| 程保平, 彭埃天* , 宋晓兵, 凌金锋, 陈霞.三种PCR方法检测柑橘黄龙病菌的效果比较[J].植物保护,2014,40(5):106-110. |
| 三种PCR方法检测柑橘黄龙病菌的效果比较 |
| Comparison of citrus huanglongbing molecular detection methods |
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| DOI： |
| 中文关键词: 柑橘黄龙病 常规PCR 巢式PCR 实时荧光定量PCR 比较研究 |
| 英文关键词:Candidatus Liberibacter asiaticus conventional PCR nested PCR real-time quantitative PCR comparative research |
| 基金项目:广东省农业科技推广专项资金重大技术研究项目(201201127); 广东省现代农业产业技术体系建设专项(粤农［2009］380号); 广东省农业科学院院长基金项目(201305); 科技创新人才出国培养计划(粤农科［2013］74号) |
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| 中文摘要: |
| 为了比较常规PCR、巢式PCR和实时荧光定量PCR方法在大田检测中对柑橘黄龙病(Huanglongbing, HLB)的检测效果, 首先比较了3种检测方法对柑橘黄龙病菌检测的灵敏度, 结果发现：3种检测方法的灵敏度依次为常规PCR<巢式PCR<实时荧光定量PCR。运用3种检测方法对广东5个柑橘品种上的189个黄龙病疑似病样进行检测, 结果发现：黄龙病检出率依次为常规PCR<巢式PCR<实时荧光定量PCR。研究表明：常规PCR适合以较低成本大规模检测黄龙病; 实时荧光定量PCR具有最大的检测灵敏度; 巢式PCR检测技术同时具有前两者的一些优点, 但操作较复杂, 适合技术熟练的研究者使用。 |
| 英文摘要: |
| Detection sensitivity and detection rate of conventional PCR, nested PCR and real-time quantitative PCR were compared for detection of citrus huanglongbing. The results showed that the detection sensitivity was ranked from high to low as real-time quantitative PCR>nested PCR>conventional PCR. Then total 189 suspected citrus samples from five different cultivars with symptom of HLB in orchard of Guangdong were amplified and confirmed by the three methods. The results indicated that the detection rate was ranked from high to low as real-time quantitative PCR>nested PCR>conventional PCR. This research indicated that conventional PCR, du to its low cost, could be used in large scale diagnosis of HLB disease, real-time quantitative PCR had the highest detection sensitivity, and nested PCR had the advantages of the other two, but its experimental operation was more complex. |
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