利用粗汁液提取物进行用于植原体检测的逆转录聚合酶链反应。

Reverse transcription-PCR for phytoplasma detection utilizing crude sap extractions.

作者信息

Margaria Paolo, Palmano Sabrina

机构信息

Istituto di Virologia Vegetale, CNR, Torino, Italy.

出版信息

Methods Mol Biol. 2013;938:283-9. doi: 10.1007/978-1-62703-089-2_24.

DOI:10.1007/978-1-62703-089-2_24

PMID:22987424

Abstract

Phytoplasmas are routinely detected by nucleic acid-based techniques. These approaches rely on enriched phytoplasma DNA extracts of good quality, following labor intensive and time-consuming purification protocols. Here we describe a very rapid, specific, sensitive, and reliable method for flavescence dorée phytoplasma detection, based on real-time Taqman(®) reverse transcription-PCR of the 16S rRNA. The protocol is particularly useful for large-scale screening of vineyards and nurseries, pathogen surveys, and field epidemiological studies.

摘要

植原体通常通过基于核酸的技术进行检测。这些方法依赖于经过劳动强度大且耗时的纯化方案后获得的高质量富集植原体DNA提取物。在此，我们描述了一种基于16S rRNA的实时Taqman®逆转录PCR的、用于检测葡萄黄化植原体的非常快速、特异、灵敏且可靠的方法。该方案对于葡萄园和苗圃的大规模筛查、病原体调查以及田间流行病学研究特别有用。