The use of DNA probes in studies of human cytomegalovirus.

Abstract

Hybridization assays provide a sensitive and rapid means for studying the molecular biology of viral replication and for identifying viral nucleic acid in biological specimens. Such assays are attractive because the detection of virus does not require intact virions or concomitant viral protein synthesis, both of which may be absent in a latently infected cell or in a virus-associated tumor. For molecular and clinical studies on human cytomegalovirus (HCMV), we have cloned and characterized subgenomic EcoRI fragments representative of the entire genome of HCMV strain AD169. To study the epidemiology of HCMV infections and to identify the presence of HCMV nucleic acid in urine, blood, Kaposi's sarcoma, and other tissues, we have used various hybridization techniques, including DNA dot/slot-blot hybridization, Southern blot hybridization, and in situ cytohybridization. These studies demonstrate how cloned molecular probes can be used to study the molecular biology, pathogenesis, and treatment of viral infections.

Cytomegalovirus infection induces high levels of cyclins, phosphorylated Rb, and p53, leading to cell cycle arrest

“…VOL. 69,1995 CELL CYCLE ARREST CAUSED BY HCMV 6701 (77). It is therefore tempting to speculate that the delay in cyclin A synthesis observed in our system is responsible for the induction of MPF at early times and the premature entry into mitosis.…”

“…Human cytomegalovirus (HCMV), a member of the herpesvirus family, is the leading viral cause of birth defects and poses a serious health threat to immunocompromised persons (for reviews, see references 4, 33, 51, 60, and 67). In AIDS, HCMV is a common opportunistic infection (12,44,45,68,69) and is implicated as a possible cofactor in the progression of the disease. Although HCMV infection is usually asymptomatic in the healthy individual, it is suspected that activation of latent virus may trigger cellular damage in infected organs of certain susceptible individuals.…”

Diagnostic deoxyribonucleic acid probes for infectious diseases.

“…Probes to a wide spectrum of DNA-and RNA-containing viruses have been developed and used successfully to extend our knowledge of the natural history of these agents as well as for diagnostic tools. Probes to the herpes viruses (2,3,13,16,20,54,74,88,103,115,122,125,126,143), adenoviruses (71,90,142), enteroviruses (58,108,109), reoviruses (26), hepatitis viruses (6,8,9,67,114), papillomaviruses (5, 96), papovaviruses (44), and parvoviruses (22) have all been described.…”

Viral infections in the acquired immunodeficiency syndrome

“…In situ hybridization with CMV nucleic acid probes has also proven to be a useful adjunct in the diagnosis of CMV infection in tissue and urine specimens. This technque is capable of detecting latently infected cells (Chou and Merigan, 1983;Myerson et al, 1984;Spector and Spector, 1985). It has been used to demonstrate CMV in colonic tissue that was negative for the virus on routine histologic and immunofluorescence examination (Roche et al, 1981).…”