Ou Weimei, Cui Lanqing, Li Yun, Zheng Bo, Lv Yuan
The Institute of Clinical Pharmacology, Peking University First Hospital, Peking University, Beijing, 100191, P.R. China.
PLoS One. 2014 Dec 3;9(12):e113852. doi: 10.1371/journal.pone.0113852. eCollection 2014.
The study aimed to investigate the prevalence and epidemiological characteristics of blaNDM-1 (encoding New Delhi metallo-β-lactamase 1) in Enterobacteriaceae and the Acinetobacter calcoaceticus-Acinetobacter baumannii complex (ABC) in China from July 2011 to June 2012.
PCR was used to screen for the presence of blaNDM-1 in all organisms studied. For blaNDM-1-positive strains, 16S rRNA analysis and Analytical Profile Index (API) strips were used to identify the bacterial genus and species. The ABCs were reconfirmed by PCR detection of blaOXA-51-like. Antibiotic susceptibilities of the bacteria were assessed by determining minimum inhibitory concentration (MIC) of them using two-fold agar dilution test, as recommended by the Clinical and Laboratory Standards Institute (CLSI). Molecular typing was performed using pulsed-field gel electrophoresis (PFGE). S1 nuclease-pulsed-field gel electrophoresis (S1-PFGE) and Southern blot hybridization were conducted to ascertain the gene location of blaNDM-1. Conjugation experiments were conducted to determine the transmission of blaNDM-1-positive strains.
Among 2,170 Enterobacteriaceae and 600 ABCs, seven Enterobacteriaceae strains and two A. calcoaceticus isolates from five different cities carried the blaNDM-1 gene. The seven Enterobacteriaceae strains comprised four Klebsiella pneumoniae, one Enterobacter cloacae, one Enterobacter aerogen and one Citrobacter freundii. All seven were non-susceptible to imipenem, meropenem or ertapenem. Two A. calcoaceticus species were resistant to imipenem and meropenem. Three K. pneumoniae showed the same PFGE profiles. The blaNDM-1 genes of eight strains were localized on plasmids, while one was chromosomal.
Compared with previous reports, the numbers and species containing the blaNDM-1 in Enterobacteriaceae have significantly increased in China. Most of them are able to disseminate the gene, which is cause for concern. Consecutive surveillance should be implemented and should also focus on the dissemination of blaNDM-1 among gram-negative clinical isolates.
本研究旨在调查2011年7月至2012年6月间中国肠杆菌科细菌及醋酸钙不动杆菌-鲍曼不动杆菌复合体(ABC)中blaNDM-1(编码新德里金属β-内酰胺酶1)的流行情况及流行病学特征。
采用聚合酶链反应(PCR)筛选所有研究菌株中blaNDM-1的存在情况。对于blaNDM-1阳性菌株,使用16S核糖体RNA(rRNA)分析和分析谱指数(API)条带鉴定细菌属和种。通过PCR检测blaOXA-51样基因对ABC进行重新确认。按照临床和实验室标准协会(CLSI)的建议,采用二倍稀释琼脂试验测定细菌的最低抑菌浓度(MIC),评估细菌的抗生素敏感性。使用脉冲场凝胶电泳(PFGE)进行分子分型。进行S1核酸酶脉冲场凝胶电泳(S1-PFGE)和Southern印迹杂交以确定blaNDM-1的基因位置。进行接合试验以确定blaNDM-1阳性菌株的传播情况。
在2170株肠杆菌科细菌和600株ABC中,来自五个不同城市的7株肠杆菌科菌株和2株醋酸钙不动杆菌分离株携带blaNDM-1基因。7株肠杆菌科菌株包括4株肺炎克雷伯菌、1株阴沟肠杆菌、1株产气肠杆菌和1株弗氏柠檬酸杆菌。所有7株对亚胺培南、美罗培南或厄他培南均不敏感。2株醋酸钙不动杆菌对亚胺培南和美罗培南耐药。3株肺炎克雷伯菌显示相同的PFGE图谱。8株菌株的blaNDM-1基因定位于质粒上,1株定位于染色体上。
与以往报道相比,中国肠杆菌科中携带blaNDM-1的数量和种类显著增加。其中大多数能够传播该基因,令人担忧。应实施连续监测,并应关注blaNDM-1在革兰氏阴性临床分离株中的传播。