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 ).
Read more . . .
Application of Isothermal Amplification Techniques for the Identification of Madurella mycetomatis
Sarah A. Ahmed, Wendy W. J. van de Sande, Marie Desnos-Ollivier, Ahmed H. Fahal, Najwa A. Mhmoud and G. S. de Hoog
Both isothermal amplification techniques show high specificity and sufficient sensitivity to amplify fungal DNA and proved to be appropriate for detection of M. mycetomatis. Diagnostic performance of the techniques is assessed in comparison to conventional PCR using biopsies from eumycetoma patients. RPA is reliable and easy to operate and has a potential to be implemented in mycetoma endemic areas.
Product used: TwistAmp Basic
Detection of Entamoeba histolytica by RPA
Nair G, Rebolledo M, White AC Jr, Crannell Z, Richards-Kortum RR, Pinilla AE, Ramírez JD, López MC, Castellanos-Gonzalez A
Amebiasis is an important cause of diarrheal disease worldwide and has been associated with childhood malnutrition. Here, we report development of a recombinase polymerase amplification (RPA) method to detect E. histolytica specifically. Using visual detection by lateral flow (LF), the test was highly sensitive and specific and could be performed without additional equipment. The availability of this inexpensive, sensitive, and field-applicable diagnostic test could facilitate rapid diagnosis and treatment of amebiasis in endemic regions.
Products used: TwistAmp nfo
Development of RPA Assays for Detection of Orientia tsutsugamushi or Rickettsia typhi
Chien-Chung Chao ,Tatyana Belinskaya, Zhiwen Zhang, Wei-Mei Ching
Sensitive, specific and rapid diagnostic tests for the detection of Orientia tsutsugamushi (O.tsutsugamushi) and Rickettsia typhi (R. typhi), the causative agents of scrub typhus and murine typhus, respectively, are necessary to accurately and promptly diagnose patients and ensure that they receive proper treatment. Recombinase polymerase amplification (RPA) assays using a lateral flow test (RPA-nfo) and real-time fluorescent detection (RPA-exo) were developed targeting the 47-kDa gene of O. tsutsugamushi or 17 kDa gene of R. typhi. The RPA assay was capable of detecting O. tsutsugamushi or R. typhi at levels comparable to that of the quantitative PCR method.
Products used: TwistAmp exo and nfo
A Novel Molecular Test to Diagnose Canine Visceral Leishmaniasis at the Point of Care
A Castellanos-Gonzalez, O.A. Saldarriaga, L. Tartaglino, R. Gacek, E. Temple, H. Sparks, P. C. Melby, B.L Travi
We developed a diagnostic test that uses isothermal recombinase polymerase amplification (RPA) to detect Leishmania infantum. This method was coupled with lateral flow (LF) reading with the naked eye to be adapted as a point-of-care test. The L. infantum RPA-LF had an analytical sensitivity similar to real time-PCR, detecting DNA of 0.1 parasites spiked in dog blood, which was equivalent to 40 parasites/mL. This novel molecular test may have a positive impact in leishmaniasis control programs.
Product used: TwistAmp nfo
Development of reverse transcription RPA assay for avian influenza H5N1 HA gene detection
Nahed Yehia, Abdel-Satar Arafa, Ahmed Abd El Wahed, Ahmed A. El-Sanousi, Manfred Weidmann, Mohamed A. Shalaby
• The developed H5 RT-RPA assay was able to detect one RNA molecule in 10 min.
• H5 RT-RPA had the same sensitivity and specificity as real-time RT-PCR.
• H5 RT-RPA was fast and easy to be operated via portable device.
Development of portable and rapid assay for detection of emerging avian influenza A (H7N9) virus
Ahmed Abd El Wahed, Manfred Weidmann, Frank T. Hufert
• The mobile suitcase laboratory is easy to use in low-resource settings.
• The Diagnostics-in-a-Suitcase depends on using cold-chain-independent reagents.
• The electricity was supplied from a motor vehicle battery or solar panel battery.
• The H7N9 RT-RPA assays provided results within 10 min.
Product used: TwistAmp exo-RT
Breed-Specific Detection of Mangalica Meat in Food Products
R. Szántó-Egész, A. Jánosi, A. Mohr, G. Szalai, E. Koppányné Szabó, A. Micsinai, R. Sipos, J. Rátky, I. Anton, A. Zsolnai
A fast and reliable diagnostic system has been developed for the detection of Mangalica meat in foods. This qualitative test is based on a recombinase polymerase amplification, which can be performed in the field, in situ, where it may be necessary to determine Mangalica content in food products at once.
Product used: TwistAmp nfo
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). "