Abstract Citrus Huanglongbing (HLB) is one of the most devastating diseases in the citrus industry and is caused primarily by ‘ Candidatus Liberibacter asiaticus’ ( C Las), a phloem-restricted gram-negative bacterium transmitted via citrus psyllids in the field. Precise C Las 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 C Las 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 C Las five-copy nrd B gene (β-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 C Las 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 C Las in field conditions when coupled with simplified crude DNA extraction method, providing an alternative approach for on-site C Las detection in the context of HLB.
Fu et al. (Wed,) studied this question.
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