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Review
. 2020 Oct;69(4):317-325.
doi: 10.2144/btn-2020-0057. Epub 2020 Aug 20.

PCR past, present and future

Hanliang Zhu  1 , Haoqing Zhang  1 , Ying Xu  1 , Soňa Laššáková  2 , Marie Korabečná  2 , Pavel Neužil  1   3   4
Affiliations
Review

PCR past, present and future

Hanliang Zhu et al. Biotechniques. 2020 Oct.

Abstract

PCR has become one of the most valuable techniques currently used in bioscience, diagnostics and forensic science. Here we review the history of PCR development and the technologies that have evolved from the original PCR method. Currently, there are two main areas of PCR utilization in bioscience: high-throughput PCR systems and microfluidics-based PCR devices for point-of-care (POC) applications. We also discuss the commercialization of these techniques and conclude with a look into their modifications and use in innovative areas of biomedicine. For example, real-time reverse transcription PCR is the gold standard for SARS-CoV-2 diagnoses. It could also be used for POC applications, being a key component of the sample-to-answer system.

Keywords: COVID-19; PCR; RNA virus diagnoses; digital PCR; microfluidics; point-of-care diagnostics; portable systems; reverse transcription PCR.

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Figures

Figure 1.
Figure 1.. The development of the PCR system and its applications.
(A) A prototype PCR thermal cycler developed by Cetus Corporation in 1986, which is the first model embedding the software cycling controller in the thermal cycling block. (B) Space-domain PCR. (C) Time-domain PCR. (D) Use of anchored allele-specific probes and labeled primers for the colorimetric detection of mutations in the HBB gene. (E) One of the first applications of multiplex PCR, the detection of deletions in the DMD gene in patients affected by Duchenne muscular dystrophy. (F) A recent version of the multiplex PCR with fluorescent-labeled primers and separation of amplicons using capillary electrophoresis currently used in routine forensic analysis. (G) The use of PCR for the amplification of alleles of a multiallelic minisatellite locus, silver staining of amplicons in a polyacrylamide gel, and the comparison of their lengths with an allele marker, a ladder. Reproduced with permission from [4–7].
Figure 2.
Figure 2.. Current examples of commercially available techniques: quantitative PCR, droplet-based digital PCR, crystal digital PCR (cdPCR), PCR, bridge PCR for next-generation sequencing and sequencing of mRNA from individual cells using microfluidics.
(A) A comparison of end-point PCR, qPCR and ddPCR. (B) Schematic representation of the principle of solid phase bridge DNA amplification. (C) Different techniques for splitting of samples. (D) Crystal droplet PCR – formation of droplet crystals. (E) PCR and droplet-based library generation for single-cell RNA sequencing. ddPCR: Droplet-based digital PCR; qPCR: Quantitative PCR. (A) Reproduced with permission from [33]. (B) Reproduced with permission from [34], © Elsevier BV (2014).
Figure 3.
Figure 3.. Applications of microfluidics into massively parallel and handheld point-of-care systems.
(A) Chip-based integrated real-time reverse transcription PCR platform for the analysis of the immunomagnetic exosomal RNA. (B) Droplet-based quantitative PCR for a single cell to mRNA purification and gene expression analysis. (C) Chip-based digital RT-PCR for absolutequantification of mRNA in single cells. (D) Droplet-based dPCR for miRNA quantitation assay. (E) Paper-based LAMP system made by polydimethylsiloxane for molecular diagnostics. (F) Forensic science, DNA profiles on a chip. (G) BioFire, detection of bacteria and viruses on a chip. Reproduced with permission from [38,50,68,69].
See this image and copyright information in PMC

References

    1. Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc. Natl Acad. Sci. USA 74(12), 5463–5467 (1977). - PMC - PubMed
    1. Mullis K, Faloona F, Scharf S, Saiki R, Horn G, Erlich H. Specific enzymatic amplification of DNA in vitro: the polymerase chain reaction. Cold Spring Harb. Symp. Quant. Biol. 51(Pt. 1), 263–273 (1986). - PubMed

    2. •• The first description of PCR appeared after the patent application written in 1985 (patent approved in July 1987).

    1. Saiki RK, Scharf S, Faloona F. et al. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle-cell anemia. Science 230(4732), 1350–1354 (1985). - PubMed
    1. Hurd C, Cavanagh G, Schuh A, Ouwehand W, Metcalfe P. Genotyping for platelet-specific antigens: techniques for the detection of single nucleotide polymorphisms. Vox Sang. 83(1), 1–12 (2002). - PubMed
    1. Chamberlain JS, Gibbs RA, Rainer JE, Nguyen PN, Thomas C. Deletion screening of the Duchenne muscular dystrophy locus via multiplex DNA amplification. Nucleic Acids Res. 16(23), 11141–11156 (1988). - PMC - PubMed

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