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Chapter 98 : Human Cytomegalovirus

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Human Cytomegalovirus, Page 1 of 2

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Abstract:

The human cytomegalovirus (CMV), formally designated human herpesvirus 5 (HHV-5) by the International Committee on Taxonomy of Viruses, is a member of the family Herpesviridae, which includes herpes simplex virus types 1 (HHV-1) and 2 (HHV-2), varicella-zoster virus (HHV-3), Epstein-Barr virus (HHV-4), and human herpesviruses 6, 7, and 8. Purified blood leukocytes are used when performing the CMV antigenemia assay, while whole blood, plasma obtained from anticoagulated whole blood, serum obtained from clotted blood, or purified peripheral blood leukocytes have all been used to quantify CMV DNA in molecular amplification assays. The antigenemia assay is relatively simple to perform and is based on immunocytochemical detection of the 65-kDa lower-matrix phosphoprotein (pp65) in the nuclei of peripheral blood leukocytes. PCR is currently the most widely used molecular method for the detection of CMV DNA and mRNAs, and the sensitivity and specificity of PCR for diagnosis of active CMV infection have been evaluated. The spin-amplification shell vial assay is based on the amplification of virus in cell cultures after low-speed centrifugation and detects viral antigens produced early in the replication of CMV, before the development of cytopathic effect (CPE). Confirmation of herpes B virus in culture can be done using monoclonal antibodies or molecular techniques. PCR for herpes B virus is generally preferred over culture for diagnostic purposes, since it is comparatively rapid, is highly sensitive and specific, and avoids the need to amplify infectious virus to high titers.

Citation: Hodinka R. 2011. Human Cytomegalovirus, p 1558-1574. 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.ch98

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Figures

Image of FIGURE 1
FIGURE 1

Fixed hematoxylin-eosin-stained lung tissue from a patient with interstitial pneumonia. Note the numerous giant cells that possess large intranuclear inclusions surrounded by characteristically clear halos (arrows). Less pronounced granular inclusions may also be present in the cytoplasm (inset). (Courtesy of Eduardo Ruchelli.)

Citation: Hodinka R. 2011. Human Cytomegalovirus, p 1558-1574. 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.ch98
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Image of FIGURE 2
FIGURE 2

CMV antigen-positive polymorphonuclear leukocytes. Note the nuclear staining when a monoclonal antibody directed against pp65 is used. (A) Magnification, x100; (B) magnification, x400.

Citation: Hodinka R. 2011. Human Cytomegalovirus, p 1558-1574. 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.ch98
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Image of FIGURE 3
FIGURE 3

CPE produced by a CMV isolate in human skin fibroblasts 10 days postinoculation. Unstained preparation; magnification, x100. (Courtesy of Sergio Stagno.)

Citation: Hodinka R. 2011. Human Cytomegalovirus, p 1558-1574. 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.ch98
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Image of FIGURE 4
FIGURE 4

Demonstration of CMV early antigens in the nuclei (arrows) of infected MRC-5 cells following shell vial culture and IFA staining. (A) Staining of immediate-early antigen appears as an even matte green fluorescence with specks of brighter green. (B) Viral inclusions (owl's eyes) may be visible in the nuclei. Magnification, x400.

Citation: Hodinka R. 2011. Human Cytomegalovirus, p 1558-1574. 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.ch98
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Tables

Generic image for table
TABLE 1

Available commercial CMV antigenemia assays

CMV Brite, CMV Brite Turbo, and CINAkit are FDA licensed for qualitative testing only; Light Diagnostics kit is not FDA licensed

Citation: Hodinka R. 2011. Human Cytomegalovirus, p 1558-1574. 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.ch98
Generic image for table
TABLE 2

Available commercial molecular assays for CMV

Abbreviations: pol, polymerase; pp65, phosphoprotein 65; MIE, major immediate early; IE1, immediate early 1; ASR, analyte-specific reagent; RUO, research use only.

Citation: Hodinka R. 2011. Human Cytomegalovirus, p 1558-1574. 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.ch98

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