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目的检测临床分离的72株多药耐药铜绿假单胞菌(PAE)整合子分布情况,并对其中一株vim2基因阳性菌株整合子结构及定位进行分析。方法通过药敏试验检测铜绿假单胞菌的MIC,以PCR扩增多药耐药PAE的vim2耐药基因和Ⅰ类整合子,电泳胶回收DNA片段并测序,将测序结果以Pubmed在线软件BLAST进行序列比对分析。结果 72株非重复多药耐药铜绿假单胞菌的intⅠ基因阳性率高达99.0%,在一株多药耐药铜绿假单胞菌(命名为PAE vim2)中检测到双整合酶,两整合酶及其相连的不同基因盒均位于细菌染色体上,其中一个结构为intⅠ-sul1,位于Tn21样转座子中,另一个为Tn402样的Ⅰ类整合子,后者携带vim2等基因盒。结论多药耐药PAE整合子的携带率高,且在一株多药耐药铜绿假单胞菌中发现双整合酶耐药结构,可能是引起耐药基因水平转移的遗传物质基础。
Objective To detect the distribution of 72 clinical isolates of multidrug-resistant Pseudomonas aeruginosa (PAE), and to analyze the integron structure and localization of one of the vim2-positive strains. Methods MIC of Pseudomonas aeruginosa was detected by drug susceptibility test. The vim2 resistance gene and class Ⅰ integrons of multidrug-resistant PAE were amplified by PCR. The DNA fragments were recovered and sequenced. The sequencing results were based on Pubmed online software BLAST Sequence alignment analysis. Results The positive rate of intⅠ gene in 72 non-repetitive multidrug-resistant Pseudomonas aeruginosa strains was as high as 99.0%. Double integrase was detected in a multidrug-resistant Pseudomonas aeruginosa named PAE vim2. The enzyme and its associated gene cassettes are located on bacterial chromosomes, one of them being intI-sul1, located in Tn21-like transposon and the other being Tn402-like class I integron, which carries the vim2 gene cassette. Conclusions The carrier rate of multidrug-resistant PAE integron is high, and the drug-resistant structure of double integrase is found in a multidrug-resistant Pseudomonas aeruginosa, which may be the genetic material basis for the horizontal metastasis of drug-resistant genes.