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


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.


CONCLUSIONS
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.
D000192:Actinomycetales / Q000235:genetics / Q000302:isolation & purification*; D003001:Cloning, Molecular; D017931:DNA Primers; D007887:Fabaceae / Q000382:microbiology; D005516:Food Microbiology; D016908:Gram-Positive Bacterial Infections / Q000382:microbiology; D010935:Plant Diseases / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012091:Repetitive Sequences, Nucleic Acid; D012639:Seeds / Q000382:microbiology*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
D017931:DNA Primers