| 赵培娅,高玉霞,卢占军.两组探针和SYBR Green染料qPCR检测柑橘黄龙病的灵敏度研究[J].中国南方果树,2016,45(2): |
| 两组探针和SYBR Green染料qPCR检测柑橘黄龙病的灵敏度研究 |
| Sensitivity comparison of two TaqMan probes and SYBR Green qPCR method on detecting Citrus Huanglongbing |
| 投稿时间：2016-02-23 修订日期：2016-03-08 |
| DOI： |
| 中文关键词: 柑橘黄龙病 TaqMan水解探针 SYBR Green染料法 实时荧光定量PCR 灵敏度 |
| 英文关键词:Citrus Huanglongbing Taq Man probes SYBR Green method Quantitative Real-time PCR Sensitivity |
| 基金项目:国家自然科学基金（31560602）；江西省科技计划重大项目（20152ACF60003）；江西省科技厅青年科学基金（20131542040011）；江西省教育厅重点项目（GJJ12564）；江西省自然科学基金（20122BAB204024）；江西省自然科学基金（20131542040011）；江西省科技计划重大项目（20143ACF60012）；江西省教育厅科技项目（GJJ14661）资助。 |
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| 中文摘要: |
| 本研究把柑橘黄龙病（Citrus Huanglongbing，HLB）阳性核酸样品进行梯度稀释后，分别采用HLBp、CQULAP两组TaqMan水解探针法和以rpl基因为目的片段的SYBR Green染料法进行HLB实时荧光定量PCR(Quantitative Real-time PCR, qPCR)灵敏度检测研究，结果发现HLBp探针法可以从105倍稀释的阳性叶片核酸样品中检测到病原菌（病菌拷贝数约为60个），高于CQULAP探针法和SYBR Green染料法；将三种qPCR检测体系用于赣州安远县、寻乌县和赣县送检的60份田间样品检测，结果HLBp探针阳性率为86.66%，CQULAP探针阳性率为60%，以rpl基因为目的片段的SYBR Green染料法检测阳性率为68.33%。 |
| 英文摘要: |
| Three Quantitative Real-time PCR (qPCR) methods were compared using TaqMan probe HLBp, TaqMan probe CQULAP and SYBR Green targeted rpl gene, for diagnosis of Citrus Huanglongbing (HLB) disease. The sensitivity of three protocols were tested for detecting ‘Candidatus Liberibacter asiaticus’ in a series of gradient diluted total DNA extracted of infected citrus leaves with HLB. The results showed TaqMan probe HLBp could detect pathogen in 105 diluted total DNA extracts sample (about 60 pathogen copy number), which sensitivity was higher than TaqMan probe CQULAP and SYBR Green method. To further confirm sensitivity of three qPCR protocols, 60 leaf samples from orchards in Anyuan county, Xunwu county, Ganxian county were detected by TaqMan probe HLBp, TaqMan probe CQULAP and SYBR Green targeted rpl gene, respectively. Assays showed TaqMan probe HLBp has the highest positive rate(86.66%), TaqMan probe CQULAP and SYBR Green method showed positive rate 60% and 68.33%, respectively. |
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