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从新生儿重症监护病房分离出的革兰氏阴性杆菌的分子特征以及超广谱β-内酰胺酶和碳青霉烯酶的表型与分子检测

Molecular Characterization of Gram-Negative Bacilli Isolated from a Neonatal Intensive Care Unit and Phenotypic and Molecular Detection of ESBL and Carbapenemase.

作者信息

Barbosa Thaís Alves, Bentlin Maria Regina, Rugolo Lígia Maria Suppo de Souza, Lyra João César, Ferreira Adriano Martison, Santos Ana Cláudia Moro Lima Dos, Teixeira Nathalia Bibiana, Medeiros Romero Letícia Calixto, Castelo Branco Fortaleza Carlos Magno, Ribeiro de Souza da Cunha Maria de Lourdes

机构信息

Department of Infectious Diseases, Botucatu Medical School, São Paulo State University (UNESP), Botucatu 18618-687, Brazil.

Department of Genetics, Microbiology and Immunology, Institute of Biosciences, São Paulo State University (UNESP), Botucatu 18618-691, Brazil.

出版信息

Antibiotics (Basel). 2025 Mar 27;14(4):342. doi: 10.3390/antibiotics14040342.

Abstract

INTRODUCTION

The increase in the rates of multidrug-resistant bacteria in healthcare environments has been recognized as a global public health problem. In view of the scarcity of data on the neonatal population, this study aimed to provide information on the genotypic and epidemiological characteristics of Gram-negative microorganisms isolated from colonization and infection sites in neonates admitted to a tertiary university center of high complexity.

METHODS

Enterobacterales and non-fermenting Gram-negative bacilli previously collected in a prospective cohort study were submitted to genotypic identification, detection of extended-spectrum β-lactamases (ESBL), carbapenemases and biofilm production, detection of specific virulence markers in , and typing by pulsed-field gel electrophoresis.

RESULTS

The data found here revealed higher rates of infection by spp. and that caused bloodstream infection and pneumonia, respectively. In this study, high biofilm production was observed, with 95.0% of Enterobacterales and 100% of non-fermenting Gram-negative bacilli being producers. Most of the isolates carried pathogenicity factors such as alginate, hemolytic phospholipase C, exotoxin A, and rhamnolipids. The phenotypic analysis of ESBL revealed that 16 (5.3%) isolates produced these enzymes. Four of these isolates (66.7%) carried the CTX-M-9 gene, three (50%) carried the TEM gene, and one (16.7%) was positive for the SHV and CMY-2 genes. Univariate and multivariate Cox regression analyses were used to identify risk factors for colonization and infection by Gram-negative microorganisms. The results of multivariate analysis revealed that biofilm production by these microorganisms was associated with the persistence of colonization by the same pathogen in the newborn and increased by 75% the daily probability of the newborn developing infection. The production of ESBL also increased the daily probability of infection by 46.8 times.

CONCLUSIONS

Enterobacterales showed average biofilm production, while the majority of non-fermenting Gram-negative bacilli were strong producers. The present data increase our knowledge of the molecular epidemiology of important Enterobacterales species, with emphasis on ESBL-producing and with emerging epidemiological potential in the neonatal intensive care unit of a tertiary university hospital. Furthermore, the results highlight the need for the monitoring and implementation of control measures and for restricting the use of broad-spectrum antibiotics.

摘要

引言

医疗环境中多重耐药菌发生率的增加已被公认为一个全球公共卫生问题。鉴于新生儿群体的数据匮乏,本研究旨在提供有关从一所高度复杂的三级大学中心收治的新生儿定植和感染部位分离出的革兰氏阴性微生物的基因型和流行病学特征的信息。

方法

将先前在前瞻性队列研究中收集的肠杆菌科细菌和非发酵革兰氏阴性杆菌进行基因型鉴定、超广谱β-内酰胺酶(ESBL)、碳青霉烯酶检测及生物膜形成检测,检测特定毒力标志物,并通过脉冲场凝胶电泳进行分型。

结果

此处发现的数据显示,分别导致血流感染和肺炎的 spp. 和 的感染率较高。在本研究中,观察到生物膜形成率较高,95.0%的肠杆菌科细菌和100%的非发酵革兰氏阴性杆菌为生物膜产生菌。大多数 分离株携带致病因子,如藻酸盐、溶血磷脂酶C、外毒素A和鼠李糖脂。ESBL的表型分析显示,16株(5.3%)分离株产生这些酶。其中4株(66.7%)携带CTX-M-9基因,3株(50%)携带TEM基因,1株(16.7%)SHV和CMY-2基因呈阳性。采用单因素和多因素Cox回归分析来确定革兰氏阴性微生物定植和感染的危险因素。多因素分析结果显示,这些微生物产生生物膜与新生儿中同一病原体定植的持续存在相关,并使新生儿发生感染的每日概率增加75%。ESBL的产生也使感染的每日概率增加46.8倍。

结论

肠杆菌科细菌显示出中等程度的生物膜形成,而大多数非发酵革兰氏阴性杆菌是强生物膜产生菌。目前的数据增加了我们对重要肠杆菌科菌种分子流行病学的认识,重点是在一所三级大学医院的新生儿重症监护病房中具有新出现流行病学潜力的产ESBL的 和 。此外,结果强调了监测和实施控制措施以及限制使用广谱抗生素的必要性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73a5/12024293/d422cbedd6e1/antibiotics-14-00342-g001.jpg

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