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多重逆转录聚合酶链反应,一种用于同时检测引起水稻东格鲁病的DNA和RNA病毒的新技术。

Multiplex RT-PCR, a novel technique for the simultaneous detection of the DNA and RNA viruses causing rice tungro disease.

作者信息

Periasamy M, Niazi F R, Malathi V G

机构信息

Plant Virology Unit, Division of Plant Pathology, Indian Agricultural Research Institute, New Delhi 110012, India.

出版信息

J Virol Methods. 2006 Jun;134(1-2):230-6. doi: 10.1016/j.jviromet.2006.01.010. Epub 2006 Feb 21.

Abstract

Rice tungro, economically the most important viral disease of rice, is a complex disease caused by two morphologically and genomically dissimilar viruses, Rice tungro bacilliform virus (RTBV), a double stranded DNA virus replicating through RNA intermediate and Rice tungro spherical virus (RTSV), a single stranded RNA virus with 3' poly (A) tail. A novel multiplex RT-PCR technique for the simultaneous detection of RTBV and RTSV from the total RNA extracted from tungro-infected plants has been developed. It involves a one-step reaction initiating first strand cDNA synthesis by oligo (dT) primer with poly (A) tailed RTBV transcript and RTSV genomic RNA as template for the PCR amplification. The results indicate that adaptation of this technique will strengthen the screening for tungro resistance among rice varieties and hybrids.

摘要

水稻东格鲁病是水稻在经济上最重要的病毒病,是由两种形态和基因组不同的病毒引起的复合病害,即水稻东格鲁杆状病毒(RTBV),一种通过RNA中间体复制的双链DNA病毒,以及水稻东格鲁球状病毒(RTSV),一种具有3'聚(A)尾的单链RNA病毒。已经开发出一种新颖的多重RT-PCR技术,用于从感染东格鲁病的植物中提取的总RNA中同时检测RTBV和RTSV。它涉及一步反应,以带有聚(A)尾的RTBV转录本和RTSV基因组RNA为模板,通过寡聚(dT)引物启动第一链cDNA合成,用于PCR扩增。结果表明,采用该技术将加强对水稻品种和杂交种中东格鲁病抗性的筛选。

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