PCR Past, Present and Future

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.

Thermovaccination – thermoheliox as a stimulator of the immune response. Kinetics of the synthesis of antibodies and C-reactive protein in coronavirus infection

“…Swabs from the nasopharynx and oropharynx for the detection of coronavirus RNA SARSCoV-2 by polymerase chain reaction (PCR) testing (detection amplifier CFX-96 “REAL TIME” (Bio-Rad, USA)) [ [6] , [7] , [8] ], computed tomography (CT) of lungs and venous blood sampling for the content of immunoglobulins IgG (quantitative) and IgM (quantitative) to coronavirus SARS-CoV-2 according to standard enzyme-linked immunosorbent assay (ELISA) technique (immunochemiluminescent analyzer, Mindray 6000, USA) [ 8 ] were carried out in all patients.…”

Development of RT-PCR Based Diagnosis of SARS-CoV-2

“…Lots of kits have been manufactured throughout the world with some or little variation, thus making it more sensitive, specific and less time consuming. One factor which essential under pandemic situation was a multiple sample analysis in one go [5]. Thus more focus on using portable POC systems, we can imagine the use of micro-fluidics doing concurrent multiple sample analysis.…”

“…Normally the best way to detect any virus from the sample is its isolation and further confirmation by various molecular techniques. But Center for Diseases Control and Prevention (CDC) recommended not to isolate the SARS-CoV-2 as it is a new virus and this practices could be a risky approach and suggested to use patients sample directly for diagnosis [5]. Most of the diagnostic methods are molecular-based hence for diagnosis virus genome study become necessary.…”

Nucleic Acid Testing of SARS-CoV-2

“…In recent years, we have seen the advance of digital PCR (dPCR) as a complementary approach for measuring nucleic acids, a technique that is highly accurate and reproducible when targeting the viral genes of SARS-CoV-2 [ 114 , 115 , 116 , 117 ]. The advantages of digital PCR compared to quantitative PCR include quantification without the need for calibration curves, higher accuracy, and sensitivity that may arise from sub-optimal amplification efficacy because dPCR can detect low amounts of nucleic acid [ 118 , 119 ].…”