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  

Data analysis of virulence factors in uropathogemic Escherichia coli NB8 by whole genome sequencing

作　　者：翁幸鐾[1] 糜祖煌[2] 王春新[3] 朱健铭 WENG Xingbei;MI Zuhuang;WANG Chunxin;ZHU Jianming(Department of Medical Laboratory,Ningbo First Hospital, Ningbo, 315010;Department of Bioinformation, Wuxi Clon-Gen Techonology Institute,Wuxi, 214026;Department of Medical Laboratory, Wuxi People’s Hospital Affiliated to Nanjing Medical University,Wuxi, 214023;;Department of Medical Laboratory, Hangzhou Yuhang Hospital of Traditional Chinese Medicine, Hangzhou, 311106)

机构地区：[1]宁波市第一医院检验科,浙江宁波315010 [2]无锡市克隆遗传技术研究所生物信息学室,江苏无锡214026 [3]南京医科大学附属无锡人民医院检验科,江苏无锡214023 [4]杭州市余杭区中医院检验科,浙江杭州311106

出　　处：《温州医科大学学报》2016年第11期798-806,共9页Journal of Wenzhou Medical University

基　　金：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)

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16Sr DNA和gyr A基因测序BLASTn比对确认为UPEC,用Illumina Hi Seq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。Objective: To investigate virulence factors and pathogenic mechanisms in uropathogemicEscherichia coli (UPEC) NB8. Methods: UPEC NB8 was collected from urine sample of an inpatient in NingboFirst 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 fillgaps. Data mining of virulence factors were performed, and comparative genomics of virulence factors in NB8and 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, typeP 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, andcontribute to enhance the survival and virulence of the bacteria, so lead to urinary tract infection. Whole genomesequencing technology has the advantages of high speed, high throughput, low cost and high efficiency, which isworth of promoting.

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 

分 类 号：R378.2[医药卫生—病原生物学]