腹泻患者阿贡纳沙门菌耐药情况和毒力基因分析  

Antimicrobial resistance and virulence gene profiling of Salmonella Agona isolated from patients with diarrhea

作　　者：杨杰 夏琳林 赵展 刘晓霞 吴晓妹 张利娟 贾宇驰 赵妍 王玉宝 

机构地区：[1]天津医科大学第二医院感染性疾病研究所,300211

出　　处：《中华传染病杂志》2017年第11期693-698,共6页Chinese Journal of Infectious Diseases

基　　金：天津市卫生局科研基金项目（2013KZ112）

摘　　要：目的研究急性腹泻患者分离的阿贡纳沙门菌的耐药状况、分子分型和毒力基因，为了解其流行趋势和防治其感染提供依据。方法记录2013至2014年每年4月至10月天津医科大学第二医院肠道门诊急性腹泻患者的临床资料并采集患者粪便标本，进行沙门菌分离培养、生物化学鉴定、血清型分型，对鉴定得到的阿贡纳沙门菌进行抗菌药物敏感试验、脉冲场凝胶电泳分型（PFGE）、多位点序列分型（MLST）、喹诺酮耐药决定区（QRDR）、质粒介导喹诺酮耐药基因（PMQR）和β-内酰胺酶基因（TEM、SHV、OXA和CTX-M）的PCR扩增与测序、沙门菌毒力岛（SPI）1～6、9～12和毒力质粒代表性基因的PCR检测，并分析阿贡纳沙门菌感染患者的临床特点。结果2年共检测到非重复非伤寒沙门菌119株，其中8株（6.7%）为阿贡纳沙门菌。阿贡纳沙门菌的耐药率依次为链霉素100.0%，氨苄西林和庆大霉素62.5%，萘啶酸、环丙沙星和左氧氟沙星25.0%，氯霉素、阿莫西林/克拉维酸钾和哌拉西林/他唑巴坦12.5%，对其余药物均敏感。阿贡纳沙门菌MLST分型均为ST13，PFGE显示5个克隆，有4株为同一克隆并且药敏表型完全相同。2株氟喹诺酮类（FQ）抗菌药物耐药株都出现GyrA亚基87位氨基酸密码子突变，未检出PMQR耐药基因，但其中1株环丙沙星K-B法显示敏感。5株氨苄西林耐药株都携带TEM-1b基因，其中1株对β-内酰胺/β-内酰胺酶抑制剂复合物耐药。全部阿贡纳沙门菌都扩增出SPI1～6、9、11、12的代表性基因hilA、sseL、mgtC、siiE、sopB、pagN、bapA、pagC和sspH2，未检出SPI10代表性基因sefA和毒力质粒基因spvB、prot6E。2例FQ耐药株感染者临床诊断为细菌性痢疾，其余6例FQ敏感株感染者临床诊断为急性胃肠炎。结论临床分离的阿贡纳沙门菌出现氟喹诺酮耐药和多重耐药，携带多种毒力基因，菌株对FQ的耐药性和患�ObjectiveTo investigate the antimicrobial resistance, molecular phenotypes, virulence gene profiles of Salmonella Agona （S.Agona） isolated from patients with acute diarrhea, and to better understand its epidemic trend, prevention and treatment.MethodsClinical data and stool samples of patients with acute diarrhea during April to October in 2013 and 2014 from the Second Hospital of Tianjin Medical University were collected. Enrichment culture, biochemical identification and serotyping analysis were used to isolate and identify S. Agona strains. The isolated strains were further analyzed with antibiotics susceptibility test, pulsed field gel electrophoresis （PFGE）, multiple locus sequence typing （MLST）, Quinolone resistance determining region （QRDR）. Plasmid-mediated quinolone resistance （PMQR） and β-lactamases genes （TEM, SHV, OXA, and CTX-M） were amplified by polymerase chain reaction （PCR） and sequencing. The representative genes carried by Salmonella pathogenicity islands （SPI） 1—6, 9—12 and virulence plasmids were amplified by PCR. And the clinical characteristics of S. Agona infection were analyzed.ResultsAmong 119 non-repetitive （non-typhoidal salmonella, NTS） isolates during the two years, eight isolates （6.7%） of S. Agona were identified. The resistance rate of S. Agona strains to streptomycin was 100.0%, those to ampicillin and gentamicin were 62.5%, to levofloxacin, ciprofloxacin and nalicixic acid were 25.0%, to chloramphenicol, amoxillin/clavulanic acid and piperacillin tazobactam were 12.5%. The strains were susceptible to other drugs. All 8 isolates had the identical ST13 genotype. PFGE showed 5 clones, and 4 out of 5 isolates had the exact same patterns of PFGE and drug susceptibility. Two （fluoroquinolones, FQ） resistant strains carried gyrA mutation leading to amino acid substitutions at position 87 in GyrA, and no PMQR genes was detected, while one of which was sensitive to ciprofloxacin by K-B method. All five ampicillin-resistant isolates were po

关 键 词：阿贡纳沙门菌 抗菌药物耐药性 多位点序列分型 脉冲场凝胶电泳 感染暴发 沙门菌毒力岛 

分 类 号：R446.5[医药卫生—诊断学] R574.62[医药卫生—临床医学]