Polymerase chain reaction (PCR) is a powerful technique used to amplify specific fragments
of DNA sequences of interest through repeated thermal cycling. For amplification to be
effective, the PCR machine must be able to achieve and maintain certain temperatures for
specified durations. These temperature dependent steps include, typically, between 90-94°C
for the denaturation of the double-stranded (ds) DNA, 50-70°C for annealing of the primers
to the single stranded (ss) DNA, and 70-75°C for enzymatic extension of the primers.
In general, a traditional PCR takes around 2 hours, though this does not include sample
preparation, subsequent analysis, and or the desired number of amplification cycles. Though
the denaturation and renaturation steps can be performed in just a few minutes, PCR cycle
times are ultimately limited by the elongation step which is ultimately dependent on the
length of the amplified product.
While conventional PCR can provide sufficient quantities of material for detection even at
the single copy level, fast reporting of the results can be severely hindered. Rapid reporting
of PCR results is particularly important in applications such as the detection of pathogens, for
example the SARS-CoV-2 virus, or for real-time molecular forecasting before a sensitive
surgical procedure.
A variation of the technique, called rapid or fast PCR, uses specialized enzymes and reaction
conditions to amplify DNA in a quicker, timely manner, when compared to traditional PCR.
In general, rapid PCR techniques overcome time limitations by removing the need for
excessive thermal cycling. Instead, rapid PCR utilized systems that contain small capillary-
channeled vessels with a lower thermal capacitance which allows for faster heat transfer.
A rapid PCR test is so quick and sensitive that it has been used for point of care testing, and
can be performed in a medical office or pharmacy using desktop machines without the need
to send the sample to a highly complex laboratory. These tests are used, more simply, to
confirm the presence or absence of viral RNA in a sample where extensive DNA
amplification is not necessary. Rapid PCR tests are often compared to rapid antigen tests due
to their quick testing times. Importantly though, rapid PCR tests are more accurate than
antigen tests as they identify specific genetic material from a pathogen, and not just specific
sugars or proteins on an antigens surface.
Outside of point of care diagnostic testing, rapid PCR has various applications and has been
used in forensics, paternity testing, to diagnose genetic disorders as well as cancer.
Additionally, it can be used to study gene expression, regulation, and function, or as a quality
control step for pharmaceuticals and biologics. Though rapid PCR tests offer fast results and
reduce the time required for amplification, PCR product yields will be decreased, and results
may have lower sensitivity or accuracy compared to standard PCR. Rapid PCR tests, similar
to traditional PCR, may suffer from false-positive and false-negative results, so a secondary
test is often needed to confirm results.
Comparison of Rapid or Fast PCR vs standardized PCR. Many variations of fast PCR are differentiated
by an enzyme-modified polymerase, increasing its affinity for single-stranded DNA. This shortens the
time of the procedure considerably. Figure made in BioRender.
Polymerase chain reaction (PCR) is a
