We developed a novel immunochromatographic assay (ICA) (EZ-Step VanA rapid kit; Dinona, Korea) for the detection of VanA ligase from vancomycin-resistant enterococci (VRE). Of eight monoclonal antibodies screened by ELISAs, the VanA ligase ICA constructed with 1H9 plus 3G11 showed the greatest reactivity. The detection limit of the kit was $6.3{ imes}10^6$ CFU per test. Of 127 vancomycin-resistant microorganisms, 100 vanA VRE were positive in the VanA ligase ICA, and 27 non-vanA vancomycin-resistant isolates were negative. These results were consistent with those of the PCR analyses. Thus, our ICA is a reliable and easy-to-use immunological assay for detecting VanA-producing VRE in clinical laboratories.