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 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(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 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.展开更多
基金supported by the National Key R&D Program of China(2021YFD1400800)the Innovation Research 2035 Pilot Plan of Southwest University(SWU-XDZD22002)+2 种基金the Special Fund for Youth Team of Southwest University(SWU-XJLJ202310)the Natural Sciences Foundation of Chongqing,China(CSTB2024 NSCQ-MSX1133)the Southwest University Introduction of Talents Program(SWU120023).
文摘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(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 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.
