Rapid detection of foot-and-mouth disease virus using reverse transcription recombinase polymerase amplification combined with a lateral flow dipstick

Wang, Hongmei; Zhao, Guimin; Hou, Peili; Yu, Li; He, Cheng-qiang; He, Hongbin

doi:10.1016/j.jviromet.2018.07.011

Cited by 21 publications

(14 citation statements)

References 27 publications

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“…RPA, another isothermal amplification assay, requires only two primers, is carried out at a lower temperature (39°C) and takes shorter time (<30 min). Due to its advantages compared to other isothermal amplification methods, the RPA method has been widely used for rapid detection of multiple pathogens that infected with human beings and animals (Chen et al., ; Ma, Zeng, Huang, et al., ; Moore & Jaykus, ; Vasileva Wand, Bonney, Watson, Graham, & Hewson, ; Wang, Zhao, et al., ). For point‐of‐care diagnostics in remote area, RPA method is of great interest due to its portability, short turnaround time and potential for construction of a mobile laboratory.…”

Section: Discussionmentioning

confidence: 99%

Point‐of‐care diagnostic assay for rapid detection of porcine deltacoronavirus using the recombinase polymerase amplification method

Zeng

Huang

et al. 2019

Transbound Emerg Dis

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Summary Porcine deltacoronavirus (PDCoV) has emerged and spread throughout the porcine industry in many countries over the last 6 years. PDCoV caused watery diarrhoea, vomiting and dehydration in newborn piglets. A sensitive diagnostic method would be beneficial to the prevention and control of PDCoV infection. Recombinase polymerase amplification (RPA) is an isothermal amplification method which has been widely used for virus detection. A probe‐based reverse transcription RPA (RT‐RPA) assay was developed for real‐time detection of PDCoV. The amplification can be finished in 20 min and fluorescence monitoring was performed by a portable device. The lowest detection limit of the PDCoV RT‐RPA assay was 100 copies of RNA molecules per reaction; moreover, the RT‐RPA assay had no cross‐reaction with other common swine viruses. The clinical performance of the RT‐RPA assay was evaluated using 108 clinical samples (54 intestine specimens and 54 faecal swab specimens). The coincidence rate of the detection results for clinical samples between RT‐RPA and RT‐qPCR was 97.2%. In summary, the real‐time RT‐RPA assay offers a promising alternative to RT‐qPCR for point‐of‐care detection of PDCoV.

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Section: Discussionmentioning

confidence: 99%

Point‐of‐care diagnostic assay for rapid detection of porcine deltacoronavirus using the recombinase polymerase amplification method

Zeng

Huang

et al. 2019

Transbound Emerg Dis

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“…One promising solution is the RPA assay. Due to its convenience and portability, the RPA technique has been used for rapid detection of various pathogens, such as foot-and-mouth disease virus [29], Zika virus [30], human norovirus [24] and pathogenic bacteria [31]. Up to date, there is only one report about the RPA assay for the laboratory animal health monitoring [32].…”

Section: Discussionmentioning

confidence: 99%

Development and evaluation of a broadly reactive reverse transcription recombinase polymerase amplification assay for rapid detection of murine norovirus

Zeng

Feng

et al. 2018

BMC Vet Res

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BackgroundMurine norovirus (MNV) is recognized as the most prevalent viral pathogen in captive mouse colonies. The rapid detection assay for MNV would be a useful tool for monitoring and preventing MNV infection. A recombinase polymerase amplification (RPA) assay was established in this study to provide a solution for rapid and sensitive detection of MNV.ResultsThe detection limit of the RT-RPA assay for the detection of MNV was 1 × 102 copies of RNA molecules per reaction. The assay was specific since there was no cross-reaction with other common murine viruses. In addition, the broad reactivity of the RT-RPA assay was validated using the synthesized template carrying seven point mutations among several MNV strains. The MNV RT-RPA assay could detect as few as 1 × 102 copies of the mutant per reaction, suggesting the assay could be broadly reactive against a large diversity of MNV strains. Forty eight clinical samples including 16 gastric tissue specimens, 16 cecal tissue specimens and 16 fecal specimens were tested for the validation of the new developed RT-RPA assay. The detection results of RT-RPA and RT-qPCR for clinical samples were very similar, except that a gastric tissue sample which was positive by RT-qPCR, with a RNA titer of 27 copies, was negative by RT-RPA.ConclusionsA broadly reactive RT-RPA assay was successfully established for MNV detection.

