Interleukin 6 (IL-6) is an interleukin that acts as both a proinflammatory and anti-inflammatory cytokine. It can be used as a potential diagnostic biomarker for sepsis. The aim of this study was to establish an easy-to-use detection kit for rapid, quantitative and on-site detection of IL-6. To develop the new IL-6 quantitative detecting kit, a double-antibody sandwich immunofluorescent assay was employed based on europium nanoparticles (Eu-np) combined with lateral flow immunoassay (LFIA). The performance of the new developed kit was evaluated in the aspects of parallel analysis, linearity, sensitivity, precision, accuracy, specificity and clinical sample analysis. Two-hundred and fourteen serum samples were used to carry out the clinical sample analysis. The new IL-6 quantitative detecting kit exhibited a wide linear range (2-500 pg/mL) and a good sensitivity (0.37 pg/mL). The intra-assay coefficient of variation (CV) and the inter-assay CV were 5.92%-8.87% and 7.59%-9.04%, respectively. The recovery rates ranged from 102% to 106%. Furthermore, a high correlation (n = 214, r = 0.9756, p < 0.01) was obtained when compared with SIEMENS CLIA IL-6 kit. Thus, the new quantitative method for detecting IL-6 has been successfully established. The results indicated that the newly-developed strip based on Eu-np combined with LFIA was a facile, fast, highly sensitive, low-cost, reliable biosensor and suitable for rapid and point-of-care test (POCT) for IL-6 in serum. Copyright © 2019 Elsevier Inc. All rights reserved.