| 四种桉树青枯菌DNA提取方法及PCR检测灵敏度比较 |
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| 引用本文： | 王胜坤,王军,徐大平.四种桉树青枯菌DNA提取方法及PCR检测灵敏度比较[J].中国森林病虫,2007,26(5):4-7. |
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| 作者姓名： | 王胜坤 王军 徐大平 |
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| 作者单位： | 1. 中国林业科学研究院热带林业研究所,广东,广州,510520
2. 华南农业大学林学院,广东,广州,510640 |
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| 基金项目： | 国家科技攻关计划;科技部农业科技成果转化基金 |
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| 摘 要： | 以带有青枯菌Ralstonia solanacearum的桉树组织为材料,采用4种不同的提取方法抽提青枯菌DNA,同时利用青枯菌的特异性引物进行PCR扩增,比较了4种DNA的提取方法及PCR检测的灵敏度。结果表明,煮沸法和热裂解法操作相对简单,但检测灵敏度较低,检测限分别为104CFU/mL和103CFU/mL;简化提取法和试剂盒法检测效果较好,均可检测到102CFU/mL的青枯菌;简化提取法比试剂盒法操作更简单,检测成本低,具有更高的应用价值。
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| 关 键 词： | 青枯菌 DNA提取 PCR检测 灵敏度 |
| 文章编号： | 1671-0886（2007）05-0004-04 |
| 修稿时间： | 2007-02-08 |
Comparison of four different DNA extraction methods of Ralstonia solanacearum in eucalyptus and sensitivity of PCR detection |
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| WANG Sheng-kun,et al..Comparison of four different DNA extraction methods of Ralstonia solanacearum in eucalyptus and sensitivity of PCR detection[J].Forest Pest and Disease,2007,26(5):4-7. |
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| Authors: | WANG Sheng-kun |
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| Institution: | Chinese Academy of Forestry, Research Insti- tute of Tropical Forestry, Guangzhou 510520, China |
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| Abstract: | The DNA of Ralstonia solanacearum was extracted from eucalyptus tissue with four different methods and amplified with specific primers of R.solanacearum.The sensitivities of PCR detection were compared.The results showed that the methods of boiling and cell lysis by heating were easier to operate but had lower sensitivities with 104 CFU/mL and 103 CFU/mL,respectively.DNA-extraction kit method and simple extraction method had better detection effects.The detection limit of both methods was 102 CFU/mL.The simple extraction method with the advantage of easier operation and lower cost may have higher application value. |
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| Keywords: | Ralstonia solanacearum DNA extraction PCR detection sensitivity |
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