biostudies-literatureStahl-Rommel SNational Aeronautics and Space AdministrationNHGRI NIH HHS106https://www.ebi.ac.uk/biostudies/studies/S-EPMC7830261biostudies-literatureUnknown12(1)For the past two decades, microbial monitoring of the International Space Station (ISS) has relied on culture-dependent methods that require return to Earth for analysis. This has a number of limitations, with the most significant being bias towards the detection of culturable organisms and the inherent delay between sample collection and ground-based analysis. In recent years, portable and easy-to-use molecular-based tools, such as Oxford Nanopore Technologies' MinION™ sequencer and miniPCR bio's miniPCR™ thermal cycler, have been validated onboard the ISS. Here, we report on the development, validation, and implementation of a swab-to-sequencer method that provides a culture-independent solution to real-time microbial profiling onboard the ISS. Method development focused on analysis of sGenesReal-Time Culture-Independent Microbial Profiling Onboard the International Space Station Using Nanopore Sequencing.PMC7830261ISS Vehicle Office; Office of the Chief Technologist; Advanced Exploration System Life Support SystemsR01 HG010485Turner DJPaten BStahl-Rommel SJain MNguyen HNAunon-Chancellor SMArnold RRBurton ASStoddart DAkeson MSharp GMJohn KKCastro-Wallace SLJuul SCastro CLfalseReal-Time Culture-Independent Microbial Profiling Onboard the International Space Station Using Nanopore Sequencing.For the past two decades, microbial monitoring of the International Space Station (ISS) has relied on culture-dependent methods that require return to Earth for analysis. This has a number of limitations, with the most significant being bias towards the detection of culturable organisms and the inherent delay between sample collection and ground-based analysis. In recent years, portable and easy-to-use molecular-based tools, such as Oxford Nanopore Technologies' MinION™ sequencer and miniPCR bio's miniPCR™ thermal cycler, have been validated onboard the ISS. Here, we report on the development, validation, and implementation of a swab-to-sequencer method that provides a culture-independent solution to real-time microbial profiling onboard the ISS. Method development focused on analysis of s2021-01-01T00:00:00Z2021 Jan2026-04-29T21:28:41.631Z2025-04-04T10:55:59.817ZS-EPMC78302613346718310.3390/genes12010106