Using a novel qEBV assay, we demonstrated that young adults with primary EBV infection rapidly cleared virus from blood but not from the oropharynx. High oral concentrations of EBV in asymptomatic persons who have resumed normal activities support the concept that infectious mononucleosis is most likely acquired by kissing.
Quantification of Epstein-Barr virus (EBV) in peripheral blood is important for the diagnosis and management of serious EBV diseases, including posttransplant lymphoproliferative disorder. A variety of PCRbased methods are currently in use; however, there is little information on their comparability. This study assessed the relative performance of different quantitative assays. A multicenter comparative study was performed at eight sites using three panels consisting of serial dilutions of quantified EBV DNA and extracts from a total of 19 whole-blood specimens. Samples were distributed and tested blindly. Instrumentation, probe chemistries, amplification targets, and other test-related aspects varied considerably between laboratories. Each laboratory's calibration curve indicated strong evidence of a consistent log-linear relationship between viral load and cycle threshold, suggesting that intralaboratory tracking of a given patient would yield similar relative quantitative trends among the participating test sites. There was strong concordance among laboratories with respect to qualitative test results; however, marked quantitative discordance was seen. For most samples, the across-laboratory interquartile range of the reported viral load (in copies/l) was roughly 0.6 log-units, and for one sample the overall range was approximately 4.2 log-units. While intralaboratory tracking of patients may yield similar results, these data indicate a need for caution when attempting to compare clinical results obtained at different institutions and suggest the potential value to be gained by more standardized testing methodology.
Glial cells of the retina are anatomically distinctive and are thought to contribute importantly to retinal electrophysiology. However, no adequate preparation exists for studying them in isolation, in vitro. This report provides guidelines for primary retinal glial cultures (RET) and compares basal tissue culture features with those for neocortical glia (CX) and the well-studied rat glial line, C6. Cell attachment and growth of RET, CX, and C6 are unique. These differences are explored by the use of specific media and sera. RET attachment, unlike that for CX or C6, was far more sensitive to medium than serum. RET cells attached least quickly, CX most quickly; 4 hr after plating 20% of RET remained unattached. RET growth was poor and relatively insensitive to medium. In contrast, growth of CX or C6 was medium dependent. Serum had substantial effects on the growth of all three glial lines. Pig, goat, horse, and dog sera were particularly effective, often comparing favorably to fetal calf serum. Medium or serum optimal for cell attachment, typically, was not optimal for growth and serum effects were more dramatic than those of medium. By all measures, CX and C6, both derived from brain, were more alike than were the two rabbit primaries, CX and RET. The data reveal substantial differences between presumably similar cells and indicate a need for an empirically based choice of both basal-salt media and serum to optimize specific aspects of cell development in culture.
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