Objective:To develop a sandwich ELISA for quantitative detection enolase of Candida albicans,and apply the assay to detect enolase levels in supernatant of common fungi cultures. To investigate the potential diagnostic value of enolase in invasive candidiasis. Methods:Anti-enolase of Candida albicans monoclonal antibody was employed as coating antibody,HRP-conjugated goat polyclonal antibody against Candida albicans enolase was used as detecting antibody. The performance parameters of the sandwich ELISA including precision,linear range and limit of detection were verified by using recombinant Candida albicans enolase. Then the developed assay was applied to determine enolase levels in supernatant of pathogenic fungi cultures such as common Candida spp. Cryptococcus neoformans and Saccharomyces cerevisiae for preliminary evaluation. Results:The intra and inter-coefficient of variation was 6.61%,9.19% and 6.98%,13.81% at the concentration of 20 ng/ml and 5 ng/ml respectively. The limit of detection was 1.25 ng/ml. The linear range was 1.25~50.00 ng/ml. The level of enolase in Candida albicans culture after incubated in 37℃ for 24 h was 3.06 ng/ml and gradually increased to 33.43 ng/ml at 120 h after incubated in 37℃,consistent with growth of hyphae. The sandwich ELISA was weak cross-reactive to Candida parapsilosis and no cross-reactivity to Candida tropicalis,Candida guilliermondii,Candida glabrata,Cryptococcus neoformans and Saccharomyces cerevisiae was found. Conclusion:A sandwich ELISA for quantitative detection enolase of Candida albicans was developed,which had the potential to be applied to stndy invasive candidasis.