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Tests of recombinant plasmid-spiked serum samples revealed that RPA and real-time PCR had the same diagnostic rate. The RPA assay, which is simple, cost-effective, and fast, is a promising alternative to real-time PCR for ASFV detection.
• This is the first study to use RPA to detect AbHV and RGNNV. • Reaction can be finished at 37 °C in 20 min; time can be further reduced to 5 min for high viral load sample. • The detection limits are 100 viral DNA copies per reaction for both viruses. • Detection methods for both viruses have good
We developed a new surface-enhanced Raman scattering (SERS)-based lateral flow (LF) strip biosensor combined with recombinase polymerase amplification (RPA) for simultaneous detection of Listeria monocytogenes and Salmonella enterica serotype Enteritidis. AuMBA@Ag core–shell nanoparticles were used in this SERS-LF.
•LAMP and RPA methods were developed for the detection of L. monocytogenes. •Performance of the new methods was compared against two reference real-time PCR methods. •A very low limit of detection, with high confidence, was obtained for both methodologies. •Both methodologies proved reliable alternatives to conventional culture methods.
The amplification product was visualized by the lateral flow (LF) strip within 5 min using the specific probe added to the RPA reaction system. The sensitivity of the established assay was 10 times higher than that of nested PCR with a lower detection limit of 0.1 oocyst per reaction, and there was no cross-reactivity with other closely related protozoan species.
The assay is based on the amplification of an imp gene fragment, highly specific for this pathogen. Moreover, we demonstrated that this assay can be used on phloem sap directly squeezed from infected plant material and that the pathogen, can be detected by RPA in real time mode or by a lateral flow device.
RT-exo RPA analysis of the infected plants using the primer/probe indicated that the virus could be detected from leaves, stems, petals, pollen, primary roots and secondary roots.
A low cost thin-film transistor (TFT) nanoribbon (NR) sensor has been developed for rapid real-time detection of DNA amplification using an isothermal Recombinase Polymerase Amplification (RPA) method.
The panel included RPAs for Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, and Enterococcus faecalis. All five RPAs required reaction times of under 12 minutes to reach their lower limit of detection of 100 genomes per reaction or less, and did not cross-react with high concentrations of non-target bacterial genomic DNA.
In this study, an assay was developed and evaluated for the detection of banana bunchy top virus (BBTV) in infected banana plants. Three oligonucleotide primer pairs were designed from the replicase initiator protein gene sequences of BBTV to function both in RPA as well as in polymerase chain reaction (PCR).
