Specific assays for bacteria using phage mediated release of adenylate kinase.
A sensitive and rapid assay method for the specific detection of bacteria was developed using Escherichia coli and Salmonella newport as the test organisms. Bacteriophages were used to provide specific lysis of the bacteria and then the release of cell contents was measured by ATP bioluminescence. Increased sensitivity was obtained by focusing on the bacteria's adenylate kinase (AK) as the cell marker instead of ATP as conventionally used. Fewer than 10(3) E. coli cells could be readily detected in less than 1 h. Salmonella newport assays, although as sensitive, were slower and took up to 2 h. The effects of the culture medium, the phage, and the presence of non-specific bacteria were examined.
Blasco
R
R
MAFF Central Science Laboratory, Sand Hutton, York, UK.
Murphy
M J
MJ
Sanders
M F
MF
Squirrell
D J
DJ
D000263:Adenylate Kinase / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D001431:Bacteriological Techniques; D001433:Bacteriolysis; D001435:Bacteriophages; D004926:Escherichia coli / Q000201:enzymology* / Q000302:isolation & purification / Q000821:virology; D008163:Luminescent Measurements; D012475:Salmonella / Q000201:enzymology* / Q000302:isolation & purification / Q000821:virology; D012680:Sensitivity and Specificity; D013997:Time Factors
D001426:Bacterial Proteins; D000263:Adenylate Kinase