Viral diagnostics in the era of digital polymerase chain reaction

Abstract

Unlike quantitative PCR (qPCR), digital PCR (dPCR) achieves sensitive and accurate absolute quantitation of a DNA sample without the need for a standard curve. A single PCR reaction is divided into many separate reactions that each have a positive or negative signal. By applying Poisson statistics, the number of DNA molecules in the original sample is directly calculated from the number of positive and negative reactions. The recent availability of multiple commercial dPCR platforms has led to increased interest in clinical diagnostic applications, such as low viral load detection and low abundance mutant detection, where dPCR could be superior to traditional qPCR.Here we review current literature that demonstrates dPCR’s potential utility in viral diagnostics, particularly through absolute quantification of target DNA sequences and rare mutant allele detection.

Current Role of Genetics in Hematologic Malignancies

“…Digital PCR is a new approach to DNA/ RNA detection and quantification. It is emerging as an alternative to conventional RQ-PCR for quantification and low abundance mutation detection [12]. This is a chip based assay in which several samples can be tested simultaneously.…”

Will droplet digital PCR become the test of choice for detecting and quantifying ocularChlamydia trachomatisinfection? Maybe

“…Current options in the ddPCR market have been reviewed by others including editorial review by Nature Methods. The recent publication of guidelines for the reporting of digital PCR experiments indicates that a previously niche technology is now rapidly entering the mainstream [13][14][15].…”

Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control

“…Real-time quantitative polymerase chain reaction (qPCR) is the gold standard method for sensitive, specific, and quantitative detection of nucleic acids, e.g., for HIV-1 viral load testing, detection of bacterial pathogens, and screening for many other organisms [1][2][3] . During real-time qPCR, primers amplify pathogen DNA in cycles, and a fluorescent signal is generated that is proportional to the amount of amplified DNA in the sample at each cycle.…”