肺炎克雷伯菌质粒pNDM-LJ的高通量测序分析

摘要

Objective To investigate the relationship between the resistance of the Klebsiella pneumoni-ae and the Klebsiella pneumoniae plasmid pNDM-LJ carrying blaNDM-1 by high-throughput DNA sequencing. Methods High-throughput DNA sequencing was carried out by the Illumina Miseq platform , and sequencing data were assembled by Edena software. Contigs were annotated by the RAST server and analyzed by the BLAST server. Results The plasmid pNDM-LJ was 54-kb in size with a GC content of 49%. The plasmid encoded 52 putative functional genes and belonged to the IncX3 group in incompatible classifications. Analysis of the plasmid sequence revealed high similarity with other IncX3 plasmids. The blaNDM-1 gene was located in a complicated gene environment possibly constructed by several transposition events. The 5′ and 3′ ends of the blaNDM-1 gene were adjacent to the ISAba125 and IS 26 respectively , forming a 10.8-kb transposon-like structure. Conclusion The plasmid pNDM-LJ carried the blaNDM-1 gene being resistant to carbapenems and played a possibly impor-tant role in transmission of blaNDM-1 in China.%目的：通过对肺炎克雷伯菌质粒pNDM-LJ的高通量测序分析，研究其与肺炎克雷伯菌耐药的相关性。方法：利用Illumina Miseq 高通量测序平台进行测序，然后用Edena 软件拼接测序数据，再利用RAST网上工具将拼接得到的contigs 进行注释，并用BLAST网上工具进行分析。结果：pNDM-LJ 为54 kb 大小的环状质粒，GC含量约为49％，预测编码52个功能基因，在不相容群分类中属于IncX3群质粒。该质粒与已知的IncX质粒序列有较高的相似性，blaNDM-1基因所在的基因环境比较复杂，推测其是由多个转座事件共同构建的。在pNDM-LJ中，blaNDM-1基因的5′端和3′端分别与ISAba125和IS26两种插入序列相连，形成大小约为10．8 kb的转座子样结构。结论：pNDM-LJ质粒携带碳青霉烯类耐药基因blaNDM-1，可能对我国的广泛传播blaNDM-1基因发挥重要作用。