Bacteriophage titration of E. coli in vivo panel with agar overlay assay (AOA)

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DOCUMENTS/REFERENCES
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BIOLOGICAL INFORMATION
BACKGROUND INFORMATION
The agar overlay assay (AOA) for phage titration is used to determine the concentration of a phage product measured as plaque-forming units per mL against a panel of E. coli strains (ATCC 25922, ATCC 700336, AR Bank #0349, and AR Bank #0055).
NUMBER OF ASSAYS ON PANEL
4
ASSAYS ON PANEL
FAMILY NAME ITEM NUMBER PROTEIN NAME ORGANISM ASSAY SUB TYPE
Bacteriophage titration of E. coli (ATCC 25922) with agar overlay assay (AOA)697011 Gram-negative Bacteria
Bacteriophage titration of E. coli (ATCC 700336) with agar overlay assay (AOA)697012 Gram-negative Bacteria
Bacteriophage titration of E. coli (AR Bank #0349) with agar overlay assay (AOA)697013 Gram-negative Bacteria
Bacteriophage titration of E. coli (AR Bank #0055) with agar overlay assay (AOA)697014 Gram-negative Bacteria
ORGANISM
Gram-negative Bacteria
ASSAY INFORMATION
ASSAY TYPE
Functional
TESTING INFORMATION
PROCEDURE SUMMARY
The testing phage suspension is 10-fold serially diluted with phage dilution buffer (such as SM buffer) for a total of 6 concentrations (typical range is 10⁴ to 10⁹ dilutions). A 0.1 mL aliquot of each phage dilution is mixed with 0.1 mL of bacterial overnight culture in a broth medium. Duplicate phage-bacteria mixtures are prepared with one phage dilution series. Phage absorption to cells is allowed at room temperature for a period of 15 minutes. Molten top agar medium, containing 0.75% agar and with a volume of 3 mL, is added to the phage-bacteria mixture, and the tube is mixed by quick inversion. The contents are plated onto pre-warmed (25°C) bottom agar medium, typically LB containing 1.5% agar. The plates are rotated to evenly distribute the top agar and then allowed to solidify at room temperature. The plates are then incubated for 18 – 24 h at 35-37°C. Plaques are counted on plates with 1 to 300 distinguishable plaques to calculate PFU/mL values.
CRITERIA FOR SIGNIFICANCE
The agar overlay assay (AOA) for phage titration is used to determine the concentration of a phage product measured as plaque forming units per mL (PFU/mL).
TURNAROUND TIME
STANDARD
20 Days
CLINICAL RELEVANCE
THERAPEUTIC AREA
Infectious Disease
ADDITIONAL INFORMATION
BRAND
PDS
TESTING LOCATION
Taiwan - Taipei

 

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