Federal State Autonomous Institution «National Medical Research Center of Children's Health» of the Ministry of Health.
Clinical and Research Institute of Emergency Pediatric Surgery and Trauma/CRIEPST (Publicity funded health facility of Moscow City Health Department).
Klin Lab Diagn. 2022 Mar 25;67(3):180-185. doi: 10.51620/0869-2084-2022-67-3-180-185.
Objective - assessment of RT-PCR for the detection of carbapenem-resistance genes in gram-negative bacteria. A total, 499 strains of gram-negative microorganisms isolated in two pediatric hospitals in 2019-2020 were studied. Species identification was performed using MALDI-ToF mass-spectrometry (Bruker Daltonics, Germany). Meropenem and imipenem minimal inhibitory concentration (MIC) was determined by E-test method (BioMerieux, France). The presence of acquired carbapenemase genes of IMP, NDM, VIM, KPC, OXA-48, OXA-23, OXA-40, OXA-58-groups was determined by RT-PCR. Klebsiella pneumoniae (34%), Escherichia coli (4%), Serratia marcescens (6%) and other members of Enterobacterales (6%), also gram-negative non-glucose-fermenting bacteria Acinetobacter baumannii (14%), Pseudomonas aeruginosa (36%) were found among selected strains. Carbapenemase production was found in 385 isolates (77%). The main mechanism determining carbapenem resistance in P. aeruginosa was the production of blaVIM (100%). A. baumanii strains harbored OXA-23 (55%) and OXA-40 (45%) carbapenemases. The major determinant of carbapenem resistance in K. pneumoniae isolates was OXA-48 carbapenemase, detected in 63% strains, 13% of the strains possessed blaNDM-group, 16% isolates had a combination of blaNDM-group and blaOXA-48-like. Carbapenemase of KPC-group was found in 8% K. pneumoniae strains. OXA-48 carbapenemase prevailed (95%) among S. marcescens strains. Most of E. coli isolates harbored metallo-beta-lactamase NDM (89%). Other members of Enterobacterales most often had OXA-48 carbapenemase (57%), 39% of the isolates carried blaNDM-group. In one strain, a combination of blaNDM-group and blaOXA-48-like was discovered. RT-PCR is a fast and reliable method for the detection of acquired carbapenemases and can be recommended for routine use in bacteriological laboratories.
目的 - 评估 RT-PCR 检测革兰氏阴性菌碳青霉烯耐药基因。对 2019-2020 年在两家儿科医院分离的 499 株革兰氏阴性微生物进行了研究。使用 MALDI-ToF 质谱（Bruker Daltonics，德国）进行种属鉴定。采用 E 试验法（法国生物梅里埃）测定美罗培南和亚胺培南最小抑菌浓度（MIC）。通过 RT-PCR 检测获得的 IMP、NDM、VIM、KPC、OXA-48、OXA-23、OXA-40、OXA-58 组碳青霉烯酶基因。在所选菌株中发现了肺炎克雷伯菌（34%）、大肠杆菌（4%）、粘质沙雷菌（6%）和其他肠杆菌科成员（6%），也发现了革兰氏阴性非葡萄糖发酵菌鲍曼不动杆菌（14%）、铜绿假单胞菌（36%）。在 385 株菌（77%）中发现了碳青霉烯酶的产生。铜绿假单胞菌产生碳青霉烯酶的主要机制是 blaVIM 的产生（100%）。鲍曼不动杆菌菌株携带 OXA-23（55%）和 OXA-40（45%）碳青霉烯酶。肺炎克雷伯菌分离株对碳青霉烯类药物耐药的主要决定因素是 OXA-48 碳青霉烯酶，在 63%的菌株中检测到，13%的菌株携带 blaNDM 组，16%的分离株存在 blaNDM 组和 blaOXA-48 样组合。在 8%的肺炎克雷伯菌中发现了 KPC 组的碳青霉烯酶。OXA-48 碳青霉烯酶占主导地位（95%）。大多数大肠杆菌分离株携带金属β-内酰胺酶 NDM（89%）。肠杆菌科的其他成员最常携带 OXA-48 碳青霉烯酶（57%），39%的分离株携带 blaNDM 组。在一株菌中发现了 blaNDM 组和 blaOXA-48 样的组合。RT-PCR 是一种快速可靠的检测获得性碳青霉烯酶的方法，可推荐用于细菌实验室的常规使用。
