Abstract: Objective To evaluate the value of valve metagenomie next-generation sequencing ( mNGS) in the pathogenie diagnosis of infective endocarditis (IE). Methods A total of 25 patients who underwent valve surgery in our hospital between June 1, 2022 and November 30, 2022 were enrolled in the study. According to the modifed Duke criteria, 20 patients were diagnosed as IE patients and 5 patients were non-IE patients. The venous blood and valve tissue samples fom each patient were collected, and blood culure, valve culture and mNGS were performed. The elinical eficacy of mNGS and traditional diagnostic methods such as blood culture and valve culture in diagnosing the etiology of IE was compared. Results Among the 20 patients diagnosed with E by pathology, 8 were positive for blood culture, 3 were positive for valve culture, and 19 were positive for valve mNGS. The sensitivity of blood culture, valve culture and valve mNGS in the diagnosis of IE were 40%，15% and 95%，respectively. The detection results of valve mNGS and blood culture or valve culture were consistent at the genus level of bacteria. The relative abundance of genera ( P=0.001), number of species sequences ( P= 0.028), and coverage rate ( P= 0.003) of valve mNGS were related to the duration of antibiotic use. Valve mNGS could guide the antibiotic therapy in IE patients with negative culture. Conclusion Valve mNGS has high clinical value in the pathogenie diagnosis of IE, which may provide a basis for subsequent antibiotic treatment.