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Isothermal polymerase chain reaction


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​​What is isothermal PCR?


​Isothermal PCR is a branch of PCR in which DNA amplification occurs at a constant temperature instead of cycling through several temperatures. A number of isothermal technologies have been developed, which employ different enzymes and biochemical techniques to enable separation and replication of double stranded DNA at the same temperature.


How does isothermal PCR work?


Although the core concept of denaturing, extending, and annealing DNA to replicate it remains relatively standard, it is done very differently compared to traditional PCR, and each isothermal technique uses unique reagents and protocols. For example, while traditional PCR uses heat to destabilize the bonds between 2 DNA strands, isothermal PCR uses enzymes called helicase to biochemically break apart DNA bonds at a much lower temperature. By developing special enzymes and primers, the typical steps in replicating DNA can all be performed at the same temperature.


Why is isothermal PCR useful?


Because isothermal PCR does not require instrumentation dedicated to thermally cycle the sample, the complexity, size, and cost of isothermal devices can potentially be much lower than traditional PCR machines. There is therefore significant interest in developing such devices for use in point-of-care and limited resource settings.