Targeted next-generation sequencing (tNGS): An upcoming application for pathogen identification in clinical diagnosis

Rapid and efficient detection of pathogens is a goal for clinical laboratories. Traditional methods such as post-culture identification, smear microscopy, antigen-antibody detection, and polymerase chain reaction cannot perform hundreds of high-throughput tests simultaneously. With the development of genetic engineering, next-generation sequencing technology has gradually been introduced into clinical testing. The new technology of targeted next-generation sequencing (tNGS) has the advantages of high sensitivity, high efficiency, and relatively low cost. This review of articles on tNGS published in the past two decades summarizes tNGS workflow and clinical applications, compares it with traditional culture and metagenomic next-generation sequencing (mNGS), and highlights its role in detecting multiple pathogens in mixed infections and drug-resistance genes. It also elaborates on the latest guidelines and the quality control process. By highlighting the practical clinical use of tNGS, it offers insights for optimizing the detection of multiple pathogens in polymicrobial infections to enhance diagnostic accuracy and efficiency.