Field applicable all-in-one kit developed for rapid detection of 'Candidatus Liberibacter asiaticus' in plants and psyllids

文献类型： 外文期刊

第一作者： Fu, Shimin

作者： Fu, Shimin;Wan, Hongcen;Duan, Qiuyuan;Wang, Xuefeng;Zhou, Changyong;Lou, Binghai;Bai, Ziqin

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关键词： Citrus Huanglongbing; CLas detection kit; Field application; NrdB

期刊名称：PHYTOPATHOLOGY RESEARCH （ 影响因子：3.5； 五年影响因子：3.9 ）

ISSN： 2096-5362

年卷期： 2025 年 7 卷 1 期

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收录情况： SCI

摘要： Citrus Huanglongbing (HLB) is one of the most devastating diseases in the citrus industry and is caused primarily by 'Candidatus Liberibacter asiaticus' (CLas), a phloem-restricted gram-negative bacterium transmitted via citrus psyllids in the field. Precise CLas detection is crucial for HLB control, particularly during extensive surveys in new emerging regions. Unfortunately, the lack of a practical on-site detection method for CLas due to the limited specificity of immunostrips and the costive non-user friendliness approaches prevent the effective disease control. In this study, a probe-based recombinase polymerase amplification (RPA) targeting the CLas five-copy nrdB gene (beta-subunit of ribonucleotide reductase, RNR) was developed and evaluated for its specificity, sensitivity, and reliability. To enhance user-friendliness and facilitate widely use, an All-in-one kit was premade as freeze-dried pellets in individual tubes, containing all necessary components except DNA template. The applicability of the All-in-one kit for CLas detection was confirmed using plant or psyllid crude DNA extracts. Collectively, the All-in-one kit offers a specific, sensitive, rapid, cost-effective, and practical alternative for diagnosing CLas in field conditions when coupled with simplified crude DNA extraction method, providing an alternative approach for on-site CLas detection in the context of HLB.

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