柑橘黄龙病实时荧光PCR鉴定技术研究

[目的]建立一套高效、准确的柑橘黄龙病（HLB）早期鉴定手段。[方法]根据HLBsecE基因（EF164805.1）保守区域设计引物和探针,建立HLB的TaqMan探针实时荧光PCR鉴定技术。[结果]被检测的236个柑橘苗圃样品中,有54个苗圃感染了HLB,检出率为22.9%。引物特异性好。[结论]该检测方法具有快速、灵敏、重复性好等优点,其检测灵敏度达0.01 ng。

Quick Detection of Huanglongbing by Realtime Fluorescent PCR Using TaqMan Probe

[Objective] The aim of study was to optimize an efficient and accurate method to detect HLB.[Method] A real-time fluorescent PCR(RTF-PCR) method was established to detect HLB.Primers and TaqMan probe were designed based on the HLB secE gene.[Result] Large number seedings were detedted by RTF-PCR,the positive percentage was 22.9%(54/236).The primers specificity was good.[Conclusion]The described method was quick,sensitive,stable reproductive and without post-PCR handling of the amplification products.HLB could be detected at low concentration about 0.01 ng.