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2017年9月至2018年5月云南省某院新生儿肠道耐药肠杆菌目细菌筛查及耐药性分析

Screening and drug resistance analysis of enterobacterales in neonatal gut microbiota in a hospital in Yunnan province, from December 2017 to May 2018

    摘要
  • 摘要:
    目的  了解产科病房新生儿肠道中耐药肠杆菌目细菌携带状况及菌株特征。
    方法  收集2017年9月至2018年5月间云南省某医院产科病房新生儿粪便样本277份，通过超广谱 β-内酰胺酶（ESBL） 显色培养基分离培养、菌株药物敏感检测及代表菌株Illumina基因组测序，分析新生儿肠道耐药菌携带状况，菌株耐药表型、耐药基因型及序列分型（STs）特征。
    结果  277份样品中113份样品ESBL显色培养基培养阳性，阳性率40.79%。 分离鉴定出4个属11个种的113株肠杆菌目细菌，其中，克雷伯属53.10%（60/113），埃希菌属30.10%（34/113），肠杆菌属15.04%（17/113）及勒克氏菌属1.77%（2/113）。 菌株呈现多重耐药性，除对二代、三代头孢菌素高度耐药外，对其他种类代表抗菌药物耐药率分别为厄他培南52.21%（59/113）、甲氧苄啶/磺胺甲恶唑72.57%（82/113）、四环素65.49%（74/113）和环丙沙星54.87%（62/113）。 挑选84株代表菌株完成基因组序列分析，共筛查到70种耐药基因，携带率较高的有sul1（84.52%，71/84）、fosA（71.43%，60/84）、tet(A)（67.86%，57/84）、blaSHV（63.10%，53/84）和blaNDM（61.90%，52/84）基因。 53株肺炎克雷伯菌呈现优势克隆型集中分布，ST17型占比达92.45%（49/53），且均携带blaNDM-1基因，其他种属菌株ST型别呈多样性。 检出携带blaNDM-1的阴沟肠杆菌、霍氏肠杆菌，携带blaIMP-26的格氏克雷伯菌，携带blaIMP-1的勒克氏菌和携带mcr的阿氏肠杆菌、霍氏肠杆菌及赫氏埃希菌。
    结论  本研究发现了某产科病房新生儿肠道高比例携带ESBL耐药肠杆菌，揭示了耐药菌株极易在新生儿间传播和肠道定植，应高度重视新生儿耐药菌环境暴露风险及对其肠道微生态建立的影响。

     

    Abstract:
    Objective  To investigate the colonization status and strain characteristics of resistant Enterobacterales in the intestine of neonates.
    Methods  277 fecal samples of neonates in the maternity ward of a hospital were collected from September 2017 to May 2018 in Yunnan province. We analyzed the status of carriage of drug-resistant bacteria in the intestinal tract of neonates, strain resistance phenotypes, resistance genotypes and sequence types (STs) characteristics by isolation and culture in extended spectrum β-lactamase (ESBL) chromogenic medium, strain drug susceptibility testing and genome sequencing of Illumina, a representative strain.
    Results  113 fecal samples out of 77 samples were ESBL-positive strains, with a positivity rate of 40.79%. 113 Enterobacterales strains of 11 species from 4 genera were isolated and identified, including Klebsiella 53.10% (60/113), Escherichia 30.10% (34/113), Enterobacter 15.04% (17/113), and Leuconostoc 1.77% (2/113). The strains presenting multidrug-resistant profiles. Except for highly resistant to second and third-generation cephalosporin, the resistance rates of carbapenems, sulfonamides, tetracyclines and ciprofloxacin were 52.21% (59/113), 72.57% (82/113), 65.49% (74/113), and 54.87% (62/113), respectively. The genome sequences of 84 representaive strains were screened, and al of 70 drug r(esis)tance genes were detected, including sul1 (84.52%, 71/84), fosA (71.43%, 60/84), tet(A) (67.86%, 57/84), blaSHV (63.10%, 53/84) and blaNDM (61.90%, 52/84). 53 Klebsiella pneumoniae showed a concentrated distribution of clones, ST17 carrying blaNDM-1 gene accounted for 92.45% (49/53). The STs of other species were diverse. Enterobacter cloacae and Enterobacter hormaechei carrying blaNDM-1, Klebsiella gordonii carrying blaIMP-26, Leclercia carrying blaIMP-1, and Enterobacter albus, E. hormaechei and Escherichia hermannii carrying mcr were detected.
    Conclusion  It is found that a high proportion of neonates in a maternity ward carried ESBL-resistant enterobacters in their intestines, revealing that drug-resistant strains are highly susceptible to inter-neonatal transmission and intestinal colonization. Attention should be paid to the environmental exposure risk of neonatal drug-resistant bacteria and its impact on the establishment of intestinal microecology.

     

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