Record nummer 2070639
Titel Validation report of a real-time PCR assay for the identification of Clavibacter michiganensis subsp. michiganensis (Cmm) isolates
toon extra info.
Adriaan Vermunt and Floor den Otter
Auteur(s) Vermunt, A. ; Otter, F. den
Uitgever [Delfgauw] : Groen Agro Control
Jaar van uitgave 2010
Pagina's 1 online resource (PDF, 14 pages) illustrations
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Trefwoorden (cab) clavibacter michiganensis subsp. michiganensis / real time pcr / polymerase-kettingreactie / tomaten / solanum lycopersicum / plantenziektebestrijding / moleculaire detectie / plantenziekten
Rubrieken Landbouwkundige bacteriologie (algemeen) / Biologische technieken
Publicatie type Boek
Taal Engels
Toelichting (Engels) This reports presents complete validation data related to recently developed primer-probe combinations for the identification of Clavibacter michiganensis subsp. michiganensis (Cmm) isolates with the use of a real-time PCR assay. The primers and probes were developed on two loci: on the intergenic spacer between the 16S and 23S ribosomal RNA genes (IGS) and on the pathogenity-1 gene (Pat-1), which is located on the plasmid pCM2. The validated method is qualitative and consists of a DNA extraction from pure colony material followed by a duplex real-time PCR assay based on the primer-probe combinations for IGS and Pat-1. The method is very specific, because only the 20 tested pathogenic Cmm isolates were positive for both primer-probe combinations. Negative results were obtained from 12 Cmm look-alikes, 4 Cm isolates and 9 other tomato pathogens. Tests on repeatability, reproducibility and robustness of this assay performed well.
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