Laboratory Diagnosis of Cytomegalovirus
المؤلف:
Stefan Riedel, Jeffery A. Hobden, Steve Miller, Stephen A. Morse, Timothy A. Mietzner, Barbara Detrick, Thomas G. Mitchell, Judy A. Sakanari, Peter Hotez, Rojelio Mejia
المصدر:
Jawetz, Melnick, & Adelberg’s Medical Microbiology
الجزء والصفحة:
28e , p492
2025-11-16
45
A. Culture, Polymerase Chain Reaction, and Antigen Detection Assays
PCR assays are used to detect circulating virus in blood, CSF, and urine. PCR assays can provide quantitative viral load data, which can be used to predict CMV disease in immunocompromised patients. Monoclonal antibodies against viral antigens can be used to detect virus-positive leukocytes from patients.
B. Isolation of Virus
Human fibroblasts are used for virus isolation attempts. The virus can be recovered most readily from tissues, body fluids, throat washings, and urine. In cultures, 2–3 weeks are usually needed for the appearance of cytologic changes, consisting of small foci of swollen, translucent cells with large intranuclear inclusions (see Figure 1C and D). Shell vial cultures allow for detection of CMV antigen using fluorescent antibodies prior to development of cytopathic effect for a more rapid diagnosis.
Fig1. Cytopathic effects induced by herpesviruses. A: Herpes simplex virus in HEp-2 cells (hematoxylin and eosin stain, 57×), with early focus of swollen, rounded cells. B: Varicella-zoster virus in human kidney cells (hematoxylin and eosin stain, 228×), with multinucleated giant cell containing acidophilic intranuclear inclusions (arrow). C: Cytomegalovirus in human fibroblasts (unstained, 35×) with two foci of slowly developing cytopathic effect. D: Cytomegalovirus in human fibroblasts (hematoxylin and eosin stain, 228×), showing giant cells with acidophilic inclusions in the nuclei (small arrow) and cytoplasm (large arrow), the latter being characteristically large and round. (Courtesy of I Jack; reproduced from White DO, Fenner FJ: Medical Virology, 3rd ed. Academic Press, 1986.)
C. Serology
Many types of assays can detect CMV IgG antibodies, indicative of past infection (and the potential to undergo reactivation). Detection of viral IgM antibodies suggests a recent infection. Serologic assays are not informative for immunocompromised patients. Furthermore, serologic techniques cannot distinguish strain differences among clinical isolates.
D. Resistance Testing
CMV resistance testing involves sequencing of the viral kinase (UL97) and DNA polymerase (UL54) genes and com paring to a database of known resistance mutations. UL97 mutations can confer resistance to ganciclovir, while UL54 mutations can confer resistance to ganciclovir, cidofovir, and foscarnet.
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