Development of a real-time reverse transcription recombinase polymerase amplification assay for rapid detection of spring viremia of carp virus

Feng, Cong; Zeng, Fanwen; Wu, Miaoli; Wang, Jingjing; Huang, Bihong; Wang, Yingying; Wang, Qing; Zhang, Shouquan; Ma, Lei; Guo, Pengju; Zeng, Weiwei

doi:10.1016/j.mcp.2019.101494

Cited by 13 publications

(9 citation statements)

References 34 publications

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“…RPA, a simple, rapid, and low cost without the need of sophisticated equipment or specialized skill, is of great application. For instance, RPA is used to detect genotype III grass carp reovirus [14] , shrimp hemocyte iridescent virus [15] and spring viremia of carp virus [16] . Previous reports described that RPA was more sensitive, rapid and accurate than traditional PCR [17] .…”

Section: Introductionmentioning

confidence: 99%

Rapid and sensitive detection of pathogenic Elizabethkingia miricola in black spotted frog by RPA-LFD and fluorescent probe-based RPA

Qiao

Zhang²,

Chang³

et al. 2022

Fish and Shellfish Immunology Reports

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Section: Introductionmentioning

confidence: 99%

Rapid and sensitive detection of pathogenic Elizabethkingia miricola in black spotted frog by RPA-LFD and fluorescent probe-based RPA

Qiao

Zhang²,

Chang³

et al. 2022

Fish and Shellfish Immunology Reports

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“…A lateral flow dipstick RPA method to detect Salmonella in milk samples yielded a LOD of 10 2 CFU/ml (Kim et al, 2014). The RPA has been recently applied for the rapid detection of spring viraemia of carp virus (Cong et al, 2019) and murine norovirus with LODs of 10 2 virion copies/reaction.…”

Section: Discussionmentioning

confidence: 99%

“…It is a simple, specific, sensitive, rapid and economical method that can be adapted to point-of-care detection of clinical samples (Daher et al, 2016;Piepenburg et al, 2006). RPA methods have been successfully developed for the rapid detection and diagnosis of several important pathogens including viruses (Abd et al, 2013;Cong et al, 2019;Wang et al, 2019), bacteria (Gao et al, 2018;Geng et al, 2019;Hansen et al, 2016) and parasites (Chao et al, 2015;Kersting et al, 2014b). The detection of amplification products in real time using a fluorescent dye makes this method adaptable to real-time PCR thermocyclers, and data can be displayed using the real-time thermocycler software as progress curves with threshold times and fluorescence intensities.…”

Section: Loop-mediated Isothermal Amplification Assays Have Been Recementioning

confidence: 99%

Development of a real‐time recombinase polymerase amplification assay for rapid detection of Aeromonas hydrophila

Wang

et al. 2020

Journal of Fish Diseases

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The genus Aeromonas is widespread in environmental habitats such as water and soil and is pathogen of cold-blooded animals and mammals including humans. Aeromonads are common inhabitants detected in most types of food, for example in dairy products (4%), vegetables (26%-41%), and meats and poultry (3%-70%), with the largest numbers recorded from shellfish (31%) and fish (72%) (Janda & Abbott, 2010). Aeromonas hydrophila is the most common of the Aeromonad found in freshwater throughout the world, and infections

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“…RPA is emerging as an innovative isothermal PCR alternative for rapid detection of diseases. RPA assays are most often developed to enable real-time fluorescence detection, with detection of several viral shrimp diseases described [18][19][20]29,30] with reported sensitivities ranging from 4 to 100 molecular copies of the pathogens. However, fluorescent reading instruments require additional power requirements and expense that can preclude uptake for lowresource implementation.…”

Section: Plos Onementioning

confidence: 99%

Rapid sample preparation and low-resource molecular detection of hepatopancreatic parvoviruses (HPV) by recombinase polymerase amplification lateral flow detection assay in shrimps (Fenneropenaeus merguiensis)

et al. 2022

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Background Viral diseases are a major problem in shrimp aquaculture facilities as these diseases reduce growth rates, which inevitably lead to production and profit losses. Hepatopancreatic parvoviruses (HPV) are common diseases in shrimp that appear to be associated with high or low levels of replication in specific genetic lineages. Selective breeding may result in resistance to HPV and improved body traits such as body weight, meat yield and shrimp colour, facilitating shrimp farming. HPV virus titre is commonly determined by quantitative PCR (qPCR), which is a time-consuming method requiring laboratory equipment unsuitable for field implementation. The aim of this study was to develop a simple, robust, rapid and reliable method to detect HPV in low-resource environments. Methods We developed a rapid shrimp HPV test that uses (1) a simple three-step sample preparation protocol, followed by (2) isothermal recombinase polymerase amplification (RPA) and lateral flow strip detection (LFD). Analytical sensitivity testing was performed in a background banana shrimp sample matrix, and retrospective testing of Fenneropenaeus merguiensis hepatopancreas tissues (n = 33) with known qPCR viral titres was used to determine diagnostic sensitivity and specificity. Results The rapid shrimp HPV test could detect as little as 35 genome-equivalent copies per reaction in homogenized F. merguiensis banana shrimp. Retrospective testing of stored tissues (n = 33) indicated 100% diagnostic sensitivity (95% confidence interval, CI: 86–100%) and 100% specificity (95% CI: 66–100%) for detection of HPV. Conclusion The rapid shrimp HPV test could be completed in only 40 minutes, and required only homogenization pestles, some pipettors, and a small heating block for single temperature incubation at 39°C. Critically, our procedure eliminated the time-consuming purification of nucleic acids from samples and when combined with RPA-LFD offers a user-friendly HPV detection format that can potentially be performed on-site. Our approach represents a major step forward in the development of a simple and sensitive end-point method for quick determination of unfavourable HPV virus numbers in shrimp, and has great potential to advance on-site management of shrimps in aquaculture.

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Development of a real-time reverse transcription recombinase polymerase amplification assay for rapid detection of spring viremia of carp virus

Cited by 13 publications

References 34 publications

Rapid and sensitive detection of pathogenic Elizabethkingia miricola in black spotted frog by RPA-LFD and fluorescent probe-based RPA

Rapid and sensitive detection of pathogenic Elizabethkingia miricola in black spotted frog by RPA-LFD and fluorescent probe-based RPA

Development of a real‐time recombinase polymerase amplification assay for rapid detection of Aeromonas hydrophila

Rapid sample preparation and low-resource molecular detection of hepatopancreatic parvoviruses (HPV) by recombinase polymerase amplification lateral flow detection assay in shrimps (Fenneropenaeus merguiensis)

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