Tegli S, Sereni A, Surico G
Dipartimento di Biotecnologie Agrarie (DiBA) - Sezione di Patologia Vegetale, Università degli Studi di Firenze, Firenze, Italy.
Lett Appl Microbiol. 2002;35(4):331-7. doi: 10.1046/j.1472-765x.2002.01187.x.
To develop a PCR-based protocol for the rapid, sensitive and specific detection of Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff) in bean seeds.
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.
A specific, highly sensitive and rapid PCR assay for the detection of Cff was achieved.
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.
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）。
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。
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。
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。
