Roy B P, AbouHaidar M G, Alexander A
Department of Botany, University of Toronto, Ontario, Canada.
J Virol Methods. 1989 Feb;23(2):149-55. doi: 10.1016/0166-0934(89)90128-6.
Diseases caused by potato spindle tuber viroid (PSTV) are of significant agronomic importance, and early detection is vital. The objective of this paper was to synthesize biotinylated RNA probes in order to develop a specific, sensitive, and reliable assay system for detection of PSTV in infected plants. RNA probes were prepared by in vitro transcription of cloned PSTV, and were labelled with either biotin-11-UTP or [32P]UTP. Partially purified total RNAs from healthy and from PSTV-infected plants were spotted onto nitrocellulose filters and hybridized with either biotin-labelled or 32P-labelled probes. Our results showed that the sensitivity of biotinylated probes was similar to that for the 32P-labelled probes.
由马铃薯纺锤块茎类病毒（PSTV）引起的病害具有重要的农学意义，早期检测至关重要。本文的目的是合成生物素化RNA探针，以便开发一种用于检测受感染植物中PSTV的特异性、灵敏且可靠的检测系统。通过对克隆的PSTV进行体外转录制备RNA探针，并用生物素-11-UTP或[32P]UTP进行标记。将来自健康和PSTV感染植物的部分纯化总RNA点样到硝酸纤维素滤膜上，并用生物素标记或32P标记的探针进行杂交。我们的结果表明，生物素化探针的灵敏度与32P标记探针的灵敏度相似。
