Objective To explore the clinical value of metagenomic next-generation sequencing (mNGS) in the diagnosis and treatment of severe and complex infection of malignant hematological disorder. Methods The mNGS test results, traditional etiology test results and general clinical data of inpatients with malignant hematological disorder in the Department of Hematology, the Affiliated Hospital of Southwest Medical University between June 2020 and February 2022 were retrospectively analyzed. To explore the clinical application value of mNGS in the diagnosis and treatment of severe complicated infection of hematological disorder. Results A total of 21 patients were included. The samples included 18 peripheral blood samples, 2 pleural fluid samples and 1 alveolar lavage fluid sample. In the included patients, through mNGS, pathogenic bacteria were directly detected in 17 patients, including 8 fungi, 9 bacteria and 10 viruses, of which 9 were mixed infections. The positive rate (81.0% vs. 33.3%, P=0.002), sensitivity (85.7% vs. 30.0%), granulocytopenia (9 vs. 3 cases, P=0.031) and the types of pathogen (Z=−3.416, P=0.001) detected by mNGS were all higher than those by traditional method. The infection control of 17 patients improved in varying degrees after adjusting the treatment plan according to the test results. ConclusionsmNGS has significantly higher detection rate and sensitivity for bacteria, fungi, viruses and mixed infections. Compared with the traditional method, mNGS has more efficient characteristics. Its clinical application can further improve the diagnosis and treatment efficiency of severe complicated infection of malignant hematological disorder, and thus improve the survival rate of patients.