首页 >>2014年06期
山东省O139群霍乱弧菌的分子特征及耐药性研究
作者:袁玉起 吕慧 周海健 崔志刚 孙娜 关冰 邵坤 毕振旺 阚飙 毕振强

摘要:

目的  调查山东省O139群霍乱弧菌分子流行病学特征及耐药谱。方法  对山东省自1997年O139群霍乱疫情出现以来分离的13株O139群霍乱弧菌(9株为临床分离株,4株为环境株)进行生化鉴定及血清型复核,同时进行PFGE分子分型和毒力基因及耐药相关基因检测,使用微量肉汤稀释法检测其耐药性。结果  山东省13株O139群霍乱弧菌聚类为7个PFGE型别,其中2013年济宁的1株临床株和2株环境株聚为KZGN11O139.CN0077,2005年济南的1株临床株、2005年济宁的1株临床株和2009年菏泽的1临床株聚为KZGN11O139.CN0002,其余均各自为不同的型别。2株烟台临床株缺失ctxABtcpI基因,PFGE条带也较为特殊,其他11株霍乱弧菌均含有hlyActxABtoxRtcpIrtxAtcpA 6种毒力基因。13株霍乱弧菌均含有耐药相关基因intI 1、intI 4及sxt。在12种抗生素中,卡那霉素、复方磺胺甲恶唑、氨苄西林及庆大霉素的耐药率分别为11/13、9/13、7/13和7/13。结论  山东省O139群霍乱弧菌PEGE分型提示菌株间可能存在流行病学关联,多数菌株产毒且耐药。

关键词:弧菌,霍乱;电泳,凝胶,脉冲场;毒力;耐药

Abstract:

Objective  To investigate the molecular epidemiological characteristics and antibiotic resistance profiles of Vibrio cholerae O139 in Shandong province.Methods  A total of 13 strains of V. cholerae O139 (9 clinical strains and 4 environmental strains) isolated from cholera epidemics in Shandong province since 1997 were recovered and confirmed with serum agglutination and biochemical reaction. Pulsed-field gel electrophoresis (PFGE) was carried out for molecular subtyping. Virulence genes and drug resistance related genes were detected by PCR. Antibiotic susceptibility tests were performed using micro-broth dilution method.Results  Thirteen strains of V. cholerae O139 were differentiated into seven pulsetypes. One clinical strain and two environmental strains isolated from Jining in 2013 were clustered into the pulsetype namely KZGN11O139. CN0077, and an identical PFGE pattern of KZGN11O139. CN0002 was found among three clinical strains from Jinan in 2005, Jining in 2005 and Heze in 2009. Other pulsotypes were unique in China and found only in Shandong province. Because of deletion of ctxAB and tcpI, the PFGE patterns of two strains isolated from Yantai in 2000 and 2004 were different from other 11 strains which harbored ctxAB, tcpA, tcpI, rtxA, hlyA and toxR. All strains contained one or more drug resistance related genes such as intI 1, intI 4 and sxt, and were resistant to two kinds of antibiotics at least. Among the 12 kinds of antibiotics, the resistant ratioes to kamamycin, trimethoprim-sulfamethoxazole, ampicillin and gentamicin were 11/13, 9/13, 7/13 and 7/13, respectively.Conclusion  Molecular subtyping indicates possible epidemiological links among V.cholerae O139 in Shandong province, and almost all strains were toxigenic and drug resistant.

Key words: Vibrio cholerae;Electrophoresis,gel,pulsed-field;Virulence;Antibiotic resistance

发表日期:2014/6

引用本文:

图/表:

  • 10.3760/cma.j.issn.0253-9624.2014.06.007.F001

    10.3760/cma.j.issn.0253-9624.2014.06.007.F001

  • 10.3760/cma.j.issn.0253-9624.2014.06.007.T001

    10.3760/cma.j.issn.0253-9624.2014.06.007.T001

  • 10.3760/cma.j.issn.0253-9624.2014.06.007.T002

    10.3760/cma.j.issn.0253-9624.2014.06.007.T002

  • 10.3760/cma.j.issn.0253-9624.2014.06.007.T003

    10.3760/cma.j.issn.0253-9624.2014.06.007.T003

参考文献:

