INTRODUCTION: Leishmaniasis, a protozoan disease caused by infection by METHODS: With the development of metagenomic nextgeneration sequencing (mNGS) technology for pathogen identification, it is possible to detect specific nucleic acid sequences characteristic of RESULTS: Our report demonstrates that the mNGS method is applicable to peripheral blood samples (PB), which are far more readily available in clinical settings, in addition to bone marrow aspirate samples (BM), which are traditionally used for diagnosis of visceral leishmaniasis. CONCLUSION: Our report validates the efficacy of mNGS technology as a rapid and accurate method of diagnosis for leishmaniasis.