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浙江地区耐多药结核分枝杆菌基因突变特征分析

赵刚 贾庆军 吴亦斐 黄银燕 唐磊明 曾美纯

赵刚, 贾庆军, 吴亦斐, 黄银燕, 唐磊明, 曾美纯. 浙江地区耐多药结核分枝杆菌基因突变特征分析[J]. 中华疾病控制杂志, 2021, 25(1): 66-71. doi: 10.16462/j.cnki.zhjbkz.2021.01.013
引用本文: 赵刚, 贾庆军, 吴亦斐, 黄银燕, 唐磊明, 曾美纯. 浙江地区耐多药结核分枝杆菌基因突变特征分析[J]. 中华疾病控制杂志, 2021, 25(1): 66-71. doi: 10.16462/j.cnki.zhjbkz.2021.01.013
ZHAO Gang, JIA Qing-jun, WU Yi-fei, HUANG Yin-yan, TANG Lei-ming, ZENG Mei-chun. Analysis for characteristics of genetic mutations among multi-drug resistant Mycobacterium tuberculosis (MDR-TB) isolates from Zhejiang Province[J]. CHINESE JOURNAL OF DISEASE CONTROL & PREVENTION, 2021, 25(1): 66-71. doi: 10.16462/j.cnki.zhjbkz.2021.01.013
Citation: ZHAO Gang, JIA Qing-jun, WU Yi-fei, HUANG Yin-yan, TANG Lei-ming, ZENG Mei-chun. Analysis for characteristics of genetic mutations among multi-drug resistant Mycobacterium tuberculosis (MDR-TB) isolates from Zhejiang Province[J]. CHINESE JOURNAL OF DISEASE CONTROL & PREVENTION, 2021, 25(1): 66-71. doi: 10.16462/j.cnki.zhjbkz.2021.01.013

浙江地区耐多药结核分枝杆菌基因突变特征分析

doi: 10.16462/j.cnki.zhjbkz.2021.01.013
基金项目: 

杭州市科技发展计划项目 20191203B140

杭州市卫生健康科技计划项目 A20200788

详细信息
    通讯作者:

    贾庆军,E-mail: qingjunjia@126.com

  • 中图分类号: R52

Analysis for characteristics of genetic mutations among multi-drug resistant Mycobacterium tuberculosis (MDR-TB) isolates from Zhejiang Province

Funds: 

Hangzhou Science and Technology Development Plan Project 20191203B140

Hangzhou Health Science and Technology Plan Project A20200788

More Information
  • 摘要:   目的  分析浙江地区耐多药结核分枝杆菌(M.tuberculosis complex, MTBC)异烟肼(isoniazid, INH)和利福平(rifampicin, RIF)耐药相关基因突变特征。  方法  对患者痰标本阳性培养物进行菌种鉴定,提取耐多药菌株DNA,应用PCR法对inhAkatGrpoB RRDR区进行扩增,并将扩增产物直接进行基因测序。  结果  共鉴定出3 317株MTBC,对应的患者中初治为3 009例,复治为308例,耐多药结核病(multidrug-resistant tuberculosis, MDR-TB)为84例,占比为2.53%(84/3 317)。其中,在INH耐药方面,47株分别在katG(315)或inhA(-15/-8)发生单位点突变,19株同时出现katG(315)和inhA(-15/-8)突变,18株未检测出上述基因突变,基因型同表型的符合率为78.57%(66/84),突变频率最高的位点是Ser315Thr,占47.62%(40/84)。在RIF耐药方面,79株在rpoB(511、512、513、516、522、526和531位点)出现突变,2株同时出现511/513和511/516突变,突变频率最高的位点是Ser531Leu,占58.33%(49/84),3株未检测出rpoB突变,基因型同表型的符合率为96.43%(81/84)。  结论  浙江地区MDR-TB流行株基因突变仍然集中在常见的基因位点,表明在本地区利用Xpert MTB/RIF筛查MDR-TB患者仍具有较高的应用价值。
  • 图  1  研究流程简图

    Figure  1.  Study flowchart

    图  2  基因扩增产物电泳

    注:泳道1~4分别为katGinhArpoB和100 bp DNA marker。

    Figure  2.  Electrophoresis of gene amplification products

    表  1  84例耐多药患者特征分析

    Table  1.   Analysis for characteristics of 84 MDR-TB patients

    特征 例数 占比(%)
    性别
      男 56 66.67
      女 28 33.33
    年龄(岁)
      ≤30 25 29.76
      30~ 21 25.00
      ≥50 38 45.24
    户籍
      本地 61 72.62
      外地 23 27.38
    职业
      体力劳动者 55 65.48
      非体力劳动者 29 34.52
    实验室检测
      抗酸染色阳性 57 67.86
    治疗结果
      治愈或完成疗程 65 77.38
      治疗中 11 13.10
      失败 8 9.52
    肺结核分类
      原发性肺结核 12 14.29
      继发性肺结核 72 85.71
    下载: 导出CSV

    表  2  84株表型耐药及10株表型敏感MTBC katGinhA区突变位点分布情况

    Table  2.   Distributions of genetic mutations of katG and inhA among 84 INHR and 10 INHSa

    株型 突变类型 位点 核苷酸和(或)氨基酸改变 株数(N=94)
    核苷酸改变 氨基酸改变
    耐药株 未检测出 18
    单位点突变 katG 315 AGC→ACC Ser→Thr 21
    AGC→TGC Ser→Cys 8
    AGC→AAC Ser→Asn 5
    inhA -15 C→T 9
    inhA -8 T→C 4
    双位点突变 katG 315/inhA -15 AGC→ACC Ser→Thr 13
    C→T
    katG 315/inhA -8 AGC→ACC Ser→Thr 6
    T→C
    敏感株 未检出 10
    注:aINHR表示异烟肼耐药株,INHS表示异烟肼敏感株。
    下载: 导出CSV

    表  3  84株表型耐药及10株表型敏感MTBC rpoB核心区突变位点分布情况

    Table  3.   Distributions of genetic mutations of rpoB among 84 RIFR and 10 RIFSa

    株型 突变类型 位点 核苷酸和(或)氨基酸改变 株数(N=94)
    核苷酸改变 氨基酸改变
    耐药株 未检测出 3
    单位点突变 511 CTG→CCG Leu→Pro 1
    512 AGC→ACC Ser→Thr 2
    513 CAA→AAA Gln→Lys 1
    516 GAC→GTC Asp→Val 3
    522 TCG→TTG Ser→Leu 2
    526 CAC→GAC His→Asp 13
    CAC→CCC His→Pro 2
    CAC→CTC His→Leu 1
    CAC→CGC His→Arg 2
    CAC→AAC His→Asn 1
    531 TCG→TTG Ser→Leu 49
    TCG→TGG Ser→Trp 2
    双位点突变 511/513 CTG→CCG Leu→Pro 1
    CAA→AAA Gln→Lys
    511/516 CTG→CCG Leu→Pro 1
    GAC→GTC Asp→Val
    敏感株 未检出 10
    注:aINHR表示异烟肼耐药株,INHS表示异烟肼敏感株。
    下载: 导出CSV
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  • 收稿日期:  2019-12-24
  • 修回日期:  2020-04-20
  • 刊出日期:  2021-01-10

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