ObjectiveTo establish a rapid screening method for influenza virus neuraminidase（NA） inhibitors sourced from Chinese medicines based on fluorescence detection. MethodThe method was constructed based on the principle that after the reaction of the test sample and a certain amount of NA， the activity of some NA will be inhibited by the test sample， and the NA that is still active after the addition of the substrate can generate fluorescence at a specific wavelength when combined with the fluorescent substrate， and the inhibition rate of the test sample on NA was calculated according to the measured fluorescence intensity， so as to evaluate the in vitro inhibitory activity of the test sample on NA. A total of 49 high-purity chemical components from 12 Chinese medicines were used to evaluate the in vitro anti-NA activity by the established method. The theoretical calculated values of binding energy and inhibition constant after docking between the NA protein receptor and the test sample were used to prove the accuracy of the experimental results. The established method was applied to detect the in vitro NA inhibitory activity of different batches of Banlangen granules and Kangbingdu granules， so as to evaluate the quality consistency among different batches of samples. ResultThe methodological examination results showed that the method had good accuracy and repeatability. The screening results of 49 components showed that 22 of them had strong in vitro inhibitory activity against NA than peramivir ［half inhibitory concentration（IC₅₀） was 131.2 μmol·L^-1］， such as schaftoside， isoorientin， chebulinic acid， menthone and isoschaftoside. The inhibitory activity of the remaining 27 components was weaker than that of peramivir. The molecular docking results showed that the theoretical calculation results of binding energies and inhibition constants of most compounds were basically consistent with the experimental results. The test results of the inhibitory activity of 12 batches of Banlangen granules on NA showed that the quality consistency among samples A1， A2， B2， C1， C2， E2 and F2 was good. The analysis results of the inhibitory activity of 9 batches of Kangbingdu granules produced by the same manufacturer on NA showed that the inhibitory rates of samples K1 to K9 were 37.68%， 36.18%， 31.37%， 33.98%， 40.36%， 33.76%， 40.69%， 41.08%， 40.06% when the concentration of 0.02 g·mL^-1， and the average inhibitory rate was 37.24%. ConclusionIn this paper， we successfully established an analytical method that can be used to rapidly evaluate whether Chinese medicines （derived from chemical components of traditional Chinese medicine or proprietary Chinese medicines） have in vitro anti-NA activity， which can be a powerful supplement to the existing screening methods for influenza virus NA inhibitors. And this method was used to screen 22 compounds from 12 Chinese medicines with good in vitro inhibitory activity against NA， which can provide candidate compounds for the development of anti-influenza small molecule drugs.