目的通过高通量测序法对多重耐药大肠埃希菌HX43进行耐药分子机制的研究。方法用Illumina Miseq平台对HX43进行高通量测序,用Edena、RAST、ResFinder、MLST和BLAST等生物信息学工具或数据库进行数据分析,获得耐药基因相关序列数据。结果HX43对多种临床常用抗生素均不敏感,仅对碳氢霉烯类药物敏感。对高通量测序数据的分析研究发现,该菌存在多种耐药基因,包括β-内酰胺类耐药基因3个(blacmy-42、blaCTX-M-14和blaOXA-30),氨基糖苷类耐药基因5个(aac(3)-Ila、aadA5、strA、strB和aaC(6’)-Ib—cr),喹诺酮类耐药基因1个(aoA7(6’)-Ib—cr),磺胺及甲氧苄啶类耐药基因3个(sul1、sul2和dfrA17),四环素耐药基因1个(tet(B)),氯霉素耐药基因2个(catB3和cmlA1),大环内酯类耐药基因2个(erm(B)和mph(A))。对包含blaCMY-42的contigs进行分析,发现该基因与ISEcp1插入序列、blc和sugE等基因相关联。质粒分型发现HX43携带5种不相容群的质粒。多位点序列分型(MLST)分析发现HX43属于ST3835,为国内外较少见的序列型。结论高通量测序技术可准确获得临床菌株抗生素耐药的相关基因信息,为临床抗菌治疗提供重要的实验室数据支持。
Objective To investigate the molecular resistance mechanism of Escherichia coli HX43 by high- throughput sequencing. Methods HX43 was sequenced by the Illumina Niseq platform, and sequencing data were analyzed by the Edena, RAST, ResFinder, MLST and BLAST softwares and databases. Results HX43 was resistant to most common clinical antibiotics except carbapenems. Analysis of data revealed resistance genes to β-1actams ( blaCMY-42 , blaCTX-M-14 and blaOXA-30 ) , aminoglycosides ( aac (3) -Ila, aadA5 , strA , strB and aac (6') -Ib-cr ) , quinolones ( aac (6') -Ib-cr ) , trimethoprim/sulfonamides (sul1, sul2 and dfrA17 ) , tetracyclines (tet ( B ) ) , chloramphenicol (catB3 and cmlA1 ) , macrolides (erm (B) and mph (A)). Sequence analysis of the contig containing blaCMY-42 identified correlations of the gene with ISEcpl insertion sequences, blc and sugE genes. Plasmid typing identified 5 plasmid incompatibility groups in HX43. MLST analysis found that HX43 belonged to ST3835, a relatively rare sequence type in the world. Conclusion Information of resistance genes can be obtained by high-throughput sequencing, which provides important experimental data for clinical antimicrobial treatment.