The E-Test (AB Biodisk, Solna, Sweden), a new and novel approach developed to test antimicrobial susceptibility, was compared with the agar dilution method. A collection of 57 organisms, including those with known resistances to various antimicrobial agents was tested against 15 drugs using the National Committee for Clinical Laboratory Standard (NCCLS) methods. The E-Test quantitative accuracy (+/-2 log2 dilutions) compared with agar dilution results was 99% for Neisseria gonorrhoeae, 95% for Escherichia coli strains with extended-spectrum beta-lactamases, 100% for Enterobacter-Citrobacter spp. with type-1 stably derepressed enzymes, and 95% for Enterococcus spp. with various drug resistances (penicillins, ciprofloxacin, gentamicin, and vancomycin). The E-Test procedures with these species were easy to perform, reproducible, and quantitatively and qualitatively accurate (89%-100%). The E-Test appears to be an acceptable option for susceptibility testing of these difficult-to-treat, drug-resistant bacterial strains.