A simple micro-growth assay for enumerating bacteria

J Microbiol Methods. 2003 Apr;53(1):77-86. doi: 10.1016/s0167-7012(02)00226-9.

Abstract

A simple method for nonspecific determination of bacteria concentrations in a variety of liquid samples was developed. The assay was based on the time required for a sample grown in liquid media to reach a threshold turbidity. Samples were combined with media in a covered 96-well microwell plate and the turbidity was monitored in real time as the bacteria grew in a temperature-controlled plate reader. A significant problem with growth in microwells was condensation on the cover, which prevented accurate turbidity measurement. This problem was overcome by coating the cover with a small amount of surface-active agent. Salmonella and E. coli concentrations could be determined with a relative error of approximately 20% at levels from 10 to 10(6) cells/ml (eight replicates). An assay of 10 samples with standards required 10 min to set up and 20 min for data processing using a computer spreadsheet program. Growth time at 37 degrees C ranged from 4 h for samples at 10(7) cells/ml to 16 h for samples at 10 cells/ml.

MeSH terms

  • Algorithms
  • Bacteria / growth & development*
  • Bacteriological Techniques / methods
  • Binding Sites, Antibody
  • Calibration
  • Cell Division
  • Colony Count, Microbial / methods*
  • Salmonella enteritidis / growth & development
  • Salmonella enteritidis / immunology
  • Salmonella enteritidis / isolation & purification