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.
PCR-based assay for the detection of Curtobacterium flaccumfaciens pv. flaccumfaciens in bean seeds / S. TEGLI; A. SERENI; G. SURICO. - In: LETTERS IN APPLIED MICROBIOLOGY. - ISSN 0266-8254. - STAMPA. - 35:(2002), pp. 331-337. [10.1046/j.1472-765X.2002.01187.x]
PCR-based assay for the detection of Curtobacterium flaccumfaciens pv. flaccumfaciens in bean seeds.
TEGLI, STEFANIA;SURICO, GIUSEPPE
2002
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.File | Dimensione | Formato | |
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