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Multiplex lateral flow immunoassay for simultaneous detection and specific discrimination of Xoo and Xoc in rice plants

文献类型： 外文期刊

作者： Zhang, Cui 1 ; Zhang, Xi 1 ; Liu, Nairu 1 ; Dong, Jie 1 ; Mao, Weijia 1 ; Liu, Zhaoli 3 ; Zhou, Xueping 1 ; Wu, Jianxiang 1 ;

作者机构： 1.Zhejiang Univ, Inst Biotechnol, State Key Lab Rice Biol & Breeding, Zhejiang Key Lab Biol & Ecol Regulat Crop Pathogen, Hangzhou 310058, Peoples R China

2.Zhejiang Univ, Hainan Inst, Sanya 572025, Peoples R China

3.Peking Univ, Coll Environm Sci & Engn, Beijing, Peoples R China

4.Chinese Acad Agr Sci, Inst Plant Protect, State Key Lab Biol Plant Dis & Insect Pests, Beijing 100193, Peoples R China

关键词： Multiplex lateral flow immunoassay; Simultaneous detection; AuNPs; Xoo; Xoc

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 292 卷

页码：

收录情况： SCI

摘要： Xanthomonas oryzae pv. oryzae (Xoo) and Xanthomonas oryzae pv. oryzicola (Xoc) are two important rice bacterial pathogens, causing serious losses of global rice yield every year. Effective management of rice bacterial diseases needs timely and accurate detection and quarantine techniques. Herein, a novel Au nanoparticles-based multiplex lateral flow immunoassay (AuNP-mLFIA) was developed for simultaneous detection and specific discrimination of Xoo and Xoc in rice leaf and seed samples. Four anti-Xoo mAbs, four anti-Xoc mAbs, and ten broad-spectrum mAbs towards both Xoo and Xoc were first obtained via hybridoma cell technique. Through checkerboard assays, pairwise mAbs 12H1/19C9 against Xoo and self-paired mAb 18D12 against Xoc were selected to guarantee the highest sensitivity and specificity of the assay. Under the optimal conditions, the AuNP-mLFIA achieved a visual limit of detection (LOD) as low as 7.8 x 10(3) CFU mL(-1) for Xoo and 1.6 x 10(4) CFU mL(-1) for Xoc with no cross-reactivity. In addition, the assay was successfully applied to monitor Xoo and Xoc in 40 field-collected rice leaf and seed samples and the detection results of this assay agreed well with conventional PCR. Therefore, this developed AuNP-mLFIA was sensitive, rapid, and accurate for simultaneous detection of Xoo and Xoc. This study also provides guidelines for the screening of paired mAbs to establish a double-antibody sandwich (DAS)-mLFIA.

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