| 英文摘要 |
"A specific PCR (polymerase chain reaction) primer pair has been developed to detect Acidovorax anthurii using RAPD (random amplified polymorphic DNA). A 1813 bp DNA fragment was amplified from genomic DNA of A. anthurii using OPW-2 primer, after screening one hundred and sixty random primers in RAPD. The amplified DNA was purified, cloned into a pCR II-TOPO vector, and sequenced. A primer pair Ant 2F/Ant 2R based on 1813-bp DNA sequence was designed and tested for specificity. The primer pair could amplify a DNA fragment of 343 bp that was specific to Acidovorax anthurii. No DNA fragment was amplified by the same primer pair from other 21 bacterial species in 8 genera, including non-target pathogens and saprophytes. The designed primer pair Ant 2F/Ant 2R was highly sensitive and could amplify the 343-bp fragment from A. anthurii DNA between 10-50 pg obtained from 4.5×101 cfu/ml bacterial cells. Non-target bacterial DNA did not affect the efficiency of specific amplification of A. anthurii in PCR assays with the primer pair Ant 2F/Ant 2R. Successful identification of A. anthurii could be achieved within 3-4 hrs using PCR with specific primer set. Based on the data provided, we conclude that the primer pair Ant 2F/Ant 2R will be a useful tool for rapid identification and diagnosis of Acidovorax anthurii." |
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