三级医院环境中机会性病原体和抗生素抗性基因的图谱绘制

Cartography of opportunistic pathogens and antibiotic resistance genes in a tertiary hospital environment

作者信息Kern Rei Chng, Chenhao Li, Denis Bertrand, Amanda Hui Qi Ng, Junmei Samantha Kwah, Hwee Meng Low, Chengxuan Tong, Maanasa Natrajan, Michael Hongjie Zhang, Licheng Xu, Karrie Kwan Ki Ko, Eliza Xin Pei Ho, Tamar V Av-Shalom, Jeanette Woon Pei Teo, Chiea Chuen Khor, MetaSUB Consortium, Swaine L Chen, Christopher E Mason, Oon Tek Ng, Kalisvar Marimuthu, Brenda Ang, Niranjan Nagarajan, David Danko, Daniela Bezdan, Ebrahim Afshinnekoo, Sofia Ahsanuddin, Chandrima Bhattacharya, Daniel J Butler, Kern Rei Chng, Francesca De Filippis, Jochen Hecht, Andre Kahles, Mikhail Karasikov, Nikos C Kyrpides, Marcus H Y Leung, Dmitry Meleshko, Harun Mustafa, Beth Mutai, Russell Y Neches, Amanda Ng, Marina Nieto-Caballero, Olga Nikolayeva, Tatyana Nikolayeva, Eileen Png, Jorge L Sanchez, Heba Shaaban, Maria A Sierra, Xinzhao Tong, Ben Young, Josue Alicea, Malay Bhattacharyya, Ran Blekhman, Eduardo Castro-Nallar, Ana M Cañas, Aspassia D Chatziefthimiou, Robert W Crawford, Youping Deng, Christelle Desnues, Emmanuel Dias-Neto, Daisy Donnellan, Marius Dybwad, Eran Elhaik
PMID32514171
期刊Nat Med
发布时间2020-06
DOI10.1038/s41591-020-0894-4

摘要

尽管消毒是感染控制的关键环节,医院环境微生物的定植模式和耐药基因组仍未得到充分探索。我们通过对45张病床相关179个位点进行重复采样(间隔最长1.5年),首次对某三级医院的微生物组、病原体和抗生素耐药基因盒进行了大规模基因组表征。深度鸟枪法宏基因组学揭示了微生物与抗生素耐药基因的独特生态位特征,这些位点以生物膜形成环境和人类微生物组影响环境为特征,并呈现相应的时空分异模式。基于纳米孔测序的准宏基因组学技术获得了数千个高连续性基因组、噬菌体和质粒序列(>60%为新发现),实现了对医院环境中耐药基因组和可移动遗传元件多样性及动态结构的系统表征。系统发育分析表明多重耐药菌株在医院环境中广泛分布且能稳定定植。与临床分离株的比对显示此类微生物可在医院环境中长期存留(>8年),伺机感染患者。这些发现凸显了医院抗生素耐药库表征的重要性,并证实了通过系统性监测优化感染防控资源配置的可行性。

实验方法

产品清单

名称品牌货号
Isohelix DNA口腔拭子IsohelixSK-4S
DNA/RNA保护液Zymo ResearchZYR.R1100-250
裂解基质E管MP Biomedicals116914500
FastPrep-24仪器----
Maxwell RSC血液DNA提取试剂盒PromegaAS1400
Qubit 2.0荧光计----
Qubit双链DNA高灵敏度检测试剂盒Life TechnologiesQ32854
CovarisCovaris--
NEBNext超DNA试剂盒New England BiolabsE7370
Illumina HiSeq2500平台IlluminaHiSeq2500
NEBNext超II DNA文库制备试剂盒New England BiolabsE7645
Illumina HiSeq 4000平台IlluminaHiSeq4000
脑心浸液肉汤Thermo Scientific MicrobiologyCM1135B
Maxwell RSC培养细胞试剂盒PromegaAS1620
Agencourt AMPure XP磁珠Beckman CoulterA63882
NanoDrop----
1D²测序试剂盒--SQK-LSK308
1D测序试剂盒--SQK-LSK108
1D测序试剂盒--SQK-LSK109
天然条形码试剂盒--EXP-NBD103
天然条形码试剂盒--EXP-NBD104
天然条形码试剂盒--EXP-NBD114
MIN106流通池--MIN106
MIN106D流通池--MIN106D
MIN107流通池--MIN107
GridION测序仪----
0.5%琼脂糖凝胶----