Bloodstream infection (BSI) is a systemic infectious disease that can lead to shock, disseminated intravascular coagulation, multiorgan failure, and even death. Blood culture is considered the gold standard for the etiological diagnosis of BSI
however, blood culture is time-consuming and has a low positivity rate, which has limited its utility for early and rapid clinical diagnosis. Nanopore sequencing technology (NST), a third-generation sequencing method, offers rapid detection, real-time single-molecule sequencing, and ultra-long reads. These features enable the prompt detection of pathogens and the analysis of drug-resistant genes and genomic characteristics, thereby optimizing the clinical diagnosis and treatment of BSI. In this article, we summarize the application of NST in the etiological diagnosis of BSI.