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基于聚合酶链式反应的菜豆萎蔫病菌检测方法在菜豆种子中的应用。 (注:Curtobacterium flaccumfaciens pv. flaccumfaciens为菜豆萎蔫病菌,pv. 表示致病变种,这里翻译时按专业术语习惯调整了表述使更通顺自然)

PCR-based assay for the detection of Curtobacterium flaccumfaciens pv. flaccumfaciens in bean seeds.

作者信息

Tegli S, Sereni A, Surico G

机构信息

Dipartimento di Biotecnologie Agrarie (DiBA) - Sezione di Patologia Vegetale, Università degli Studi di Firenze, Firenze, Italy.

出版信息

Lett Appl Microbiol. 2002;35(4):331-7. doi: 10.1046/j.1472-765x.2002.01187.x.

Abstract

AIMS

To develop a PCR-based protocol for the rapid, sensitive and specific detection of Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff) in bean seeds.

METHODS AND RESULTS

A pair of PCR primers (CffFOR2-CffREV4), targeting the sequence of a cloned DNA fragment of 550 bp amplified in Repetitive-sequence-based-PCR (Rep-PCR) experiments, were designed and shown to specifically amplify a 306-bp DNA fragment using Cff DNA as template. Moreover, this PCR protocol was demonstrated to successfully detect Cff in naturally infected bean seeds in 36 h.

CONCLUSIONS

A specific, highly sensitive and rapid PCR assay for the detection of Cff was achieved.

SIGNIFICANCE AND IMPACT OF THE STUDY

Cff is a seed-borne bacterium on the EPPO A2 quarantine list; this procedure may be useful for routine diagnosis of Cff, overcoming the problems of conventional techniques.

摘要

目的

开发一种基于聚合酶链反应(PCR)的方法,用于快速、灵敏且特异性地检测菜豆种子中的萎蔫短小杆菌萎蔫致病变种(Cff)。

方法与结果

设计了一对PCR引物(CffFOR2 - CffREV4),其靶向基于重复序列PCR(Rep - PCR)实验中扩增出的550 bp克隆DNA片段序列,并证明以Cff DNA为模板时能特异性扩增出306 bp的DNA片段。此外,该PCR方法被证明能在36小时内成功检测出自然感染的菜豆种子中的Cff。

结论

实现了一种用于检测Cff的特异性强、灵敏度高且快速的PCR检测方法。

研究的意义与影响

Cff是欧洲和地中海植物保护组织(EPPO)A2检疫名单上的一种种传细菌;该方法可能有助于Cff的常规诊断,克服传统技术存在的问题。

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