A highly efficient second-step concentration technique for bacteriophages and enteric viruses using ammonium sulfate and Tween 80.
Addition of Tween 80 to a 1.5% solution of beef extract was found to enhance the elution of bacteriophages adsorbed to electronegative filters. When reconcentration of the eluate was attempted by ammonium sulfate precipitation, a floating layer containing most of the viruses was formed. This floating layer can be obtained with several nonionic detergents including Tween 80 and under a salt saturation of 55% with ammonium sulfate, potassium tartrate, and sodium phosphate. Virus recovery ranged from 91 to 103% and was obtained with several bacteriophage strains. With poliovirus type 1, coxsackievirus B-4, and rotavirus SA-11 the recoveries were 100, 20, and 80%, respectively, but toxicity to cell culture was encountered: after removal of the detergent by a second floating layer method the recovery was 32% for poliovirus. Compared with organic flocculation, this method also had both improved recovery for bacteriophages and protective properties for samples frozen at -70 degrees C.
Armon
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Centre de recherche en virologie, Institut Armand Frappier, Laval, Québec, Canada.
Arella
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Payment
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D000327:Adsorption; D000645:Ammonium Sulfate; D000818:Animals; D001435:Bacteriophages / Q000302:isolation & purification*; D011232:Chemical Precipitation; D029822:Enterovirus B, Human / Q000302:isolation & purification*; D017955:Poliovirus / Q000302:isolation & purification*; D011136:Polysorbates; D012401:Rotavirus / Q000302:isolation & purification*; D014871:Water Microbiology
D011136:Polysorbates; D000645:Ammonium Sulfate