Digital PCR as a Highly Sensitive Diagnostic Tool: a Review

Abstract

Nowadays digital PCR (dPCR) is a nucleic acid quantification method widely used in genetic analysis. One of the most significant advantages of dPCR over other methods is the possibility for absolute quantitative determination of genetic material without construction of calibration curves, which allows one to detect even single molecules of nucleic acids, and, hence, early diagnosis of diseases. A specific characteristic of dPCR is the detection of the analyzed biological object in each microreaction, followed by the presentation of the analysis results in a binary system, thereby giving the method name. The key aspects of developing the dPCR method, i.e. from the first devices based on microfluidic chip technology to modern systems capable of measuring a target at a concentration of up to 1 in 100 000 copies were shown in the current work. We analyzed the data on the detection of various pathogens using dPCR, as well as summarized various study results demonstrating the innovativeness of this method “point-of-care”. Both the possibilities of multiplex dPCR analysis and its potential in clinical practice were presented. The review also addresses the issue of the dPCR role in the development of non-invasive methods for oncological diseases to be analyzed. Possible ways of developing dPCR technology were emphasized, including the use as a “point-of-care” systems.