Advances in improvement strategies of digital nucleic acid amplification for pathogen detection

Digital nucleic acid amplification technologies (dNAATs) have become an emerging platform for pathogen detection. It is not only suitable for the absolute quantitative analysis of target pathogens with very low concentration in samples, but also have strong resistance to complex matrix. Thanks to the rapid development of isothermal amplification and micromachining technology, tremendous progress has been made in the partitioning capabilities, detection range and interpretation accuracy of dNAATs. Herein, the basic principles of digital detection and its advantages in nucleic acid detection is summarized, the types of the reaction system digitization are categorized, a comprehensive discussion on the latest improved design of dNAATs is presented. In the end, the challenges and future directions of the dNAATs developments are outlined. This review provides a comprehensive guidance for the design of dNAATs to improve its distribution efficiency, detection range and cost-effectiveness, thereby contributing to the prevention and control of pathogens. • The digital strategy improves the performance of nucleic acid detection. • Microfluidic technology makes it possible to integrate and miniaturize devices. • The improved scheme of partition reduces the reagent consumption. • The multiplicity of amplification methods reduces the threshold of digital detection. • Artificial intelligence-based recognition algorithm eliminated interference.