当前位置: 首页 > 文章 > 苹果褪绿叶斑病毒一步法RT-PCR检测 西北农业学报 2006,15 (4) 67-69
Position: Home > Articles > A Study on the One-step RT-PCR for Detection of Apple Chlorotic Leaf Spot Virus Acta Agriculturae Boreali-occidentalis Sinica 2006,15 (4) 67-69

苹果褪绿叶斑病毒一步法RT-PCR检测

作  者:
张强;牛建新;李西萍
单  位:
石河子大学农学院园艺系
关键词:
一步法RT-PCR;检测;苹果褪绿叶斑病毒(ACLSV)
摘  要:
RT-PCR技术在植物病毒检测方面有着快速、灵敏、特异性强等优点。然而传统的RT-PCR是在两个酶(逆转录酶和Taq酶)分别催化下在两个体系中的两步反应,即由RNA逆转录为cDNA,再以模板cDNA进行PCR扩增,操作步骤比较繁琐,而且很容易造成交叉污染。笔者建立的一步法RT-PCR是在逆转录酶(M-MLV)和Taq DNA聚合酶共同作用下,只需在一个反应管中就可以完成反转录和cDNA扩增的全过程,两个步骤一步完成,以达到对苹果褪绿叶斑病毒快速、简便、敏感的检测。
译  名:
A Study on the One-step RT-PCR for Detection of Apple Chlorotic Leaf Spot Virus
作  者:
ZHANG Qiang,NIU Jian-xin~* and LI Xi-ping(Horticulture Department of Agricultural College,Sihezi University,Shihezi Xinjiang 832003,China)
关键词:
One-step reverse-transcription polymerase chain reaction(RT-PCR) method;Detecting;Apple chlorotic leaf spot virus
摘  要:
RT-PCR method has many advantages such as rapid,sensitive,special at the detection of plant virus.However,traditional RT-PCR method includes two-reactions,which are catalyzed by two enzymes―Reverse Transcriptase and Taq DNA polymerease respectively.The reactions are: RNA is template to synthesize the cDNA,then the cDNA is template to complete the PCR amplification.The process is so complex that is easy to be contaminated and be found false positive.The assay established a one-step RT-PCR method,that is,under the co-effection of Reverse Transcriptase(M-MLV) and Taq DNA polymerase,the reverse transcription and the amplification can be completed in one tube.The two process were accomplished by one step,which reach to detect Apple chlorotic leaf spot virus rapidly,simply and sensitively.

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