通过逆转录聚合酶链反应（RT-PCR）和免疫捕获逆转录聚合酶链反应在番杏和甜菜中检测甜菜黄化病毒

Detection of beet yellows virus by RT-PCR and immunocapture RT-PCR in Tetragonia expansa and Beta vulgaris.

作者信息

Kundu K, Rysánek P

机构信息

Department of Plant Protection, Czech University of Agriculture, 165 21 Prague 6, Czech Republic.

出版信息

Acta Virol. 2004;48(3):177-82.

PMID:15595212

Abstract

Two sensitive methods, RT-PCR with phenol-extracted RNA or Triton X-100-released RNA and immunocapture RT-PCR (IR-RT-PCR) were used for the detection of Beet yellows virus (BYV) in young and old leaves of Tetragonia expansa and sugar beet (Beta vulgaris) and in sugar beet roots. Four oligonucleotide primer pairs proved suitable for the detection of BYV. The release of BYV RNA with Triton X-100 was shown to be a very effective and easy as compared to isolation of total RNA by phenol extraction with the same or higher sensitivity of subsequent PCR. Using the Triton X-100 release of RNA and IC-RT-PCR the sensitivity of detection was so high that pg amounts of BYV RNA occurring in dilutions up to 10(-6) of saps from young Tetragonia and sugar beet leaves could be detected.

摘要

两种灵敏的方法，即使用苯酚提取的RNA或经Triton X-100释放的RNA进行逆转录聚合酶链式反应（RT-PCR）以及免疫捕获RT-PCR（IR-RT-PCR），用于检测番杏和甜菜（Beta vulgaris）的幼叶与老叶以及甜菜根中的甜菜黄化病毒（BYV）。四对寡核苷酸引物被证明适用于BYV的检测。与通过苯酚提取总RNA相比，用Triton X-100释放BYV RNA被证明是一种非常有效且简便的方法，后续PCR的灵敏度相同或更高。使用Triton X-100释放RNA和免疫捕获RT-PCR，检测灵敏度极高，以至于可以检测到番杏和甜菜幼叶汁液稀释至10(-6)时所含的皮克量的BYV RNA。