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目的对来自住院患者血液标本分离出的1株菌株进行鉴定及碳青霉烯类抗生素耐药的基因型分析。方法利用DL-96II细菌测定系统进行菌株B1635-1的初步鉴定及药敏试验;采用PCR法扩增菌株B1635-1的16SrDNA基因序列;应用MEGA 5.0软件构建菌株B1635-1的系统发育树,确定其种属地位;采用PCR法克隆菌株B1635-1的碳青霉烯类耐药基因。结果 DL-96II细菌测定系统初步鉴定菌株B1645-1为解鸟氨酸克雷伯菌,并显示该菌株对除氨曲南以外的几乎所有β-内酰胺类抗生素耐药,对磺胺类、喹诺酮类和氨基糖苷类抗生素也耐药,但对四环素类抗生素敏感。对菌株B1645-1的16SrDNA基因序列的系统发育树分析,最终鉴定该菌为解鸟氨酸克雷伯菌(Klebsiella ornithinolytica)。菌株B1645-1扩增出编码碳青霉烯酶blaNDM-1基因,未扩增出编码碳青霉烯酶blaKPC、blaVIM、blaTME和blaSHV基因。结论首次从湖北医药学院附属东风医院住院患者的血液标本中成功分离获得一株携带blaNDM-1基因解鸟氨酸克雷伯菌株,产NDM-1型碳青霉烯酶是该株解鸟氨酸克雷伯菌耐碳青霉烯类药物的主要原因,为临床鸟氨酸克雷伯菌的感染治疗提供参考依据。鉴于肠杆菌科细菌耐药速率传播更快,警示相关部门应重视并加强对blaNDM-1基因携带菌的监测与筛查。
Objective To identify a strain isolated from blood samples of hospitalized patients and analyze the genotypes of carbapenem - resistant antibiotics. Methods The preliminary identification and drug susceptibility test of strain B1635-1 were carried out by using the DL-96II bacterial assay system. The 16S rDNA gene sequence of strain B1635-1 was amplified by PCR. The phylogenetic tree of strain B1635-1 was constructed by using MEGA 5.0 software to determine Its species status; PCR method was used to clone the carbapenem-resistant gene of strain B1635-1. Results The strain B1645-1 was initially identified as Klebsiella ornithine by DL-96II bacterial assay system and showed that this strain was resistant to almost all β-lactam antibiotics except aztreonam, Class and aminoglycoside antibiotics are also resistant, but are sensitive to tetracycline antibiotics. Phylogenetic tree analysis of the 16S rDNA gene sequence of strain B1645-1 finally identified the strain as Klebsiella ornithinolytica. Strain B1645-1 amplified the gene encoding carbapenemase blaNDM-1 without amplifying the genes encoding for carbapenemases blaKPC, blaVIM, blaTME and blaSHV. Conclusions A strain of Ornithine Ornithine isolates carrying blaNDM-1 gene was successfully isolated from blood samples of inpatients at Dongfeng Hospital, Hubei Medical College. The NDM-1 carbapenemase strain was isolated from blood samples The main reason for carbapenem-resistant carbapenem-based drugs for clinical ornithine Klebsiella infection treatment provide a reference. In view of Enterobacteriaceae bacterial resistance rate of transmission more quickly, warning the relevant departments should pay attention to and strengthen the monitoring of blaNDM-1 gene carriers and screening.