Summary
Diethylaminoethyl-dextran (DEAE-D) greatly increases the detection of infectious RNA in MK cells. The DEAE-D is the basis of a simple method of plaque assay for poliovirus RNA under isotonic conditions that is 100 times more sensitive than conventional methods. The same polycationic substance yields a 3 to 4-fold increase in plaque counts when used in the assay of intact poliovirus.
Temperature and duration of adsorption have little influence on the DEAE-D method of assay of RNA. A somewhat lesser, but definite, enhancing effect is obtained when the assay cells are exposed to DEAE-D and washed repeatedly before inoculation with RNA.
The presence of DEAE-D in extractions of poliovirus RNA with cold phenol and sodium dodecyl sulfate may improve the yield of infectious RNA. A total of 105.6 PFU of Mahoney RNA were detected in the extract made directly from 4 ml. of infected MK cells with a titer of 108.3 PFU/ml. The absolute titer in the ethanol precipitate was 107 PFU of RNA per ml. determined with the DEAE-D method.
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Dedicated to the Honor of the 60th birthday of ProfessorSven Gard.
This work was supported in part by USPHS Research Grant AI 01799 from the Institute of Allergy and Infectious Diseases and the Milton J. Greenman Fund.
Research Associate, Department of Virus Research, Karolinska Institutet, 1961–62.
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Pagano, J.S., Vaheri, A. Enhancement of infectivity of poliovirus RNA with diethylaminoethyl-dextran (DEAE-D). Archiv f Virusforschung 17, 456–464 (1965). https://doi.org/10.1007/BF01241201
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DOI: https://doi.org/10.1007/BF01241201