[1]肖东楼. 霍乱防治手册[M].6版. 北京:人民卫生出版社, 2013.
[2]CooperKL, LueyCK, BirdM, et al. Development and validation of a PulseNet standardized pulsed–field gel electrophoresis protocol for subtyping of Vibrio cholerae[J]. Foodborne Pathog Dis, 2006, 3(1):51–58.
[3]TehCS, ChuaKH, ThongKL. Genetic variation analysis of Vibrio cholerae using multilocus sequencing typing and multi–virulence locus sequencing typing[J]. Infect Genet Evol, 2011, 11(5):1121–1128.
[4]LiM, ShimadaT, MorrisJGJr, et al. Evidence for the emergence of non–O1 and non–O139 Vibrio cholerae strains with pathogenic potential by exchange of O–antigen biosynthesis regions[J]. Infect Immun, 2002, 70(5):2441–2453.
[5]Shuan Ju TehC, Lin ThongK, Tein NgoiS, et al. Molecular characterization of serogrouping and virulence genes of Malaysian Vibrio cholerae isolated from different sources[J]. J Gen Appl Microbiol, 2009, 55(6):419–425.
[6]ChowKH, NgTK, YuenKY, et al. Detection of RTX toxin gene in Vibrio cholerae by PCR[J]. J Clin Microbiol, 2001, 39(7):2594–2597.
[7]RiveraIN, ChunJ, HuqA, et al. Genotypes associated with virulence in environmental isolates of Vibrio cholerae[J]. Appl Environ Microbiol, 2001, 67(6):2421–2429.
[8]WangR, LouJ, LiuJ, et al. Antibiotic resistance of Vibrio cholerae O1 El Tor strains from the seventh pandemic in China, 1961–2010[J]. Int J Antimicrob Agents, 2012, 40(4):361–364.
[9]ShiL, FujiharaK, SatoT, et al. Distribution and characterization of integrons in various serogroups of Vibrio cholerae strains isolated from diarrhoeal patients between 1992 and 2000 in Kolkata, India[J]. J Med Microbiol, 2006, 55(Pt 5):575–583.
[10]BhanumathiR, SabeenaF, IsacSR, et al. Molecular characterization of Vibrio cholerae O139 bengal isolated from water and the aquatic plant Eichhornia crassipes in the River Ganga, Varanasi, India[J]. Appl Environ Microbiol, 2003, 69(4):2389–2394.
[11]Clinical and Laboratory Standards Institute.CLSI Documents M100–S20 Performance standards for antimicrobial susceptibility testing;Twentieth informational supplement[S]. Wayne PA:CLSI, 2010.
[12]古文鹏, 伏晓庆, 杨建斌, 等. 2011年云南省中缅边境地区三起霍乱疫情菌株分子分型研究[J]. 中华预防医学杂志, 2012, 46(6):573–574.
[13]张宝, 周健, 徐相凯. 山东省潍坊市霍乱疫情五十年回顾[J]. 医学动物防制, 2004, 20(7):444–446.
[14]姜玉芳, 刘宗东, 陈晓君, 等. 山东省莱州市首例0139霍乱病例监测报告[J]. 中华流行病学杂志, 2005, 26(6):407.
[15]程晋霞, 曾静, 刘莉, 等. 海产品中霍乱弧菌免疫磁分离和实时荧光PCR快速检测方法的建立[J]. 中华预防医学杂志, 2014, 48(2):133–137.
[16]GoeringRV. Pulsed field gel electrophoresis:a review of application and interpretation in the molecular epidemiology of infectious disease[J]. Infect Genet Evol, 2010, 10(7):866–875.
[17]严寒秋, 李伟, 吴疆, 等. 一起由非产毒霍乱弧菌引发腹泻暴发的调查[J]. 中华流行病学杂志, 2006, 27(10):918–919.
[18]Rowe–MagnusDA, GueroutAM, MazelD. Bacterial resistance evolution by recruitment of super–integron gene cassettes[J]. Mol Microbiol, 2002, 43(6):1657–1669.
[19]李彦媚, 赵喜红, 徐泽智, 等. 新型细菌耐药元件––整合子系统[J]. 中国抗生素杂志, 2012, 37(1):1–7.
[20]HochhutB, MarreroJ, WaldorMK. Mobilization of plasmids and chromosomal DNA mediated by the SXT element, a constin found in Vibrio cholerae O139[J]. J Bacteriol, 2000, 182(7):2043–2047.
[21]于礼, 王多春, 阚飙, 等. 霍乱弧菌中SXT整合性接合元件研究进展[J]. 中国人畜共患病学报, 2009, 25(5):460–464.
[22]潘劲草, 叶榕, 张蔚, 等. 1994年~2004年杭州地区O139群霍乱弧菌携SXT元件和整合子的特征[J]. 中国抗生素杂志, 2007, 32(10):631–635.
[23]YuL, ZhouY, WangR, et al. Multiple antibiotic resistance of Vibrio cholerae serogroup O139 in China from 1993 to 2009[J]. PLoS One, 2012, 7(6):e38633.

用户评论 0条

用户名: 密码: 登陆 注册
 
  • 9
  • 3
  •  4 : 页次:0/0页 共0条记录 5条/每页
    关于我们 | 专家风采 | 会员注册 | 继续教育
    地 址:北京市西城区东河沿街69号正弘大厦511室 邮 编:100052
    电话:010-51322302
    版权所有 中华医学会及中华预防医学杂志编辑部
    京ICP备 07035254 号