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A Revolution in DNA Detection


We have enclosed some publications below you may find of interest.


    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.

    Read more . . .

    NB The formulation of TwistAmp kits is substantially different to that used in this publication (different proteins, different crowding agent, different protein ratios etc) to favour faster amplification of shorter amplicons.

  • A Fully Integrated Lab-on-a-Disc for Nucleic Acid Analysis of Food-borne Pathogens

    TwistFlow® Salmonella tests integrated into a centrifugal microfluidic device that performs DNA extraction, amplification and detection onto a single disc. DNA extraction to readout took 30 minutes with sensitivities of 10 CFU/ml of PBS and 100 CFU/ml of milk.

    Read more . . . 

  • Rapid detection of Plasmodium falciparum with isothermal recombinase polymerase amplification

    Rapid detection of Plasmodium falciparum with isothermal recombinase polymerase amplification and lateral flow analysis.

    A TwistAmp nfo-based test for the 18S rRNA gene of P. falciparum detected as few as four parasites after <10 minutes of amplification. Time to result, including lateral flow detection was 15 minutes.

    Read more . . .

  • Rapid Molecular Assays for the Detection of Yellow Fever Virus in Low-Resource Settings

    RPA assays were developed to detect Yellow Fever Virus on three different platforms (real-time with or without microfluidic semi-automated system and lateral-flow assay). The assay was able to detect 20 different YFV strains with no cross-reactions with closely related viruses. The RPA assay proved to be a robust, portable method with a low detection limit (<21 genome equivalent copies per reaction) and rapid processing time (<20 min). Results from real-time RPA field testing were comparable to results obtained in the laboratory, demonstrating the method is suitable for YFV detection in low-resource settings.

    Read more . . .

  • Multiplex isothermal solid-phase RPA for DNA-based detection of three bacterial pathogens

    Solid-phase RPA to simultaneously amplify and detect  Neisseria gonorrhoeae, Salmonella enterica,  methicillin-resistant Staphylococcus aureus (MRSA) and a control plasmid in <20 minutes. The assay can detect 10 CFU of S. enterica and MRSA and 100 CFU of N. gonorrhoeae.

    Read more . . . 

  • A Nucleic Acid Test to Diagnose Cryptosporidiosis: Lab Assessment in Animal and Patient Specimens

    RPA detection of Cryptosporidium species in DNA extracted from stool samples. TwistAmp nfo based assays were more sensitive than RT-PCR and only needed 30 minutes of amplification time. Using a simple paper and plastic device, lateral flow strips were one to two orders of magnitude more sensitive than detection of products by gel electrophoresis.

    Read more . . . 

  • Isothermal RPA Assay Applied to the Detection of Group B Streptococci

    Isothermal Recombinase Polymerase Amplification Assay Applied to the Detection of Group B Streptococci inVaginal / Anal Samples.

    Group B streptococcal infections are the leading cause of sepsis and meningitis in newborns. A rapid and reliable method for thedetection of this pathogen at the time of delivery is needed for the early treatment of neonates. A RPA assay showed clinical sensitivity of 96% and specificity of 100% as compared to RT-PCR, but in <20 minutes.

    Read more . . . 

  • Immunosorbent assay as a DNA amplification-detection strategy for food analysis

    Recombinase polymerase and enzyme-linked immunosorbent assay as a DNA amplification-detection strategy forfood analysis.

    The first publication showing that RPA can be combined with ELISA. The RPA-ELISA combination is proposed for amplification at a low, constant temperature (40°C) in a short time (40min), for the hybridisation of labelled products to specific 5'-biotinylated probes/streptavidin in coated microtiter plates at room temperature, and for detection by colorimetric immunoassay. RPA-ELISA was applied to screen common safety threats in foodstuffs, such as allergens (hazelnut, peanut, soybean, tomato, and maize), genetically modified organisms (P35S and TNOS), pathogenic bacteria (Salmonella sp. and Cronobacter sp.), and fungi (Fusarium sp.). Satisfactory sensitivity and reproducibility results were achieved for all the targets. The RPA-ELISA technique does away with thermocycling and provides a suitable sensitive, specific, and cost-effective method for routine applications, and proves particularly useful for resource-limited settings.

    Read more . . .

  • Sensitive and Rapid Detection of Chlamydia trachomatis by RPA Directly from Urine Samples

    A 20 minute, lateral flow-based, RPA test to detect Chlamydia trachomatis directly from urine, with no purification step. The test described works with 10% urine, has a specificity of 100% (58/58 negatives) and a sensitivity of 83% (10/12 positives detected) based on a small clinical trial. The whole procedure is fairly simple and does not require specific machinery, making it potentially applicable in point-of-care settings.

    Read more . . .

  • RT RPA Assay for the Detection of Middle East Respiratory Syndrome Coronavirus

    An RT-RPA reaction that detects MERS-CoV in 3-7 minutes, with the same sensitivity as RT-PCR is shown running on a portable, Twista-like, device. The assay is described as being “ideal for rapid mobile molecular MERS-CoV monitoring in acute patients and may also facilitate the search for the animal reservoir of MERS-CoV”

    Read more . . . 

  • Creating Biocompatible Oil-Water Interfaces without Synthesis

    Creating Biocompatible Oil-Water Interfaces without Synthesis: Direct Interactions between Primary Amines and Carboxylated Perfluorocarbon Surfactants.

    A novel method for generating biocompatible droplet surfaces capable of supporting  picoliter-scale RPA reactions. This paper shows that RPA can be used in a digital, droplet form.

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  • A Portable RT-RPA Assay for Rapid Detection of Foot-and-Mouth Disease Virus

    Foot-and-mouth disease (FMD) causes serious economic damage and is a serious infectious threat to livestock. Early diagnosis of FMD helps to diminish its impact by improving outbreak management. This study describes the development of a real-time reverse transcription RPA assay for detection of FMD virus. The RPA assay took 4-10 minutes and detected all seven FMDV serotypes. The FMDV RT-RPA assay was successfully used in the field in Egypt during a recent FMD outbreak. Clinical sensitivity was 98% (vs 100% for RT-PCR), but FMDV RT-RPA was much quicker and easier to handle in the field than RT-PCR. The authors conclude that RT-RPA could be easily implemented to perform diagnostics at quarantine stations or farms for rapid spo-of-infection detection.

    Read more . . .

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