一株mcr-1阳性多重耐药宋内志贺菌的鉴定和药物敏感性特征分析

董念 向莹 胡晓丰 田赛 杜昕颖 邱少富 宋宏彬 孙岩松

引用本文: 董念, 向莹, 胡晓丰, 田赛, 杜昕颖, 邱少富, 宋宏彬, 孙岩松. 一株mcr-1阳性多重耐药宋内志贺菌的鉴定和药物敏感性特征分析[J]. 疾病监测. DOI: 10.3784/j.issn.1003-9961.2019.04.017 shu
Citation:  Nian Dong, Ying Xiang, Xiaofeng Hu, Sai Tian, Xinying Du, Shaofu Qiu, Hongbin Song and Yansong Sun. Identification and antimicrobial resistance of a mcr-1-positive multidrug-resistant Shigella sonnei isolate[J]. Disease Surveillance. DOI: 10.3784/j.issn.1003-9961.2019.04.017 shu

一株mcr-1阳性多重耐药宋内志贺菌的鉴定和药物敏感性特征分析

    作者简介: 董念,男,云南省大理市人,硕士在读,主治医师,主要研究方向为军事预防医学,Email: windy_2005@163.com;
    通信作者: 邱少富, qiushf0613@hotmail.com 宋宏彬, hongbinsong@263.net 孙岩松, sunys1964@hotmail.com
  • 基金项目: 国家重点研发计划 (No. 2017YFC1600105);国家自然科学基金(No. 81872678)

摘要: 目的对分离的1株mcr-1阳性多重耐药宋内志贺菌进行鉴定和耐药性分析。方法采用生化和血清凝集试验进行病原体鉴定;使用微量肉汤稀释法进行药敏试验,并用聚合酶链式反应方法检测耐药基因的存在情况;利用脉冲场凝胶电泳方法对菌株进行遗传多态性和质粒图谱分析并进行Southern杂交试验;用质粒接合转移试验验证mcr-1基因的水平转移能力。结果在广西壮族自治区分离出1株mcr-1阳性的多重耐药宋内志贺菌,对萘啶酸、阿奇霉素、多粘菌素有较高耐药性。 mcr-1基因、mph(A)和blaCTX-M-14基因分别在两个不同的质粒上,且mcr-1基因能通过质粒介导水平转移。结论加强志贺菌尤其是宋内志贺菌对头孢曲松和阿奇霉素共同耐药的监测及mcr-1传播的分子流行病学研究,为合理使用抗生素治疗细菌性痢疾提供依据。

English

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  • 图 1  脉冲场凝胶电泳聚类分析结果

    Figure 1.  PFGE cluster analysis

    图 2  质粒图谱和Southern杂交定位结果

    Figure 2.  Plasmid profiles and Southern hybridization

    图 3  mcr-1基因的PCR扩增结果

    Figure 3.  PCR amplification of mcr-1 gene

    表 1  PCR扩增引物序列

    Table 1.  Primers used for PCR

    耐药基因 引物序列(5′ ~ 3′) 预测长度(bp)
    β-内酰胺类
    blaCTX-M-1 group F:GGTTAAAAAATCACTGCGTC
    R:TTACAAACCGTCGGTGACGA
    873
    blaCTX-M-9 group F:AGAGTGCAACGGATGATG
    R:CCAGTTACAGCCCTTCGG
    868
    blaCTX-M-2/8/25 group F:ACCGAGCCSACGCTCAA
    R:CCGCTGCCGGTTTTATC
    221
    blaTEM F:ATGAGTATTCAACATTTCCG
    R:CCAATGCTTAATCAGTGAGG
    1 080
    blaOXA F:ATTAAGCCCTTTACCAAACCA
    R:AAGGGTTGGGCGATTTTGCCA
    890
    blaVIM F:AGTGGTGAGTATCCGACAG
    R:ATGAAAGTGCGTGGAGAC
    509
    喹诺酮
    qnrA F:ATTTCTCACGCCAGGATTTG
    R:GATCGGCAAAGGTYAGGTCA
    516
    qnrB F:GATCGTGAAAGCCAGAAAGG
    R:ACGAYGCCTGGTAGTTGTCC
    469
    qnrD F:CGAGATCAATTTACGGGGAATA
    R:AACAAGCTGAAGCGCCTG
    656
    qnrS F:ACGACATTCGTCAACTGCAA
    R:TAAATTGGCACCCTGTAGGC
    417
    拓扑异构酶
    gyrA F:TACACCGGTCAACATTGAGG
    R:TTAATGATTGCCGCCGTCGG
    648
    parC F:GTACGTGATCATGGACCGTG
    R:TTCGGCTGGTCGATTAATGC
    531
    大环内酯类耐药
    mph(A) F:CGAAGACCTCCGAGTCCTGC
    R:CCGCCGATACCTCCCAACT
    403
    多粘菌素耐药
    mcr-1 F:CGGTCAGTCCGTTTGTTC
    R:CTTGGTCGGTCTGTAGGG
    291
    下载: 导出CSV

    表 2  宋内志贺菌(Ss2401)药敏试验结果

    Table 2.  Drug susceptibility test of S. sonnei strain (Ss2401)

    抗生素 Ss2401 受体菌J53 J53阳性接合子
    头孢他啶 ≤1(S) ≤1(S) ≤1(S)
    头孢曲松 32(R) ≤1(S) ≤1(S)
    亚胺培南 <4(S) <4(S) <4(S)
    呋喃妥因 ≤32(S) ≤32(S) ≤32(S)
    哌拉西林 >64(R) ≤16(S) ≤16(S)
    四环素 >8(R) ≤4(S) ≤4(S)
    头孢吡肟 ≤8(S) ≤8(S) ≤8(S)
    头孢哌酮 >32(R) ≤16(S) ≤16(S)
    头孢唑啉 >16(R) ≤8(S) ≤8(S)
    头孢西丁 ≤8(S) ≤8(S) ≤8(S)
    妥布霉素 8(I) ≤4(S) ≤4(S)
    左氟沙星 ≤2(S) ≤2(S) ≤2(S)
    庆大霉素 >8(R) ≤4(S) ≤4(S)
    替卡西林 >64(R) ≤16(S) ≤16(S)
    替卡西林/克拉维酸 64(I) ≤16(S) ≤16(S)
    氨曲南 2(S) ≤1(S) ≤1(S)
    氨苄西林 >16(R) ≤8(S) ≤8(S)
    氯霉素 ≤8(S) ≤8(S) ≤8(S)
    复方新诺明 >2(R) ≤2(S) ≤2(S)
    诺氟沙星 ≤4(S) ≤4(S) ≤4(S)
    阿米卡星 ≤16(S) ≤16(S) ≤16(S)
    萘啶酸 >32(R) ≤4(S) ≤4(S)
    阿奇霉素 >128(R) ≤4(S) ≤4(S)
    多粘菌素 >16(R) ≤1(S) >8(R)
     注:表内数据为最低抑制浓度(μg/ml); R. 耐药;I. 中等;S. 敏感
    下载: 导出CSV
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