Microfluidic lab-on-a-chip (LOC) technologies have been developed as a promising alternative to traditional central laboratory-based analysis approaches over several decades due to the capability of realizing miniaturized multiphase and multistep reactions. In the field of nucleic acid (NA) diagnosis, digital NA detection (dNAD) as a single-molecular-level detection is greatly attributed to the perfect combination of NA amplification and microfluidic LOC techniques. In this chapter, the

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... e, classification, advances, and application of dNAD will be involved. In particular, the focus will be on chip-based dNAD for giving a deep interpretation of the analysis and evaluation of digital detection. The future prospect of dNAD is also anticipated. It is sure that dNAD by means of microfluidic LOC devices as the promising technique will better serve the ambitious plan of precision medicine through absolute quantitation of NA from individuals. Figure 2. Plate-based chip used for the OpenArray RealTime PCR System. A rectilinear array of 3072 microholes with 320 μm in diameter was fabricated in a stainless steel plate (25×75×0.3 mm). The volume of each hole was approximately 33 nanoliers, and to match the pitch of the wells in a 384-well microplate, the 48 groups of 64 holes are spaced at 4.5 mm. Reprinted with permission from Ref. [43] .