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目的了解整合子在广州地区铜绿假单胞菌中的分布状况及其主要类型,研究I类整合子的结构特征并探讨其与细菌多重耐药之间的相关性。方法采用PCR检测3类整合子整合酶的编码基因,并扩增整合子的可变区,运用DNA测序技术分析整合子可变区的耐药基因及其排列顺序,利用生物信息学的方法对检测到的可变区基因序列进行比对分析。结果在20株产金属-内酰胺酶铜绿假单胞菌中,15株检测到I类整合子,未检测到II类和III类整合子,其中一株铜绿假单胞菌整合子全长序列含有I类整合子整合酶、AacA4氨基糖甙乙酰转移酶、IMP-25金属-内酰胺酶、OXA-30-内酰胺酶和CatB3氯霉素乙酰转移酶的编码基因及其相应的基因重组位点,GenBank登录号为EU588392,其中IMP-25为一个新的金属-内酰胺酶亚型。结论广州地区铜绿假单胞菌中存在的整合子类型是I类整合子,并且该整合子可变区携带多种耐药基因盒,其中一个是金属-内酰胺酶新亚型IMP-25,在分子水平上阐述了I类整合子参与铜绿假单胞菌对抗生素的耐药性和多重耐药性。
Objective To investigate the distribution and main types of integron in Pseudomonas aeruginosa in Guangzhou and to study the structural characteristics of the class I integron and to explore its correlation with bacterial multi-drug resistance. Methods Three kinds of integrase integrase genes were amplified by polymerase chain reaction (PCR), and the variable regions of the integron were amplified. The resistance genes of the integron variable regions were analyzed by DNA sequencing. The bioinformatics methods Detected variable region gene sequences for alignment analysis. Results Of the 20 strains of L-Pseudomonas aeruginosa, 15 strains of type I integrons were detected and no class II or III integrons were detected, of which one full-length sequence of Pseudomonas aeruginosa integron The genes encoding class I integron integrase, AacA4 aminoglycoside acetyltransferase, IMP-25 metal-lactamase, OXA-30-lactamase and CatB3 chloramphenicol acetyltransferase and their corresponding gene recombination sites Point, GenBank accession number EU588392, in which IMP-25 is a new metal-lactamase subtype. CONCLUSIONS: The integron type present in Pseudomonas aeruginosa in Guangzhou is a type I integron, and the integron variable region carries multiple drug resistance gene cassettes, one of which is a new type of metal-lactamase IMP-25, At the molecular level, the class I integrins are involved in the resistance and multi-drug resistance of antibiotics to Pseudomonas aeruginosa.