重组酶聚合酶扩增
斑点钩端螺旋体
环介导等温扩增
病菌
生物
DNA
微生物学
基因
生物化学
作者
Rong Lei,Pinshan Wu,Limei Li,Qunxing Huang,Jingyi Wang,Di Zhang,Mingfu Li,Naizhong Chen,Xinyi Wang
标识
DOI:10.1016/j.snb.2021.129874
摘要
Sensitive and rapid detection of plant pathogens, particularly for on-site diagnosis during custom inspections, is essential to prevent and control the spread of these pathogens. To address the need, a combined assay for detecting plant pathogens was developed by using gold nanoparticle-enhanced dynamic microcantilever (MCL) biosensing and isothermal recombinase polymerase amplification (RPA). In this study, we used Leptosphaeria maculans (L. maculans), an aggressive fungus responsible for major losses in oilseed rape yield worldwide, as the target pathogen to demonstrate the merits of the combined RPA and MCL strategy (named as the RPA-MCL assay). The results indicated that the sensitivity of the RPA-MCL assay is higher than that of the fluorescence RPA assay, with the detection limit of only one copy of L. maculans DNA. In the practical assay, the newly developed RPA-MCL method was found to detect 57 ppm L. maculans genomic DNA in the oilseed rape seeds genomic DNA sample. Considering the high sensitivity and specificity of this strategy, we envision that the proposed RPA-MCL assay could have wide applications in nucleic acid diagnostics for on-site plant pathogen detection and species identification.
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