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Comparison of specificity and sensitivity of immunochemical and molecular techniques for determination of Clavibacter michiganensis subsp. michiganensis
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SYSNO ASEP 0346909 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Comparison of specificity and sensitivity of immunochemical and molecular techniques for determination of Clavibacter michiganensis subsp. michiganensis Author(s) Kokošková, B. (CZ)
Mráz, Ivan (BC-A)
Fousek, Jan (BC-A)Number of authors 3 Source Title Folia Microbiologica. - : Springer - ISSN 0015-5632
Roč. 55, č. 3 (2010), s. 239-244Number of pages 6 s. Language eng - English Country CZ - Czech Republic Keywords Clavibacter michiganensis subsp. michiganensis (Cmm) ; PCR ; ELISA ; IFAS Subject RIV EF - Botanics R&D Projects QH71229 GA MZe - Ministry of Agriculture (MZe) CEZ AV0Z50510513 - UMBR-M, BC-A (2005-2011) UT WOS 000278469400007 DOI 10.1007/s12223-010-0035-4 Annotation 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. Workplace Biology Centre (since 2006) Contact Dana Hypšová, eje@eje.cz, Tel.: 387 775 214 Year of Publishing 2011
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