Every year more and more scientists are finding out that RPA really works. Nearly 50 publications have been written in the last 4 years and the rate is going up as people discover the benefits of RPA. See the publications on this page to give you an idea of how you could use RPA for experiements that just aren't possible with PCR.
DNA DETECTION USING RECOMBINATION PROTEINS
The original RPA publication...
DNA amplification is essential to most nucleic acid testing strategies, but established techniques require sophisticated equipment or complex experimental procedures, and their uptake outside specialised laboratories has been limited.
Users interested in using RPA to amplify MRSA DNA should use the oligonucleotides described in our more recent publication ( http://www.ncbi.nlm.nih.gov/pubmed/24972080 ).
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Detection and Characterization of Viral Species/Subspecies Using Isothermal RPA Assays.
"In this paper, the complete procedures to set up an RPA assay, applied to detection of RNA (Potato virus Y, Potyvirus) and DNA (Wheat dwarf virus, Mastrevirus) viruses, are described. The proposed procedure allows developing species- or subspecies-specific detection assay."
Product used: TwistAmp Basic kit
Rapid & sensitive detection of antibiotic resistance on a programmable digital microfluidic platform
"We describe a rapid and sensitive detection system for analysis of DNA containing the blaCTX-M-15 gene using isothermal DNA amplification by recombinase polymerase amplification (RPA) on a digital microfluidic platform; active matrix electrowetting-on-dielectric (AM-EWOD)."
Product used: TwistAmp exo kits
A field-applicable RPA assay for rapid detection of Mycoplasma capricolum subsp. capripneumoniae.
"Contagious caprine pleuropneumonia (CCPP) is a highly contagious disease caused by Mycoplasma capricolum subsp. capripneumoniae (Mccp) that affects goats in Africa and Asia...We report a rapid, specific and sensitive assay employing isothermal DNA amplification using recombinase polymerase amplification (RPA) for detection of Mccp."
Rapid and specific detection of Yam mosaic virus by reverse-transcription RPA
•RT-RPA test for specific detection of YMV detects as low as 14 pg/μl of viral RNA.
•First RT-RPA test for a root and tuber crop virus.
•RT-RPA is more suitable than RT-PCR for routine screening of YMV.
Yam mosaic virus (YMV; genus Potyvirus) is considered to cause the most economically important viral disease of yams (Dioscorea spp.) in West Africa which is the dominant region for yam production globally... The RT-exoRPA assay has, however, several advantages over the RT-PCR; positive samples can be detected in less than 30 min, and amplification only requires a single incubation temperature (optimum 37 °C). "
Recombinase Polymerase Amplification Assay for Rapid Diagnostics of Dengue Infection
Real-time RT-PCR analysis is not suitable for on-site screening since mobile devices are large, expensive, and complex. In this study, two RT-recombinase polymerase amplification (RT-RPA) assays were developed to detect DENV1-4.
Real-time & label-free ring-resonator monitoring of solid-phase recombinase polymerase amplification
• Development of a robust and automated solid-phase amplification platform for optical detection of dsDNA targets.
• Real-time and label-free monitoring of isothermal solid-phase amplification was achieved using ring resonators in combination with recombinase polymerase amplification.
• The amplification of a 144-bp target was successfully demonstrated with a limit of detection of 7.8-10−13 M (6-105 copies in 50 -L).
Development of portable and rapid assay for the detection of emerging avian influenza A (H7N9) virus
• The mobile suitcase laboratory is easy to use in low resource settings.
• The Diagnostics-in-a-Suitcase depends on using cold chain independent reagents.
• Electricity is supplied from a motor vehicle battery or solar panel battery.
• The H7N9 RT-RPA assays provided results within 10 minutes.
Centrifugal step emulsification for absolute quantification of nucleic acids by digital droplet RPA
The centrifugal microfluidic droplet generation was used to perform the first digital droplet recombinase polymerase amplification (ddRPA). It was used for absolute quantification of Listerias monocytogenes DNA concentration standards with a total analysis time below 30 min – about a quarter of the time it would take for ddPCR.
Product used: TwistAmp exo+ Listeria
Development of a Quantitative RPA Assay with an Internal Positive Control
A video protocol for developing a real-time recombinase polymerase amplification assay to quantify initial concentration of DNA samples using either a thermal cycler or a microscope and stage heater. Also described is the development of an internal positive control. Scripts are provided for processing raw real-time fluorescence data.
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Solid-phase RPA on microfluidic DVDs
Standard un-modified DVD discs and commercial drives are used for low-cost DNA detection. DNA primers were printed in a microarray format on the polycarbonate surfaces of DVDs with integrated control spots to guarantee the absence of false-negatives and false-positives. The solid-phase amplification assay, including the washing protocols and development reaction, was performed by the dispensation of solutions through the inlet and by controlling the flow-movement by DVD drive centrifugation. The final disc with reaction products was inserted into a DVD player and microarray images were captured and automatically processed.
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Integrated Quantum Dot Barcode Smartphone Optical Device for Wireless Multiplexed Diagnostics
RPA is combined with quantum dot barcodes to allow multiplex detection of pathogens with a smartphone. RPA boosted the sensitivity of pathogen detection in a situation where PCR was not possible.
Product used: TwistAmp® Basic
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