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Droplet digital PCR of viral DNA/RNA, current progress, challenges, and future perspectives
Author(s) -
Kojabad Amir Asri,
Farzanehpour Mahdieh,
Galeh Hadi Esmaeili Gouvarchin,
Dorostkar Ruhollah,
Jafarpour Ali,
Bolandian Masoumeh,
Nodooshan Majid Mirzaei
Publication year - 2021
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.26846
Subject(s) - digital polymerase chain reaction , polymerase chain reaction , dna , rna , computational biology , real time polymerase chain reaction , microbiology and biotechnology , biology , virology , genetics , gene
High‐throughput droplet‐based digital PCR (ddPCR) is a refinement of the conventional polymerase chain reaction (PCR) methods. In ddPCR, DNA/RNA is encapsulated stochastically inside the microdroplets as reaction chambers. A small percentage of the reaction chamber contains one or fewer copies of the DNA or RNA. After PCR amplification, concentrations are determined based on the proportion of nonfluorescent partitions through the Poisson distribution. Some of the main features of ddPCR include high sensitivity and specificity, absolute quantification without a standard curve, high reproducibility, good tolerance to PCR inhibitor, and high efficacy compared to conventional molecular methods. These advantages make ddPCR a valuable addition to the virologist's toolbox. The following review outlines the recent technological advances in ddPCR methods and their applications in viral identification.