Objective To optimize experimental conditions of the enrichment and concentration of viruses in purified water with mixed cellulose ester (MCE) membrane adsorption-ultrafiltration method.
Methods Bacteriophage MS2 was used as the target virus. First, water sample cation concentration, MCE membrane pore size, type of eluent, and elution method were included in the orthogonal experiment on the MCE adsorption of bacteriophage MS2. Then, the optimized conditions were further screened by applying centrifugal force of 3 000 and 4 000 grams during the ultrafiltration process. Reverse transcription-fluorescence quantitative polymerase chain reaction (RT-qPCR) was used in the detection and the recovery rates of bacteriophage MS2 under various conditions were calculated and analyzed statistically.
Results The optimal enrichment conditions for MCE-ultrafiltration method were as follows: water sample cation concentration of 0.025 mol/L, MCE membrane pore size of 0.22 μm), using 1% Tris-glycine-beef extract (TGBE) solution in elution, adopting vibration method in elution, use of ultrafiltration tubes retaining organics with the molecular weight of 30 kD in the ultrafiltration, and centrifugal force of 3 000 grams. A recovery rate of 81.20% (95% confidence interval: 71.86% – 90.54%) for bacteriophage MS2 viruses in purified water samples was achieved under the above conditions in the experiment.
Conclusion The optimized membrane adsorption MCE-ultrafiltration method could enrich bacteriophage MS2 viruses in purified water samples at a high recovery rate and be used in the enrichment and concentration of viruses in purified water.
DownLoad: