Comparison of specificity and sensitivity of immunochemical and molecular techniques for determination of Clavibacter michiganensis subsp. michiganensis

0346909 - BC 2011 RIV CZ eng J - Journal Article
Kokošková, B. - Mráz, Ivan - Fousek, Jan
Comparison of specificity and sensitivity of immunochemical and molecular techniques for determination of Clavibacter michiganensis subsp. michiganensis.
Folia Microbiologica. Roč. 55, č. 3 (2010), s. 239-244. ISSN 0015-5632. E-ISSN 1874-9356
R&D Projects: GA MZe QH71229
Institutional research plan: CEZ:AV0Z50510513
Keywords : Clavibacter michiganensis subsp. michiganensis (Cmm) * PCR * ELISA * IFAS
Subject RIV: EF - Botanics
Impact factor: 0.977, year: 2010

Detection of Cmm, causing bacterial canker of tomato, was verified using PTA-ELISA, IFAS and with published and own PCR primers. The specificity of this technique was determined with 15 plant-pathogenic and 4 common, saprophytic bacteria. With IFAS, crossreactions were found for Pantoea dispersa, P. agglomerans and Rahnella aquatilis, and with PTA-ELISA for Curtobacterium flaccumfaciens, Pectobacterium atrosepticum and Dickeya sp. Cross-reactions with subspecies other than michiganensis were also found using both methods. Molecular methods were optimized by verification of annealing temperatures and times for both primers. After this optimization, both primer pairs produced positive reaction only with Cmm. By means of PTA-ELISA and IFAS, Cmm strains were detected at a concentration up to 105 CFU/mL and 103 CFU/mL, respectively. The PCR test with bacterial cell suspensions reached a sensitivity of 103 CFU/mL with our designed primers and 104 CFU/mL with Dreier’s primer pair.
Permanent Link: http://hdl.handle.net/11104/0187812