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| 临床分离弗劳地枸橼酸杆菌耐药性分析及质粒介导的AmpC酶基因检测 |
| Antimicrobial resistance analysis and detection of plasmid-mediated AmpC β-lactamase genes in clinical isolates of Citrobacter freundii |
| 投稿时间：2019-02-25 |
| DOI：10.3969/j.issn.1000-0399.2019.09.002 |
| 中文关键词: 弗劳地枸橼酸杆菌 细菌耐药性 β-内酰胺酶 AmpC酶 质粒 |
| 英文关键词: Citrobacter freundii Bacteria resistance Beta-lactamase AmpC Plasmid |
| 基金项目:国家自然科学基金项目（项目编号：81673242） |
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| 中文摘要: |
| 目的 了解安徽地区临床分离弗劳地枸橼酸杆菌临床分布、耐药性及质粒介导AmpC酶（pAmpCs）基因的流行情况。方法 收集2012~2017年安徽省细菌耐药监控中心监测网内的34所医院报送的临床分离的104株弗劳地枸橼酸杆菌，采用琼脂稀释法进行抗生素敏感性实验。提取细菌质粒，聚合酶链式反应（PCR）检测质粒介导AmpC酶基因，对阳性扩增产物进行测序分析；AmpC酶阳性菌株做转移接合试验，PCR扩增并测序确定接合子的基因型；测定供体菌、受体菌和接合子对多种抗生素的最低抑菌浓度（MIC）。结果 104株弗劳地枸橼酸杆菌临床分布广泛，最多见于呼吸内科（30.77%）及重症监护病房（24.04%），标本来源以痰液、尿液最多。4株AmpC酶基因阳性的菌株中，有3株DHA-1基因阳性，1株ACT-1基因阳性。4株AmpC酶基因阳性的菌株有3株转移接合成功。这3株接合子与受体菌相比，对多种常用抗生素尤其是β-内酰胺类抗生素的MIC值提高了8~128倍。结论 弗劳地枸橼酸杆菌临床分布广泛，耐药严重，携带有质粒介导的AmpC酶基因的菌株对多种β-内酰胺类抗生素耐药，并且可通过接合性质粒在不同菌属间播散。 |
| 英文摘要: |
| Objective To investigate the clinical distribution, antimicrobial resistance and prevalence of pAmpCs genes in clinical isolates of Citrobacter freundii in Anhui. Methods The minimum inhibition concentration (MIC) of 104 strains of Citrobacter freundii collected from various clinical specimens from 34 hospitals in the monitoring network of bacterial resistance monitoring center in Anhui province during 2012 to 2017 were determined by agar dilution method. The AmpC β-lactamase genes were screened by polymerase chain reaction (PCR) in all the strains and the positive isolates were subsequently confirmed by sequencing. The conjugation experiments were performed for AmpC β-lactamase genes positive strains. The MICs of recipient strains and successful conjugants in the conjugation experiments were also tested by agar dilution method for various antimicrobial agents. Results A total of 104 stains of Citrobacter freundii were widely distributed, most of which presented in the Department of Respiratory Medicine and ICU, and sputum and urine were the most common sources of specimens. Among the 4 strains of positive AmpC genes, 3 strains harboured DHA-1 gene and 1 strain harboured ACT-1 gene. Three of the 4 AmpC gene positive strains were successfully transferred and ligated. The MICs of conjugants for many antibiotics especially beta-lactam antibacterial drugs were up-regulated by 8 times to 128 times, compared to recipient strains. Conclusions Citrobacter freundii is widespread and can be isolated from various specimens. Strains carrying pAmpC genes are resistant to a variety of β-lactamase antibiotics and can be disseminated among different bacteria by conjugation plasmids. |
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