高通量全基因组测序法对尿道致病性大肠埃希菌NB8株毒力因子数据的初步分析

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摘要目的：调查1株尿道致病性大肠埃希菌（UPEC）NB8株携带的毒力因子和可能存在的致病机制。方法：UPEC NB8株分离自2012年4月宁波市第一医院住院患者尿液标本，做16SrDNA和gyrA基因测序BLASTn比对确认为UPEC，用Illumina HiSeq与Ion Torrent PGM这2种大规模并行测序仪进行全基因组分析，再进行人工测序补缺口。再对NB8株做毒力因子数据挖掘和分析，最后把NB8株和9株全基因组测序的UPEC的毒力因子做比较基因组学研究。结果：UPEC NB8株全基因组测序得到一条推定的染色体序列，长4 550 369 bp（内含14个缺口）。推定的质粒序列，长635 377 bp（内含33个缺口）。NB8株全基因组中挖掘出黏附（黏附素、P菌毛、1型菌毛、大肠埃希菌共同菌毛）、铁摄取系统（产气菌素、血红素摄取、肠杆菌素）、蛋白酶（分泌自转运毒素）、毒素（溶血素）、抗原43、SHI-2毒力岛等多种毒力因子。结论：UPEC NB8株中检出的多种毒力因子可能会在细菌黏附、侵袭、溶细胞活性、细胞毒性等方面起着重要作用，并能增强细菌的生存能力和致病力，导致宿主的泌尿道感染。全基因组测序技术具有超高速、高通量、低成本和高效益的优势，值得推广。

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关键词 ：大肠埃希菌, 全基因组, 多药耐药, 尿道致病性, 毒力因子

Abstract：Objective: To investigate virulence factors and pathogenic mechanisms in uropathogemic Escherichia coli (UPEC) NB8. Methods: UPEC NB8 was collected from urine sample of an inpatient in Ningbo First Hospital, in April, 2012. Sequencing gyrA and parC, BLASTn algorithm were performed to identify E.coli. Then, whole genome were sequenced by Illumina HiSeq and Ion Torrent PGM, and PCR was performed to fill gaps. Data mining of virulence factors were performed, and comparative genomics of virulence factors in NB8 and 9 strains of UPEC after whole genome sequencing were performed. Results: By whole genome sequencing, a putative chromosome sequence (14 gaps inside) was 4 550 369 bp, a putative plasmid sequence (33 gaps inside) was 635 377 bp. Some classes of virulence factors were positive in NB8, including adhesion (adhesin, type P fimbriae, type I fimbriae, Escherichia coli common pilus), iron uptake(aerobactin, heme, enterobactin), protease (secreted autotransporter toxin), toxin (hemolysin), Ag43, SHI-2 pathogenicity island protein. Conclusion: Virulence factors in UPEC NB8 played an important role in adhesion, invasion, cell viability, cell toxicity, and contribute to enhance the survival and virulence of the bacteria, so lead to urinary tract infection. Whole genome sequencing technology has the advantages of high speed, high throughput, low cost and high efficiency, which is worth of promoting.

Key words： Escherichia coli whole genome multi-drug resistance uropathogemic virulence factor

收稿日期: 2016-01-27

基金资助:宁波市自然科学基金资助项目（2012A610186）；浙江省中医药资助项目（2011ZB126）。

作者简介: 翁幸鐾（1980-），男，浙江宁波人，副主任技师。

链接本文:

https://xb.wmu.edu.cn/CN/ 或 https://xb.wmu.edu.cn/CN/Y2016/V46/I11/798

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