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DIFFERENTIAL EFFECT OF TETRAZOLIUM UPON BACTERIOPHAGE PLAQUE ASSAY TITERS
Citation:
Hurst, C., J. Blannon, AND R. Hardaway. DIFFERENTIAL EFFECT OF TETRAZOLIUM UPON BACTERIOPHAGE PLAQUE ASSAY TITERS. U.S. Environmental Protection Agency, Washington, D.C., EPA/600/J-94/428 (NTIS PB95122628), 1994.
Description:
This study examined whether the practice of incorporating either tetrazolium red ortetrazolium violet dye into plaque assay medium deleteriously influences plaque assay titers. epresentative members of six different virus families were studied: ystoviridae (06), Leviviridae (MS2), Microviridae (OX174), Myoviridae (T2), Podoviridae (P22), and Siphoviridae (Denver, T1, VD13). or the purpose of this study we initially used 300 uG/mL as the final dye concentration. his represents a moderately high concentration as determined by examining the literature for bacteriophage assays. ffect of the dye was determined based upon titer ratio values (titer with dye in medium divided by titer without dye in medium). he criteria established for determining suppression of any individual virus type were that a reduction in viral titer of more than 5 percent (ratio <0.95) had to occur in 4 of 5 independent replicate trials for that virus type, and also that both the mean and median of the, five ratio values representing the trials for that virus type had to be below 0.85. ased upon these criteria, each of the members of the Podoviridae and Siphoviridae families appeared to be suppressed by either one or both dyes at the 300 uG/mL concentration. he chosen representatives of the other bacteriophage families were not suppressed by either dye at the 300 uG/mL concentration. ubsequent trials revealed no suppression of Podoviridae or Siphoviridae plaque assay titers when tested with these same two dyes at the lower concentrations of 150 uG/mL and 50 uG/mL. nterestingly, the bacteriophage families whose members were affected by the dyes have additional commonality in that these are the two bacteriophage families whose members possess both double stranded DNA genomes and non-contractile tails.
