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| Real-time Fluorescent Quantitative PCR Detection for Burkholderia gladioli pv. alliicola |
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Abstract Burkholderia gladioli pv. alliicola is an important plant pathogen. This research was to establish the specific, sensitive and fast Taqman Real-time PCR for detection of B. gladioli pv. alliicola. The specific genome region of B. gladioli pv. alliicola was used for the design of Taqman probe and specific primers. The reaction conditions was optimized. Fourteen reference strains were used for the specificity test, the sensitivity of the Real-time PCR was tested as well. The sensitivity of bacterial suspension of Real-time PCR reached 1.02×102 CFU/mL and the sensitivity of DNA reached 1.73×10-4 ng/μL. Total reaction time took about 1 h or so. Among 14 reference strains belong to the genus of Burkholderia, Real-time PCR showed an extremely high specificity. The simulation test was performed on the onion (Allium cepa) seeds mixed with bacterial suspension of B. gladioli pv. alliicola, the results revealed that the Real-time PCR was practical in the test of B. gladioli pv. alliicola. The Real-time PCR established in this research is useful for the diagnostics of B. gladioli pv. alliicola. It provides an easy, fast and accurate method for molecular identification of B. gladioli pv. alliicola.
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Received: 12 July 2012
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