A. Palmon et al., Development of a highly sensitive quantitative competitive PCR assay for the detection of murine cytomegalovirus DNA, J VIROL MET, 86(2), 2000, pp. 107-114
Viral persistence and molecular latency are characteristic of infection by
the human cytomegalovirus (HCMV). Using the murine cytomegalovirus (MCMV) a
s a model for human infection, a quantitative-competitive polymerase chain
reaction (QC-PCR) assay was developed to detect and quantify MCMV-DNA in th
e salivary glands of infected mice. The QC-PCR detected high numbers of MCM
V DNA copies in the absence of infectious virus. By comparing the DNA conte
nt and the results obtained from a standard semiquantitative plaque assay,
it is concluded that 1 plaque-forming unit (pfu) is the equivalent of appro
ximately 1500 viral genomes. By day 42-post infection (pi) 4 x 10(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 < 1.2 x 10(2), reactivation was not observed. These resul
ts indicate that viral reactivation will occur when the number of infectiou
s DNA copies is equivalent about 2-3 pfu. This quantitative test may theref
ore help to detect CMV and the risk of reactivation in immunosupressed pati
ents. (C) 2000 Elsevier Science B.V. All rights reserved.