title: Development of a highly sensitive quantitative competitive PCR assay for the detection of murine cytomegalovirus DNA.
abstract: Viral persistence and molecular latency are characteristic of infection by the human cytomegalovirus (HCMV). Using the murine cytomegalovirus (MCMV) as a model for human infection, a quantitative-competitive polymerase chain reaction (QC-PCR) assay was developed to detect and quantify MCMV-DNA in the salivary glands of infected mice. The QC-PCR detected high numbers of MCMV DNA copies in the absence of infectious virus. By comparing the DNA content and the results obtained from a standard semiquantitative plaque assay, it is concluded that 1 plaque-forming unit (pfu) is the equivalent of approximately 1500 viral genomes. By day 42-post infection (pi) 4x10(3) copies of DNA/1 mg tissue were sufficient to reactivate infectious virions after cyclophosphamide immunosupression. By day 90 pi, however, when the DNA load was decreased to <1.2x10(2), reactivation was not observed. These results indicate that viral reactivation will occur when the number of infectious DNA copies is equivalent about 2-3 pfu. This quantitative test may therefore help to detect CMV and the risk of reactivation in immunosupressed patients.
authors.affiliation: Department of Oral Biology, Faculty of Dental Medicine, Hebrew University, Jerusalem, Israel. apalmon@cc.huji.ac.il
authors.forename: A
authors.initials: A
authors.lastname: Palmon
authors.forename: S
authors.initials: S
authors.lastname: Tel-or
authors.forename: E
authors.initials: E
authors.lastname: Shai
authors.forename: B
authors.initials: B
authors.lastname: Rager-Zisman
authors.forename: Y
authors.initials: Y
authors.lastname: Burstein
mesh_terms: D000818:Animals; D003520:Cyclophosphamide / Q000494:pharmacology; D004279:DNA, Viral / Q000032:analysis*; D004195:Disease Models, Animal; D005260:Female; D006566:Herpesviridae Infections / Q000276:immunology / Q000821:virology*; D006801:Humans; D016867:Immunocompromised Host; D007166:Immunosuppressive Agents / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D018146:Muromegalovirus / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012469:Salivary Glands / Q000821:virology; D012680:Sensitivity and Specificity; D014775:Virus Activation; D017735:Virus Latency
chemical_list: D004279:DNA, Viral; D007166:Immunosuppressive Agents; D003520:Cyclophosphamide