Development of a recombinase-aided amplification assay for detection of orf virus

Wang, Yong; Cui, Yongqiu; Yu, Zhaorong; Li, Yeqiu; Bai, Caixia; Sun, Peng; Zhu, Wen

doi:10.1016/j.jviromet.2020.113861

Cited by 16 publications

(11 citation statements)

References 18 publications

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“…Thus far, RAA has been widely applied to detect pathogens, including virus (Qi et al 2019;Wang et al 2019bWang et al , 2020c, parasites (Li and Yang, 2019;Ni et al 2019;Zhang et al 2019b), and other causative agents. It has also been employed in single-nucleotide polymorphism analysis (Fu et al 2019b), plant quarantine (Tang et al 2019), and food testing (Miao et al 2019).…”

Section: Discussionmentioning

confidence: 99%

Recombinase-aided amplification combined with lateral flow dipstick for the rapid detection of Amphidinium carterae

Zhang

Liu

et al. 2021

J Appl Phycol

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In the context of global climate change, the frequency and duration of harmful algal blooms (HABs) due to eutrophic coastal waters have increased. HABs can cause marine ecological imbalance, massive socio-economic damage, and potential public health problems. Therefore, accurate and specific techniques that can rapidly detect harmful algae are critical to expedite early monitoring and prediction of HABs. Although some methods based on amplification have previously been described, limitations associated with limited popularity and high costs still exist. In this study, the partial sequence of nuclear ribosomal DNA from Amphinidium carterae was selected as the target region. The biotin-labeled recombinase-aided amplification (RAA) products and the 6-carboxyfluorescein-labeled probe were specifically conjugated, which formed double-labeled RAA products analyzed by lateral flow dipstick (LFD). A rapid RAA-LFD assay aimed at visually detecting A. carterae was established and optimized. The specificity and sensitivity of RAA-LFD were determined. The nucleic acid test with other control algal species yielded negative results, which indicated that RAA-LFD assay can specifically detect A. carterae. The detection limit of RAA-LFD assay for A. carterae was 8.49 ng μL −1 genomic DNA and 1 cell mL −1 . The whole detection process of RAA-LFD could be completed within 50 min. In conclusion, the proposed RAA-LFD assay may be promising for the efficient, specific, and rapid detection of A. carterae in monitoring efforts.

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Section: Discussionmentioning

confidence: 99%

Recombinase-aided amplification combined with lateral flow dipstick for the rapid detection of Amphidinium carterae

Zhang

Liu

et al. 2021

J Appl Phycol

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“…Some virulent strains may cause severe symptoms at a low titer and, thus, a sensitive detection method is indispensable in such situations. SYBR Green I-based assays are more rapid, can be implemented more simply, generally enable detection in a single step, and produce results that are consistent with those of other methods, which reflects the greater stability of SYBR Green I-based methods [ 39 , 40 ]. The model exemplified in this study can be extended to the differential diagnosis of other mixed viral infections, which highlights its general significance and applicability to multiple areas of veterinary medicine [ 41 ].…”

Section: Discussionmentioning

confidence: 99%

Simultaneous detection of duck circovirus and novel goose parvovirus via SYBR green I-based duplex real-time polymerase chain reaction analysis

Wang

Cui

et al. 2020

Molecular and Cellular Probes

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Beak atrophy and dwarfism syndrome (BADS) is commonly caused by co-infection with duck circovirus (DuCV) and novel goose parvovirus (NGPV). Therefore, concurrent detection of both viruses is important for monitoring and limiting BADS, although such a diagnostic test has not been reported. In this study, we developed a duplex, SYBR Green I-based real-time polymerase chain reaction (PCR) assay to enable the simultaneous detection of DuCV and NGPV. The assay readily distinguished between the two viruses, based on their different melting temperatures (Tm), where the Tm for DuCV was 80 °C and that for NGPV was 84.5 °C. Other non-target duck viruses that were tested did not show melting peaks. The detection limit of the duplex assay was 10 1 copies/μL for both viruses. This method exhibited high repeatability and reproducibility, and both the inter-assay and intra-assay variation coefficients were <1.6%. Thirty-one fecal samples were collected for clinical testing using real-time PCR analysis, and the results were confirmed using sequencing. The rate of co-infection was 6.5%, which was consistent with the sequencing results. This duplex real-time PCR assay offers advantages over other tests, such as rapid, sensitive, specific, and reliable detection of both viruses in a single sample, which enables the quantitative detection of DuCV and NGPV in clinical samples. Using this test may be instrumental in reducing the incidence of BADS and the associated economic losses in the duck and goose industries.

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“…Recombinase-aided amplification (RAA) assay is a new isothermal amplification technology for pathogens detection with the advantages of rapidity, simplicity, and low cost ( Chen et al, 2018 ; Duan et al, 2018 ; Yan et al, 2018 ; Wang et al, 2020 ; Ruichen Lv et al, 2022 ). This amplification process only requires approximately 15–30 min at 39°C and has been successfully applied for detection of African Swine Fever Virus ( Fan et al, 2020 ), Klebsiella pneumoniae ( Hou et al, 2021 ), human norovirus GII.4 ( Qin et al, 2021 ), duck circovirus ( Li et al, 2021 ), Trypanosoma evansi ( Li et al, 2020 ), etc.…”

Section: Introductionmentioning

confidence: 99%

Development of a Lateral Flow Strip-Based Recombinase-Aided Amplification for Active Chlamydia psittaci Infection

Jiao

et al. 2022

Front. Microbiol.

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Chlamydia psittaci is the causative agent of psittacosis, a worldwide zoonotic disease. A rapid, specific, and sensitive diagnostic assay would be benefit for C. psittaci infection control. In this study, an assay combining recombinase-aided amplification and a lateral flow strip (RAA-LF) for the detection of active C. psittaci infection was developed. The RAA-LF assay targeted the CPSIT_RS02830 gene of C. psittaci and could be accomplished in 15 min at a single temperature (39°C). The analytical sensitivity of the assay was as low as 1 × 100 copies/μl and no cross-reaction with some other intracellular pathogens was observed. Moreover, all feces samples from mice infected with C. psittaci at day-1 post-infection were positive in the RAA-LF assay. In conclusion, the RAA-LF assay provides a convenient, rapid, specific and sensitive method for detection of active C. psittaci infection and it is also suitable for C. psittaci detection in field.

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Development of a recombinase-aided amplification assay for detection of orf virus

Cited by 16 publications

References 18 publications

Recombinase-aided amplification combined with lateral flow dipstick for the rapid detection of Amphidinium carterae

Recombinase-aided amplification combined with lateral flow dipstick for the rapid detection of Amphidinium carterae

Simultaneous detection of duck circovirus and novel goose parvovirus via SYBR green I-based duplex real-time polymerase chain reaction analysis

Development of a Lateral Flow Strip-Based Recombinase-Aided Amplification for Active Chlamydia psittaci Infection

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