摘要
      严重耐多药细菌引起的感染急剧增加是全球健康问题。在这项研究中，我们对从一名意大利患者的手术伤口和直肠拭子中分离出的肺炎克雷伯菌、奇异芽孢杆菌、阴沟肠杆菌和鲍曼不动杆菌的抗微生物耐药性基因（ARGs）进行了表征。细菌鉴定采用MALDI-TOF MS进行，抗菌药物敏感性采用Vitek 2系统进行。使用下一代测序（NGS）方法（MiSeq Illumina仪器）对ARGs进行表征。肺炎克雷伯菌、奇异芽孢杆菌和阴沟肠杆菌对大多数β-内酰胺类和β-内胺/β-内酶抑制剂组合具有耐药性。鲍曼不动杆菌菌株仅对粘菌素敏感。在所有肠杆菌中都检测到质粒（IncN、IncR、IncFIB、ColRNAI和Col（MGD2））的存在，但在鲍曼不动杆菌菌株中没有检测到。IncN质粒和blaNDM-1基因在肺炎克雷伯菌、奇异紫外假单胞菌和阴沟肠杆菌中发现，这表明该基因可能在三个临床物种之间转移。结合实验使用肺炎克雷伯菌（1个分离株）、奇异紫外假单胞菌（2个分离物）和阴沟肠杆菌（2个隔离株）作为供体，大肠杆菌J53作为受体。通过PCR分析在获得的所有转导偶联物中鉴定了blaNDM-1基因。在一名患者中，四种不同的细菌携带对不同种类抗生素的耐药性基因，这大大减少了治疗选择。
Abstract
The dramatic increase in infections caused by critically multidrug-resistant bacteria is a global health concern. In this study, we characterized the antimicrobial resistance genes (ARGs) of K. pneumoniae, P. mirabilis, E. cloacae and A. baumannii isolated from both surgical wound and rectal swab of a single Italian patient. Bacterial identification was performed by MALDI-TOF MS and the antimicrobial susceptibility was carried out by Vitek 2 system. The characterization of ARGs was performed using next-generation sequencing (NGS) methodology (MiSeq Illumina apparatus). K. pneumoniae, P. mirabilis and E. cloacae were resistant to most β-lactams and β-lactam/β-lactamases inhibitor combinations. A. baumannii strain was susceptible only to colistin. The presence of plasmids (IncN, IncR, IncFIB, ColRNAI and Col (MGD2)) was detected in all Enterobacterales but not in A. baumannii strain. The IncN plasmid and blaNDM-1 gene were found in K. pneumoniae, P. mirabilis and E. cloacae, suggesting a possible transfer of this gene among the three clinical species. Conjugation experiments were performed using K. pneumoniae (1 isolate), P. mirabilis (2 isolates) and E. cloacae (2 isolates) as donors and E. coli J53 as a recipient. The blaNDM-1 gene was identified by PCR analysis in all transconjugants obtained. The presence of four different bacterial species harboring resistance genes to different classes of antibiotics in a single patient substantially reduced the therapeutic options.

https://www.mdpi.com/2079-6382/12/3/439