcalfe M, Humphrey C, St George K. 2014. An unusual case of influenza-like illness after yellow fever vaccination. J Clin Virol 60: 67– 69.
16. Hsiung GD. 1994. Histocytochemical staining, p 83– 90. In Hsiung GD, Fong CKY, Landry ML (ed), Hsiung’s Diagnostic Virology, 4th ed. Yale University Press, New Haven, CN.
17. Kusne S, Mañez R, Frye BL, St George K, Abu-Elmagd K, Tabasco-Menguillon J, Fung JJ, Todo S, Rinaldo C, Ehrlich GD. 1997. Use of DNA amplification for diagnosis of cytomegalovirus enteritis after intestinal transplantation. Gastroenterology 112: 1121– 1128.

Tables

Direct Detection of Viruses and Chlamydia in Clinical Samples
Citation: Leber A. 2016. Direct Detection of Viruses and in Clinical Samples, p 10.7.1-10.7.10. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch10.7
/content/10.1128/9781555818814.chap10.7.table10.7-1
Table 10.7–1
Direct antigen detection for viral and chlamydial infections
Generic image for table
Table 10.7–1

Direct antigen detection for viral and chlamydial infections

Citation: Leber A. 2016. Direct Detection of Viruses and in Clinical Samples, p 10.7.1-10.7.10. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch10.7
/content/10.1128/9781555818814.chap10.7.table10.7-2
Table 10.7–2
Troubleshooting IF problems
Generic image for table
Table 10.7–2

Troubleshooting IF problems

Citation: Leber A. 2016. Direct Detection of Viruses and in Clinical Samples, p 10.7.1-10.7.10. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch10.7
/content/10.1128/9781555818814.chap10.7.table10.7-3
Table 10.7–3
Inclusion morphology
Generic image for table
Table 10.7–3

Inclusion morphology

Citation: Leber A. 2016. Direct Detection of Viruses and in Clinical Samples, p 10.7.1-10.7.10. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch10.7

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