Infectious diseases seriously endanger human health. Rapid and accurate detection of pathogens is the key to effective treatment and precise prevention and control of infectious diseases. Traditional pathogen testing techniques cover a limited variety of microorganisms, making it difficult to meet clinical needs. Metagenomic Next Generation Sequencing (mNGS) theoretically can simultaneously detect all known gene sequences of pathogens, greatly improving the clinical capacity of diagnosing and treating patients with severe, complicated, rare, and emerging pathogen infections. In the context of individualized health needs, the self-built testing of new technologies in laboratories can continuously improve the level of clinical diagnosis and treatment. Due to the high starting point of research and development and the complex operation process of the mNGS technology platform, this study reaches a consensus on the experimental procedure, assay validation, quality control, and report review of the mNGS-LDT in the fields of clinical testing, infection, critical care, and in vitro diagnosis to ensure the appropriate application of the technology and guarantee patient safety and proposes standardized requirements and suggestions.