Nucleic Acid Detection Revolution: A Sharp Alternative to PCR
It's been thirty years since the birth of PCR. From classical PCR, real-time quantitative PCR to the current digital PCR, the technology is constantly changing but has never faded from our view.
RPA (Recombinase Polymerase Amplification) is said to be an alternative to PCR for nucleic acid detection. TwistDx Inc, a British company, has developed TwistAmp? Nucleic Acid Amplification products based on this technology, which are capable of detecting single molecules of nucleic acids at room temperature in less than 15 minutes. The technology requires very little hardware equipment and is particularly suitable for use in in vitro diagnostics, veterinary medicine, food safety, biosecurity, agriculture and other fields.
What's so great about RPA technology
Conventional PCR must go through three steps: denaturation, annealing, and extension, and a PCR instrument is essentially a temperature-controlled device that can be used to raise or lower the temperature of the reaction. This can undoubtedly greatly speed up the PCR. In addition, since no temperature control equipment is required, RPA can truly realize portable and rapid nucleic acid detection.
It is reported that the RPA assay is highly sensitive and can amplify trace nucleic acid (especially DNA) templates to detectable levels, yielding about 1012 amplification products from a single template molecule. RPA also eliminates the need for complex sample processing, making it suitable for field assays where nucleic acids cannot be extracted.
RPA amplifies both DNA and RNA and eliminates the need for additional cDNA synthesis steps. You can not only perform endpoint detection of the amplification product, but also monitor the amplification process in real time, and even read the results via test strips (Lateral Flow Chromatography Test Strips LFD).
Currently, the amplicon length of the TwistAmp?kit is within 500bp. However, specially optimized RPA amplification can also generate longer amplification products, or slow down the amplification (for easier quantification), or even perform the reaction at a lower temperature.