Abstract
The infection of Bombyx mori nucleopolyhedrovirus (BmNPV) is one of the main causes of economic losses in sericulture. Thus, it is essential to establish rapid and effective method for BmNPV detection. In the present study, we have developed a recombinase-aided amplification (RAA) to amplify the BmNPV genomic DNA at 37 °C within 30 min, and achieved a rapid detection method by coupling with a lateral flow dipstick (LFD). The RAA-LFD method had a satisfactory detection limit of 6 copies/μL of recombinant plasmid pMD19-T-IE1, and BmNPV infection of silkworm can be detected 12 h post-infection. This method was highly specific for BmNPV, and without cross-reactivity to other silkworm pathogens. In contrast to conventional polymerase chain reaction (PCR), the RAA-LFD assay showed higher sensitivity, cost-saving, and especially is apt to on-site detection of BmNPV infection in the sericulture production.
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Acknowledgements
We are grateful to all who provided the means for us to access free software, which we have used and cited in this article. We would also like to thank all partners and laboratory staff for kindly help and criticism.
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This work was funded by the China Agriculture Research System of MOF and MARA. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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Runpeng Wang performed the main experiments and drafted the manuscript; Zhongyuan Shen were responsible for experimental design; Sheng Xu, Erjun Wei, and Ping He provided technical support and conducted data analysis. Yiling Zhang, Qiang Wang, and Xudong Tang revised the manuscript. All authors reviewed the manuscript.
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Wang, R., Xu, S., Wei, E. et al. Recombinase-aided amplification coupled with lateral flow dipstick for efficient and accurate detection of Bombyx mori nucleopolyhedrovirus. Folia Microbiol (2023). https://doi.org/10.1007/s12223-023-01102-7
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DOI: https://doi.org/10.1007/s12223-023-01102-7