Improved detection and quantification of mouse cytomegalovirus by real-time PCR.
During latent cytomegalovirus (CMV) infection, viral presence cannot be detected by plaque assay. Therefore, we assessed the applicability of real-time PCR for temporal determination of virus dissemination in two different mouse strains. Eight-week-old BALB/c and C57BL/6J mice were infected with mouse CMV (MCMV) and sacrificed at 1, 2, 4, 6, 14 and 28 days post infection. Real-time PCR was used to determine MCMV copy number in the heart, bone marrow cells, aorta and blood. In lung, liver, salivary gland and spleen the presence of MCMV was determined both by plaque assay and real-time PCR. In analogy with the plaque assay, the real-time PCR technique revealed higher numbers of MCMV genomic copies in all organs obtained from BALB/c mice when compared with C57BL/6J mice, demonstrating the applicability of the technique. A significant correlation was observed between both assays when a positive test result was seen with both assays. Nonetheless, lower viral infectivity titers were found compared to real-time PCR data. Thus, the real-time PCR technique is more sensitive in detecting the presence of MCMV and is therefore well suited for (dose-response) intervention studies aimed at studying virus eradication.
Vliegen
Inge
I
Department of Medical Microbiology, University Hospital Maastricht, P.O. Box 5800, 6202 AZ Maastricht, The Netherlands. ivl@lmib.azm.nl
Herngreen
Selma
S
Grauls
Gert
G
Bruggeman
Cathrien
C
Stassen
Frank
F
D000818:Animals; D004279:DNA, Viral / Q000032:analysis; D006566:Herpesviridae Infections / Q000821:virology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018146:Muromegalovirus / Q000235:genetics / Q000302:isolation & purification*; D009928:Organ Specificity; D016133:Polymerase Chain Reaction / Q000379:methods*; D010948:Viral Plaque Assay
D004279:DNA, Viral