Rapid and sensitive detection of “Candidatus Liberibacter asiaticus” by cycleave isothermal and chimeric primer-initiated amplification of nucleic acids
We developed a detection method for “Candidatus Liberibacter asiaticus”, causal agent of citrus huanglongbing, using isothermal and chimeric primer-initiated amplification of nucleic acids combined with cycling probe technology (Cycleave ICAN). With Cycleave ICAN, the reaction was done in one tube in 1 h without the need for electrophoresis, and false positives were not generated. In addition, Cycleave ICAN method was more sensitive than the conventional PCR method. Cycleave ICAN helps shorten the time for the large-scale detection needed to manage huanglongbing.
KeywordsCitrus huanglongbing “Candidatus Liberibacter asiaticus” Citrus psyllid Cycleave isothermal and chimeric primer-initiated amplification of nucleic acids (Cycleave ICAN) Cycling probe technology
We thank Mr. Tsukasa Uehara, Mr. Masaya Maeda, Ms. Eriko Kawatake and Ms. Yuko Murayama for their technical support. We especially thank Mr. Tuyoshi Ooishi for providing citrus psyllids. This research (No.1666) was supported by a grant from Agriculture, Forestry and Fisheries Research Council.
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