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2021 年至 2023 年期间从中国福建一家三级医院回收的耐碳青霉烯类大肠埃希菌的危险因素和分子特征。

Risk factors and molecular characterization of carbapenem resistant Escherichia coli recovered from a tertiary hospital in Fujian, China from 2021 to 2023.

机构信息

Department of Laboratory Medicine, Fujian Medical University Union Hospital, Fuzhou, Fujian, People's Republic of China.

Department of Clinical Laboratory, Jianou Hospital, Fujian, People's Republic of China.

出版信息

BMC Microbiol. 2024 Sep 28;24(1):374. doi: 10.1186/s12866-024-03525-9.

Abstract

BACKGROUND

There is a serious public health concern regarding the emergence of carbapenem-resistant Escherichia coli (CREC). The purpose of this study is to identify the molecular characterization and risk factors of CREC in Fujian province, China.

METHODS

A total of 48 CREC isolates were collected from various clinical samples. The strains were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS). Susceptibility to antibiotics was determined by the standard broth microdilution method. Polymerase chain reaction (PCR) was used to screen common drug resistance genes. Multilocus sequence typing (MLST) was used to type isolates. RT-qPCR was used to detect gene expression of acrA, acrB, and tolC. Conjugation assays were used to analyze the transferability of plasmids carrying mcr-1 or bla. Risk factors for CREC infection were identified by logistic regression analysis.

RESULTS

48 CREC strains were collected, with 81.25% producing carbapenemase (CP-CREC), and 18.75% were not producing carbapenemase (no-CP-CREC). They belonged to 21 sequence type (STs) and five unknown STs. Perianal swabs were the main sample type, with 25 patients found to have hematological malignancies. All isolates of CP-CREC were found to contain bla (bla (n = 32), bla (n = 5), bla (n = 1), and bla (n = 1)), among which one isolate co-existence bla and bla. Two bla-positive strains, specifically bla and bla, were found to co-habor mcr-1 with ST617. Conjugation assays confirmed that bla, bla, and most bla(68.75%, 22/32) could be transferred between E. coli strains. Four of the 9 non-CP-CREC isolates had deletions in ompC and ompF with bla production, while the other five showed high expression of acrA, acrB, and tolC. Antibiotics usage, antifungal treatment, detection of other pathogens (prior to CREC infection), and respiratory disease were identified as independent risk factors for CREC infection. The area under the receiver operating characteristic curve for the scoring system was 0.937. Youden's index, with sensitivity and specificity of 0.96 and 0.78, was maximal when 2 points were scored.

CONCLUSIONS

In CP-CREC, carbapenem resistance is caused primarily by multiple types of bla, while non-CP-CREC is caused by loss of porin protein or high expression of efflux pumps coupled with carrying bla. CREC isolates were highly diverse in terms of ST, with a total of 21 STs identified. Here, we first describe a clinical strain of CREC from China both mcr-1 and bla with ST617. An easy-to-use scoring system was developed to diagnose CREC infections.

摘要

背景

碳青霉烯类耐药大肠杆菌(CREC)的出现引起了严重的公共卫生关注。本研究旨在确定福建省 CREC 的分子特征和危险因素。

方法

从各种临床样本中收集了 48 株 CREC 分离株。使用基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF/MS)鉴定菌株。采用标准肉汤微量稀释法测定抗生素敏感性。聚合酶链反应(PCR)用于筛选常见的耐药基因。多位点序列分型(MLST)用于分离株分型。实时荧光定量 PCR(RT-qPCR)用于检测 acrA、acrB 和 tolC 基因的表达。接合试验用于分析携带 mcr-1 或 bla 的质粒的可转移性。通过逻辑回归分析确定 CREC 感染的危险因素。

结果

共收集了 48 株 CREC 菌株,其中 81.25%产生碳青霉烯酶(CP-CREC),18.75%不产生碳青霉烯酶(非 CP-CREC)。它们属于 21 种序列型(ST)和 5 种未知 ST。肛周拭子是主要的样本类型,25 例患者患有血液系统恶性肿瘤。所有 CP-CREC 分离株均携带 bla(bla(n=32),bla(n=5),bla(n=1)和 bla(n=1)),其中 1 株分离株同时携带 bla 和 bla。两种 bla 阳性菌株,即 bla 和 bla,与 ST617 共同携带 mcr-1。接合试验证实 bla、bla 和大多数 bla(68.75%,22/32)可在大肠杆菌菌株之间转移。9 株非 CP-CREC 分离株中有 4 株缺失 ompC 和 ompF,同时携带 bla 基因,而其余 5 株则表现出 acrA、acrB 和 tolC 的高表达。抗生素使用、抗真菌治疗、其他病原体(在 CREC 感染之前)的检测以及呼吸道疾病被确定为 CREC 感染的独立危险因素。评分系统的受试者工作特征曲线下面积为 0.937。当评分 2 分时,Youden 指数的灵敏度和特异性最高,分别为 0.96 和 0.78。

结论

在 CP-CREC 中,碳青霉烯类耐药主要由多种类型的 bla 引起,而非 CP-CREC 则由孔蛋白缺失或外排泵高表达与携带 bla 基因共同引起。CREC 分离株在 ST 方面具有高度多样性,共鉴定出 21 种 ST。在此,我们首次描述了一株携带 mcr-1 和 bla 的中国临床 CREC 菌株,与 ST617 有关。我们还开发了一种易于使用的评分系统来诊断 CREC 感染。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc8d/11438195/c568617e8beb/12866_2024_3525_Fig1_HTML.jpg

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