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目的调查临床分离45株多药耐药不动杆菌属基因同源性,探讨产β-内酰胺酶基因型在多药耐药不动杆菌属中所起的作用。方法采用纸片扩散法筛选多药耐药不动杆菌属;WHONET5.4分析耐药趋势变化;改良三维试验检测β-内酰胺酶表型;PFGE不动杆菌属同源性;聚合酶链反应检测耐β-内酰胺酶基因型对扩增阳性产物进行测序,分析β-内酰胺酶类型突变位点。结果45株不动杆菌属中3株(7.32%)未检出β-内酰胺酶基因型,28株产OXA-23型(68.3%)、10株产I MP型(24.4%)、13株产TEM型(31.7%)、18株产CTX-M型(43.9%)、6株产PER型(14.6%)、7株产AmpC型β-内酰胺酶(17.1%),未检出产SHV型β-内酰胺酶;24株(58.5%)同时产生≥2种酶型;对10号菌OXA扩增产物进行测序证实为OXA-23型,与genebank公布的OXA-23型基因100%同源。结论不动杆菌属可产生多种β-内酰胺酶,是造成多药耐药的重要机制之一。
Objective To investigate the clinical isolates of 45 multidrug-resistant Acinetobacter spp. Genes and investigate the role of β-lactamase genotype in multidrug-resistant Acinetobacter. Methods Multidrug-resistant Acinetobacter was screened by disk diffusion method. WHONET5.4 was used to analyze the trend of drug resistance. The modified three-dimensional test was used to detect the β-lactamase phenotype. The genus PFGE consisted of Acinetobacter species. Polymerase chain reaction The beta-lactamase-resistant genotypes were tested for amplification-positive products and the beta-lactamase-type mutation sites were analyzed. Results No β-lactamase genotypes were detected in 3 strains of Acinetobacter (7.32%), 28 strains were OXA-23 type (68.3%), 10 strains were MP type (24.4%) and 13 strains 18 strains of CTX-M (43.9%), 6 strains of PER (14.6%) and 7 strains of AmpC β-lactamase (17.1%) were produced by TEM, 31.7% 24 strains (58.5%) produced more than 2 types of enzyme at the same time. The OXA amplification product of strain 10 was confirmed as OXA-23 type, which was 100% homologous with the gene type OXA-23 source. Conclusion Acinetobacter can produce a variety of β-lactamase, which is one of the important mechanisms that cause multi-drug resistance.