Improved detection of bacteria by flow cytometry using a combination of antibody and viability markers.
A proprietary fluorogenic marker for cell viability (Chemchrome) was investigated for the detection of bacteria using flow cytometry. This marker was used in combination with fluorescently labelled monoclonal antibodies (against Salmonella typhimurium). Owing to the former's broad band emission spectrum, it was necessary to use the novel dye RED613 for the antibodies. This combined protocol, being sensitive only to the live Salm. typhimurium cells, reduced errors due to intrinsic fluorescence and non-specific binding. Detection of the order of 100 cells ml-1 was achieved in 30 min. This level was achieved even in the presence of large numbers of non-target or dead organisms.
Clarke
R G
RG
Department of Food Biophysics, Institute of Food Research, Norwich Laboratory, UK.
Pinder
A C
AC
D000911:Antibodies, Monoclonal; D015169:Colony Count, Microbial; D004396:Coloring Agents; D004926:Escherichia coli / Q000254:growth & development / Q000276:immunology / Q000302:isolation & purification*; D005434:Flow Cytometry / Q000379:methods*; D005516:Food Microbiology; D008856:Microscopy, Fluorescence; D011933:Reagent Kits, Diagnostic; D012486:Salmonella typhimurium / Q000254:growth & development / Q000276:immunology / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods*
D000911:Antibodies, Monoclonal; D004396:Coloring Agents; D011933:Reagent Kits, Diagnostic