Achieving precise dual detection: One-tube reverse transcription-recombinase aided amplification (RT-RAA) combined with lateral flow strip (LFS) assay for RNA and DNA target genes from pepper mild mottle virus and Colletotrichum species in crude plant samples
文献类型： 外文期刊
作者： Cao, Yuhao 1 ; Yan, Dankan 1 ; Zhou, Huijie 1 ; Han, Kelei 1 ; Wan, Qionglian 1 ; Peng, Jiejun 1 ; Zheng, Hongying 1 ; Lin, Lin 1 ; Yan, Fei 1 ; Song, Xuemei 1 ;
作者机构： 1.Ningbo Univ, Inst Plant Virol, State Key Lab Managing Biot & Chem Threats Qual &, Ningbo 315211, Peoples R China
2.Ningbo Univ, Inst Plant Virol, Key Lab Biotechnol Plant Protect MARA, Key Lab Green Plant Protect Zhejiang Prov, Ningbo 315211, Peoples R China
3.Anhui Acad Agr Sci, Inst Plant Protect & Agroprod Safety, Hefei 230031, Peoples R China
关键词： Anthracnose; Dual RT RAA; LFS; On-site detection; PMMoV
期刊名称：TALANTA （ 影响因子：6.1； 五年影响因子：5.5 ）
ISSN： 0039-9140
年卷期： 2025 年 281 卷
页码：
收录情况： SCI
摘要： Ensuring the detection sensitivity of both RNA-derived and DNA-derived target genes in a single reaction has posed a significant challenge for on-site detection of plant pathogens. This challenge was addressed by developing a one-tube dual RT-RAA assay combined with LFS for the rapid on-site detection of pepper mild mottle virus (PMMoV) and four Colletotrichum species causing anthracnose in Solanaceous crops. By testing four different combinations of primer groups, two combinations were precisely adjusted within the dual RT-RAA system to balance amplification efficiency and maintain consistent levels of amplification in crude plant samples. Utilizing commercially accessible small-scale equipment and following a streamlined optimization strategy, the assay achieved a limit of detection of 0.32 copies/mu L of target genes in the reaction. Importantly, it demonstrated no cross-reactivity with other plant pathogens, thereby affirming the high sensitivity and specificity of the developed dual RT-RAA-LFS detection assay. Moreover, the entire process took only 25 min from sample collection to the visible presentation of results. The assay was validated with 60 field samples and 10 seed samples, producing results consistent with reverse transcription quantitative polymerase chain reaction (RT-qPCR). Notably, it successfully detected PMMoV in systemic leaves without visible symptoms three days post-inoculation, underscoring its effectiveness in early disease detection. This streamlined strategy offers a valuable approach for rapid, low-cost, and highly sensitive on-site simultaneous detection of RNA genome-contained PMMoV and DNA genome-contained Colletotrichum species.
- 相关文献
作者其他论文 更多>>
-
Effects of water-soluble protein of turtle shell on the quality and in vitro digestibility of rice noodles
作者：Ren, Mengying;Lu, Jianfeng;Wang, Hong;Song, Yuan;Lin, Lin;Ren, Mengying;Lu, Jianfeng;Wang, Hong;Song, Yuan;Lin, Lin;Li, Zhengrong;Lai, Nianyue;Zhou, Beibei;Yang, Li;Xiang, Xudong;Han, Xiaomiao
关键词：Turtle shell extract; Water-soluble protein; Rice noodles; Eating quality; Gelatinization; Digestibility
-
Synergistic effects of alternating magnetic field and sodium tripolyphosphate on functional properties of myofibrillar proteins in low-salt systems
作者：Li, Qiang;Xia, Lizhi;Lu, Jianfeng;Song, Yuan;Yang, Mengru;Lin, Lin;Zheng, Zhi;Li, Qiang;Xia, Lizhi;Lu, Jianfeng;Song, Yuan;Yang, Mengru;Lin, Lin;Zheng, Zhi;Zhou, Beibei;Miao, Wenhua;Zheng, Bin
关键词：Myofibrillar protein; Alternating magnetic field; Sodium tripolyphosphate; Gel properties; Low salt
-
The nanovirus U2 protein suppresses RNA silencing via three conserved cysteine residues
作者：Yan, Dankan;Chen, Jianping;Yan, Dankan;Han, Kelei;Lu, Yuwen;Peng, Jiejun;Rao, Shaofei;Wu, Guanwei;Chen, Jianping;Zheng, Hongying;Yan, Fei;Yan, Dankan;Han, Kelei;Liu, Yong
关键词：cysteine; milk vetch dwarf virus; Nanovirus; U2 protein; viral suppressor of RNA silencing
-
Identification and complete genome sequence of iris potyvirus A, which causes dwarfing and foliar chlorosis with mosaic or mottle disease symptoms on lily (Lilium lancifolium Thunb.) in China
作者：Wang, Fang;Yan, Dankan;Han, Kelei;Ma, Chao;Chen, Ying;Gao, Zhengliang;Bao, Xianxun;Li, Cheng
关键词：
-
Complete genome sequence of triticum yellow stripe virus, a new polerovirus infecting wheat (triticum aestivum) in China
作者：Yan, Dankan;Han, Kelei;Chen, Ying;Ma, Chao;Hu, Shuzhen;Zhao, Wei;Wang, Fang
关键词：
-
Pepper mild mottle virus coat protein interacts with pepper chloroplast outer envelope membrane protein OMP24 to inhibit antiviral immunity in plants
作者：Han, Kelei;Zheng, Hongying;Zhou, Huijie;Jia, Zhaoxing;Zhai, Yushan;Wu, Jian;Lu, Yuwen;Wu, Guanwei;Rao, Shaofei;Chen, Jianping;Peng, Jiejun;Yan, Fei;Han, Kelei;Yan, Dankan;Qi, Rende
关键词：
-
Chloroplast clustering around the nucleus induced by OMP24 overexpression unexpectedly promoted PSTVd infection in Nicotiana benthamiana
作者：Han, Kelei;Jia, Zhaoxing;Zhang, Yuhong;Zhou, Huijie;Bu, Shan;Chen, Jianping;Yan, Fei;Wu, Jian;Han, Kelei;Jia, Zhaoxing;Zhang, Yuhong;Zhou, Huijie;Bu, Shan;Chen, Jianping;Wu, Jian;Yan, Dankan;Qi, Rende;Yan, Fei;Wu, Jian
关键词：chloroplast outer membrane protein 24; H2O2; perinuclear chloroplast clustering; potato spindle tuber viroid
