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目的了解大连地区分离的耐碳青霉烯类铜绿假单胞菌的金属β-内酰胺酶、整合子I和ISCR1的分布情况,并分析其基因多态性特征。方法收集临床分离的89株耐亚胺培南铜绿假单胞菌,PCR检测金属酶、整合子I、ISCR1耐药基因。脉冲场凝胶电泳(PFGE)进行细菌基因分型。结果 89株亚胺培南耐药的铜绿假单胞菌中,ISCR1基因阳性菌25株(25/89,28%),其中84%(21/25)为多重耐药,5类及以上药物耐药的菌株占64%(16/25),金属酶基因阳性为11株(11/89,12%),其中有8株携带IMP-1基因,3株携带VIM-2,整合子I基因阳性43株(43/89,48%)。但携带金属酶的菌株整合子I、ISCR1基因扩增均为阴性;PFGE分型结果显示:89株耐亚胺培南的铜绿假单胞菌分为15个基因型(A~O),其中A型46株、B型16株、C型4株、D型5株、E型4株、F型3株、G型2株、H型2株,I型~O型各有1株。基因型集中的A型~G型,各型中的菌株来源于不同医院,呈多态性,每群均存在克隆株。结论基因捕获元件整合子I基因及ISCR1广泛分布在大连地区耐碳青霉烯类铜绿假单胞菌中,并与细菌的多重耐药、泛耐药显著相关,特别是ISCR1基因;大连地区整合子I、ISCR1并未携带金属酶基因盒。PFGE结果提示本地区铜绿假单胞菌具有基因多态性,但仍存在高度同源性的流行优势基因型。
Objective To investigate the distribution of metal β-lactamase, integron I and ISCR1 in carbapenem-resistant Pseudomonas aeruginosa isolated in Dalian, and to analyze the gene polymorphism. Methods 89 strains of imipenem - resistant Pseudomonas aeruginosa isolated from clinical isolates were collected. The metalloenzymes, integron I and ISCR1 resistance genes were detected by PCR. Pulsed-field gel electrophoresis (PFGE) was used to genotype the bacteria. Results Among 89 strains of imipenem-resistant Pseudomonas aeruginosa, 25 (25/89, 28%) were positive for ISCR1 gene, of which 84% (21/25) were multi-drug resistant and 5 or more The resistant strains accounted for 64% (16/25), and 11 (89.8%) were positive for the metalloenzyme gene. Among them, 8 strains carried IMP-1 gene and 3 strains carried VIM-2 and integron I gene Positive 43 (43 / 89,48%). However, the genotypes of integron I and ISCR1 carrying metalloenzyme were all negative. The results of PFGE typing showed that 89 strains of imipenem-resistant Pseudomonas aeruginosa were divided into 15 genotypes (A ~ O) 46 strains of type A, 16 strains of type B, 4 strains of type C, 5 strains of type D, 4 strains of type E, 3 strains of type F, 2 strains of type G, 2 strains of type H and 1 strain of type I to 0 respectively. The genotypes concentrated in type A to type G, each type of strains from different hospitals, was polymorphic, clonal strains exist in each group. Conclusion The gene capture element integron I gene and ISCR1 are widely distributed in carbapenem-resistant Pseudomonas aeruginosa in Dalian, and are significantly associated with multi-drug resistance and pan-drug resistance of bacteria, especially the ISCR1 gene; Subset I, ISCR1 does not carry the metalloenzyme gene cassette. PFGE results suggest that the region Pseudomonas aeruginosa has genetic polymorphism, but there is still a high degree of homology prevalence of genotypes.