| 检测砂梨潜隐病毒的IC-RT-PCR和TC-RT-PCR的研究 |
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| 引用本文： | 邓晓云,洪霓,胡红菊,王国平.检测砂梨潜隐病毒的IC-RT-PCR和TC-RT-PCR的研究[J].果树学报,2004,21(6):569-572. |
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| 作者姓名： | 邓晓云 洪霓 胡红菊 王国平 |
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| 作者单位： | 1. 华中农业大学植物科技学院,湖北,武汉,430070
2. 湖北省农业科学院果茶蚕桑研究所,湖北,武汉,430209 |
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| 基金项目： | 科技部863课题(项目编号:2001AA241142)。 |
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| 摘 要： | 以砂梨为试材,在生物学和血清学检测的基础上,采用免疫捕作RT-PCR穴IC-RT-PCR雪和试管捕作RT-PCR穴TC-RT-PCR雪技术,对苹果褪绿叶斑病毒(Applechloroticleafspotvirus,ACLSV)和苹果茎沟病毒(Applestemgroovingvirus,ASGV)进行了检测分析。结果表明,TC/IC-RT-PCR均能有效检测梨粗提液中的ACLSV和ASGV,分别获得了大小约358bp和499bp的目标扩增片段;但IC-RT-PCR检测ACLSV的效果受到病毒分离株间血清学关系影响。与传统的RT-PCR相比,IC/TC-RT-PCR不仅灵敏度更高,而且不需提取总RNA,可以简化操作程序和减少苯酚、氯仿类有机试剂对人体的伤害和对环境的污染,适合于大量梨样品的病毒快速灵敏检测。
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| 关 键 词： | 砂梨 ACLSV ASGV IC-RT-PCR TC-RT-PCR |
| 文章编号： | 1009-9980(2004)06-569-04 |
| 修稿时间： | 2004年3月31日 |
Detection of Latent Viruses in Pyrus pyrifolia by IC- RT-PCR and TC-RT-PCR |
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| Deng Xiaoyun,Hong Ni,Hu Hongju,Wang Guoping.Detection of Latent Viruses in Pyrus pyrifolia by IC- RT-PCR and TC-RT-PCR[J].Journal of Fruit Science,2004,21(6):569-572. |
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| Authors: | Deng Xiaoyun Hong Ni Hu Hongju Wang Guoping |
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| Institution: | Deng Xiaoyun1,Hong Ni1,Hu Hongju2,and Wang Guoping1* |
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| Abstract: | In this study, immunocapture (IC)-RT-PCR and tube capture(TC) RT-PCR were applied for detecting two major viruses Apple stem grooving virus (ASGV) and Apple chlorotic leaf spot virus (ACLSV) in Pyrus pyrifolia samples. Results showed that both TC-RT-PCR and IC-RT-PCR could be successfully used to detect ASGV and ACLSV in pear crude extracts and their expected fragments about 358 bp and 499 bp were amplified. But the results for the detection of ACLSV by IC-RT-PCR depended on the serological relation distance between virus isolates. Compared with usual RT-PCR, IC/TC-RT-PCR not only have a higher sensitivity, but also can simplify the operation procedure and decrease the effect of phenol and chloroform on researchers and environment. IC/TC-RT-PCR is suitable for rapid and high sensitive detection of pear viruses in large amount samples. |
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| Keywords: | Pyrus pyrifolia Latent viruses IC-RT-PCR TC-RT-PCR |
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