Recombinase polymerase amplification (RPA) is a single tube, isothermal alternative to the polymerase chain reaction (PCR).[1] By adding a reverse transcriptase enzyme to an RPA reaction it can detect RNA as well as DNA, without the need for a separate step to produce cDNA,.[2][3][4] Because it is isothermal, RPA can use much simpler equipment than PCR, which requires a thermal cycler. Operating best at temperatures of 37–42 °C and still working, albeit more slowly, at room temperature means RPA reactions can in theory be run quickly simply by holding a tube. This makes RPA an excellent candidate for developing low-cost, rapid, point-of-care molecular tests. An international quality assessment of molecular detection of Rift Valley fever virus performed as well as the best RT-PCR tests, detecting less concentrated samples missed by some PCR tests and an RT-LAMP test.[5]
RPA was developed and launched by TwistDx Ltd. (formerly known as ASM Scientific Ltd), a biotechnology company based in Cambridge, UK.
^Piepenburg, Olaf; Williams, Colin H.; Stemple, Derek L.; Armes, Niall A. (2006). "DNA Detection Using Recombination Proteins". PLOS Biology. 4 (7): e204. doi:10.1371/journal.pbio.0040204. PMC 1475771. PMID 16756388.
^Euler, Milena; Wang, Yongjie; Nentwich, Oliver; Piepenburg, Olaf; Hufert, Frank T.; Weidmann, Manfred (2012). "Recombinase polymerase amplification assay for rapid detection of Rift Valley fever virus". Journal of Clinical Virology. 54 (4): 308–12. doi:10.1016/j.jcv.2012.05.006. PMID 22683006.
^Amer, H.M.; Abd El Wahed, A.; Shalaby, M.A.; Almajhdi, F.N.; Hufert, F.T.; Weidmann, M. (2013). "A new approach for diagnosis of bovine coronavirus using a reverse transcription recombinase polymerase amplification assay". Journal of Virological Methods. 193 (2): 337–40. doi:10.1016/j.jviromet.2013.06.027. PMC 7113639. PMID 23811231.
^Abd El Wahed, Ahmed; El-Deeb, Ayman; El-Tholoth, Mohamed; Abd El Kader, Hanaa; Ahmed, Abeer; Hassan, Sayed; Hoffmann, Bernd; Haas, Bernd; Shalaby, Mohamed A.; Hufert, Frank T.; Weidmann, Manfred (2013). Meng, Xiang-Jin (ed.). "A Portable Reverse Transcription Recombinase Polymerase Amplification Assay for Rapid Detection of Foot-and-Mouth Disease Virus". PLOS ONE. 8 (8): e71642. Bibcode:2013PLoSO...871642A. doi:10.1371/journal.pone.0071642. PMC 3748043. PMID 23977101.
^Escadafal, Camille; Paweska, Janusz T.; Grobbelaar, Antoinette; Le Roux, Chantel; Bouloy, Michèle; Patel, Pranav; Teichmann, Anette; Donoso-Mantke, Oliver; Niedrig, Matthias (2013). De Silva, Aravinda M (ed.). "International External Quality Assessment of Molecular Detection of Rift Valley Fever Virus". PLOS Neglected Tropical Diseases. 7 (5): e2244. doi:10.1371/journal.pntd.0002244. PMC 3662703. PMID 23717706.
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