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Single-Step Recombinase Polymerase Amplification Assay Based on a Paper Chip for Simultaneous Detection of Multiple Foodborne Pathogens.

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A paper chip device-based recombinase polymerase amplification (RPA) method was developed for highly sensitive and selective single-step detection of foodborne pathogens. The diagnostic utility of the device was demonstrated by the reliable detection of E. coli and S.aureus present in spiked milk. This ready-to-use device could be integrated with simple nucleic acid extraction for food pathogen detection in resource-limited settings.

Single-Step Recombinase Polymerase Amplification Assay Based on a Paper Chip for Simultaneous Detection of Multiple Foodborne Pathogens. Read More

Development of Molecular Methods to Detect Macrophomina phaseolina from Strawberry Plants and Soil.

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RPA has been validated on over 200 infected strawberry plants with a diagnostic sensitivity of 93% and a diagnostic specificity of 99%, respectively. Together, this work demonstrates the value of using new approaches to identify loci for detection and provides valuable diagnostic tools that can be used to monitor soil and strawberry plant samples for M. phaseolina.

Development of Molecular Methods to Detect Macrophomina phaseolina from Strawberry Plants and Soil. Read More

A novel recombinase polymerase amplification (RPA) assay for the rapid isothermal detection of Neospora caninum in aborted bovine fetuses

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A recombinase polymerase amplification (RPA) assay combined with lateral flow (LF) strips was developed for the detection of N. caninum. The LF-RPA assay distinguished N. caninum from nine closely related protozoan species. The amplification reaction was completed in about 10 min under isothermal condition. The LF-RPA assay can detect N. caninum DNA at amounts as low as 50 fg.

A novel recombinase polymerase amplification (RPA) assay for the rapid isothermal detection of Neospora caninum in aborted bovine fetuses Read More

Semi-quantitative nucleic acid test with simultaneous isotachophoretic extraction and amplification.

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We demonstrate simultaneous ITP and RPA can consistently detect 5 copies per reaction in buffer and 10 000 copies per milliliter of human serum with no intermediate user steps. We also show preliminary extraction and amplification of DNA from whole blood samples.

Semi-quantitative nucleic acid test with simultaneous isotachophoretic extraction and amplification. Read More

A novel method to detect meat adulteration by recombinase polymerase amplification and SYBR green I

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At the isothermal temperature of 37 °C, RPA specifically identifies duck, chicken, cow, sheep and pig within 30 min of water bath. The RPA amplicons were successfully visualized by adding SG I. Furthermore, RPA can differentiate species of boiled, microwaved, high pressured or fried samples. Finally, using this system, we visually identified 1% pork adulterated in mutton or beef.

A novel method to detect meat adulteration by recombinase polymerase amplification and SYBR green I Read More

Rapid visual detection of cyprinid herpesvirus 2 by recombinase polymerase amplification combined with a lateral flow dipstick.

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Herein, a rapid and convenient detection assay based on RPA with a lateral flow dipstick (LFD) was developed for detecting CyHV‐2. The highly conserved ORF72 of CyHV‐2 was targeted by specific and sensitive primers and probes. The optimized assay takes only 15 min at 38°C using a water bath, with analysis of products by 2% agarose gel electrophoresis within 30

Rapid visual detection of cyprinid herpesvirus 2 by recombinase polymerase amplification combined with a lateral flow dipstick. Read More

Efficacy of Recombinase Polymerase Amplification to Diagnose Trypanosoma cruzi Infection in Dogs with Cardiac Alterations from an Endemic Area of Mexico

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The analytical sensitivity indicated that RPA-LF amplified T. cruzi DNA in samples containing almost equal to one to two parasites per reaction. Serial twofold dilutions of T. cruzi epimastigotes showed that the test had 95% (19/20) repeatability at concentrations of two parasites per reaction. The test showed no cross reactivity with human DNA or other protozoan parasites (Trypanosoma rangeli, Leishmania

Efficacy of Recombinase Polymerase Amplification to Diagnose Trypanosoma cruzi Infection in Dogs with Cardiac Alterations from an Endemic Area of Mexico Read More

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