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| 18srDNA为内参照的Real-Time-PCR方法检测半夏病毒 | |
| 引用本文： | 何煜波,陈集双,陈海敏,冯俊丽,高必达.18srDNA为内参照的Real-Time-PCR方法检测半夏病毒[J].湖南师范大学自然科学学报,2006,29(3):103-106. |
| 作者姓名： | 何煜波 陈集双 陈海敏 冯俊丽 高必达 |
| 作者单位： | 1. 浙江理工大学生物工程研究所,中国,杭州,310018;湖南农业大学生物安全科技学院,中国,长沙,410128;大连民族学院生命科学学院,中国,大连,116600 2. 浙江理工大学生物工程研究所,中国,杭州,310018 3. 湖南农业大学生物安全科技学院,中国,长沙,410128 |
| 基金项目： | 国家“863”资助项目(2002AA24112A) |
| 摘 要： | 利用实时荧光PCR方法,以18srRNA为内参照,对半夏植株及相应组培苗携带的黄瓜花叶病毒(Cu-cumber Mosaic Virus,CMV)含量进行了相对定量检测.并以DAS-ELISA和RT-PCR方法检测结果相比较.结果表明,相比原始植株,组培苗的病毒含量大大降低.以带病毒半夏母株为外植体培养的组培苗,其病毒含量以叶片中的相对较高,在块茎和愈伤组织中病毒相对较低.而ELISA方法在检测病毒含量低的样品时有假阴性结果;RT-PCR方法灵敏度可靠但不适宜精确量化. |
| 关 键 词： | 实时荧光PCR 黄瓜花叶病毒 反转录-PCR 双夹心酶联免疫反应 半夏 |
| 文章编号： | 1000-2537（2006）03-0103-04 |
| 收稿时间： | 2006-04-06 |
| 修稿时间： | 2006年4月6日 |
On Detection of Pinellia Ternata CMV by Real-time PCR with 18srDNA as endogenous Reference |
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| HE Yubo,CHEN Ji-shuang,CHEN Hai-min,FENG Jun-li,GAO Bi-da.On Detection of Pinellia Ternata CMV by Real-time PCR with 18srDNA as endogenous Reference[J].Journal of Natural Science of Hunan Normal University,2006,29(3):103-106. | |
| Authors: | HE Yubo CHEN Ji-shuang CHEN Hai-min FENG Jun-li GAO Bi-da |
| Institution: | 1. Insitute of Biotechnology, Zhejiang Institute of Science and Technology, Hangzhou 310018, China; 2. College of bio-safety science and technology, Htman Agricultural University, Changsha 410128, China; 3. College of Life Science, Dalian Nationalities University, Dalian 116600, China |
| Abstract: | With 18srDNA as as endogenous reference,relative quantification detection of Cucumber Mosaic Virus(CMV) in Pinellia Ternata and its tissue cultured plantlets was carried by using Real-time PCR,and combined with methods of DAS-ELISA and RT-PCR.The results showed that CMV concentration in tissue cultured plantlets was decreased greatly compared with the parent plantlet.CMV quantity in leaf of tissue cultured plantlets which come from explant of CMV positive Pinellia Ternata was relatively higher than that of tubercles and callus.There were fake positive results while using ELISA method to detect samples of low concentration virus.RT-PCR method is sensitive but not quantified exactly to detected samples. |
| Keywords: | Real-time PCR CMV RT-PCR DAS-ELISA Pinellia Ternata |
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