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A highly efficient second-step concentration technique for bacteriophages and enteric viruses using ammonium sulfate
1Centre de recherche en virologie, Institut Armand Frappier, Laval, Québec, Canada.
Canadian Journal of Microbiology
|May 1, 1988
Summary
This study introduces a novel method using nonionic detergents like Tween 80 to efficiently elute and recover bacteriophages from filters. The technique enhances virus recovery and offers protective properties for frozen samples.
Area of Science:
- Virology
- Biochemistry
- Microbiology
Background:
- Bacteriophages are viruses that infect bacteria.
- Efficient recovery of viruses from samples is crucial for research and diagnostics.
- Electronegative filters are commonly used for virus adsorption.
Purpose of the Study:
- To develop an improved method for eluting and concentrating bacteriophages from electronegative filters.
- To evaluate the efficacy of nonionic detergents and salt precipitation for virus recovery.
- To compare this new method with existing techniques like organic flocculation.
Main Methods:
- Addition of Tween 80 to beef extract to enhance bacteriophage elution from filters.
- Concentration of eluates using ammonium sulfate precipitation to form a virus-containing floating layer.
- Testing various nonionic detergents and salt saturation levels (ammonium sulfate, potassium tartrate, sodium phosphate).
- Assessing virus recovery rates for different bacteriophage strains and human viruses (poliovirus, coxsackievirus, rotavirus).
- Evaluating the protective properties of the method for frozen samples.
Main Results:
- Tween 80 significantly enhanced bacteriophage elution.
- A floating layer formed upon ammonium sulfate precipitation, containing the majority of viruses.
- High virus recovery rates (91-103%) were achieved for bacteriophages.
- Variable recoveries were observed for human viruses (poliovirus 100%, coxsackievirus B-4 20%, rotavirus SA-11 80%).
- Detergent removal was necessary to mitigate cell culture toxicity, reducing poliovirus recovery to 32%.
- The method demonstrated improved bacteriophage recovery and protective properties compared to organic flocculation.
Conclusions:
- Nonionic detergents, particularly Tween 80, combined with salt precipitation, offer an effective method for bacteriophage elution and concentration.
- This technique provides superior recovery and sample protection for bacteriophages compared to traditional methods.
- Further optimization is needed for efficient recovery of certain human viruses due to potential toxicity issues.