The algorithm for a new identification system was designed on the basis of colony color and morphology on CHROMagar Orientation medium in conjunction with simple biochemical tests such as indole (IND), lysine decarboxylase (LDC), and ornithine decarboxylase (ODC) utilization tests with gram-negative bacilli isolated from urine samples as well as pus, stool, and other clinical specimens by the following colony characteristics, biochemical reactions, and serological results: pinkish to red, IND positive (IND⁺), Escherichia coli; metallic blue, IND⁺, LDC⁺, and ODC negative (ODC), Klebsiella oxytoca; IND⁺, LDC, and ODC⁺, Citrobacter diversus; IND⁺ or IND, LDC, and ODC, Citrobacter freundii; IND, LDC⁺, and ODC⁺, Enterobacter aerogenes; IND, LDC, and ODC⁺, Enterobacter cloacae; IND, LDC⁺, and ODC, Klebsiella pneumoniae; diffuse brown and IND⁺, Morganella morganii; IND, Proteus mirabilis; aqua blue, Serratia marcescens; bluish green and IND⁺, Proteus vulgaris; transparent yellow-green, serology positive, Pseudomonas aeruginosa; clear and serology positive, Salmonella sp.; other colors and reactions, the organism was identified by the full identification methods. The accuracy and cost-effectiveness of this new system were prospectively evaluated. During an 8-month period, a total of 345 specimens yielded one or more gram-negative bacilli. A total of 472 gram-negative bacillus isolates were detected on CHROMagar Orientation medium. For 466 of the isolates (98.7%), no discrepancies in the results were obtained on the basis of the identification algorithm. The cost of identification of gram-negative bacilli during this period was reduced by about 70%. The results of this trial for the differentiation of the most commonly encountered gram-negative pathogens in clinical specimens with the new algorithm were favourable in that it permitted reliable detection and presumptive identification. In addition, this rapid identification system not only significantly reduced costs but it also improved the daily work flow within the clinical microbiology laboratory.