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“…Although it has been proved that the pan-specific real-time RT-RPA (rRT-RPA) and RPA-LFD technique can provide rapid and accurate diagnosis of FMDV [ 12 , 18 ], they are difficult to distinguish the serotypes of FMDV for the rRT-RPA assay. The VP1, a surface exposed-capsid protein, take a pivotal role in the antigenicity as a major viral antigen, and plays an important role in pathogenicity of FMDV as its binding to viral receptors of host cells.…”

Section: Discussionmentioning

confidence: 99%

“…RT-RPA primers and probes specific for serotypes O, A or Asia-1 of FMDV were designed against the consensus sequence for this region and were synthesized by Sangon Biotech, respectively. RPA primers/probes were synthesized and labeled as described in previous study [ 18 ]. Oligonucleotide sequences of RPA primers and probes of specific serotype A, Asia-1, or O of FMDV are listed in Table 1 (accession numbers KT968663, EF149010 and HQ009509, respectively).…”

Section: Methodsmentioning

confidence: 99%

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Development and evaluation of serotype-specific recombinase polymerase amplification combined with lateral flow dipstick assays for the diagnosis of foot-and-mouth disease virus serotype A, O and Asia1

et al. 2018

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BackgroundFoot-and-mouth disease (FMD) caused by foot-and-mouth disease virus (FMDV) is one of the most highly infectious diseases in livestock, and leads to huge economic losses. Early diagnosis and rapid differentiation of FMDV serotype is therefore integral to the prevention and control of FMD. In this study, a series of serotype-specific reverse transcription recombinase polymerase amplification assays combined with lateral flow dipstick (RPA-LFD) were establish to differentiate FMDV serotypes A, O or Asia 1, respectively.ResultsThe serotype-specific primers and probes of RPA-LFD were designed to target conserved regions of the FMDV VP1 gene sequence, and three primer and probe sets of serotype-specific RPA-LFD were selected for amplification of FMDV serotypes A, O or Asia 1, respectively. Following incubation at 38 °C for 20 min, the RPA amplification products could be visualized by LFD. Analytical sensitivity of the RPA assay was then determined with ten-fold serial dilutions of RNA of VP1 gene and the recombinant vector respectively containing VP1 gene from FMDV serotypes A, O or Asia1, the detection limits of these assays were 3 copies of plasmid DNA or 50 copies of viral RNA per reaction. Moreover, the specificity of the assay was assessed, and there was no cross reactions with other viruses leading to bovine vesicular lesions. Furthermore, 126 clinical samples were respectively detected with RPA-LFD and real-time PCR (rPCR), there was 98.41% concordance between the two assays, and two samples were positive by RPA-LFD but negative in rPCR, these were confirmed as FMDV-positive through viral isolation in BHK-21 cells. It showed that RPA-LFD assay was more sensitive than the rPCR method in this study.ConclusionThe development of serotype-specific RPA-LFD assay provides a rapid, sensitive, and specific method for differentiation of FMDV serotype A, O or Asia1, respectively. It is possible that the serotype-specific RPA-LFD assay may be used as a integral protocol for field detection of FMDV.

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Rapid detection of foot-and-mouth disease virus using reverse transcription recombinase polymerase amplification combined with a lateral flow dipstick

Cited by 21 publications

References 27 publications

Point‐of‐care diagnostic assay for rapid detection of porcine deltacoronavirus using the recombinase polymerase amplification method

Point‐of‐care diagnostic assay for rapid detection of porcine deltacoronavirus using the recombinase polymerase amplification method

Development and evaluation of a broadly reactive reverse transcription recombinase polymerase amplification assay for rapid detection of murine norovirus

Development and evaluation of serotype-specific recombinase polymerase amplification combined with lateral flow dipstick assays for the diagnosis of foot-and-mouth disease virus serotype A, O and Asia1

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