使用[32P]DNA探针的斑点印迹法用于灵敏检测植物中的鳄梨日斑病和其他类病毒。

Dot-blot procedure with [32P]DNA probes for the sensitive detection of avocado sunblotch and other viroids in plants.

作者信息

Barker J M, McInnes J L, Murphy P J, Symons R H

出版信息

J Virol Methods. 1985 Feb;10(2):87-98. doi: 10.1016/0166-0934(85)90094-1.

Abstract

Avocado sunblotch viroid (ASBV) has been detected down to a level of about 20 pg per gram fresh weight of leaves by the use of a dot-blot hybridization procedure and partially purified nucleic acid extracts. Three [32P]DNA probes were compared, two prepared from full-length ASBV clones in the single-strand M13mp93 vector and the other by primer extension on purified ASBV. All three probes gave the same sensitivity of detection of ASBV. The methods developed have also been used successfully for the routine detection of potato spindle tuber viroid, citrus exocortis viroid, and coconut cadang cadang viroid.

摘要

利用斑点杂交程序和部分纯化的核酸提取物，已检测到鳄梨日斑类病毒（ASBV）在叶片鲜重每克约20皮克的水平。比较了三种[32P]DNA探针，两种由单链M13mp93载体中的全长ASBV克隆制备，另一种通过对纯化的ASBV进行引物延伸制备。所有三种探针检测ASBV的灵敏度相同。所开发的方法也已成功用于马铃薯纺锤块茎类病毒、柑橘裂皮类病毒和椰子死亡类病毒的常规检测。