Detection of invasiveness of mammalian cells by Escherichia coli: collaborative study.

In a collaborative study of 2 standard procedures (Procedure I, presented in J. Assoc. Off. Anal. Chem. 60, 546-562 (1977) and Procedure II in the Bacteriological Analytical Manual, 1978), 7 laboratories evaluated the invasive potential of Escherichia coli. Monolayers of HeLa cultures were infected with E. coli (infectivity ratio 100 bacteria/mammalian cell) suspended in 2 menstrua: heat-inactivated fetal bovine serum (Procedure I) and 0.2% bovine albumin Fraction V dissolved in Earle's buffered salts (Procedure II). After uptake of the bacteria, intracellular growth, and differential staining, the percentages of HeLa cells containing a minimum of 5 bacteria were determined microscopically. All laboratories correctly identified invasiveness of E. coli by Procedure I; 5 of 6 laboratories recognized invasiveness by Procedure II. Although Procedure I was more reproducible than Procedure II, repeatability did not differ significantly. Procedure I was adopted official first action for presumptive recognition of invasiveness in E. coli.