深圳地区致泻性大肠埃希菌多位点可变数目串联重复序列分析分型方法的研究

王磊 陈琼城 胡璐璐 邱亚群 林一曼 江敏 姜伊祥 石晓路 扈庆华 李迎慧

王磊, 陈琼城, 胡璐璐, 邱亚群, 林一曼, 江敏, 姜伊祥, 石晓路, 扈庆华, 李迎慧. 深圳地区致泻性大肠埃希菌多位点可变数目串联重复序列分析分型方法的研究[J]. 疾病监测, 2019, 34(10): 905-911. doi: 10.3784/j.issn.1003-9961.2019.10.010
引用本文: 王磊, 陈琼城, 胡璐璐, 邱亚群, 林一曼, 江敏, 姜伊祥, 石晓路, 扈庆华, 李迎慧. 深圳地区致泻性大肠埃希菌多位点可变数目串联重复序列分析分型方法的研究[J]. 疾病监测, 2019, 34(10): 905-911. doi: 10.3784/j.issn.1003-9961.2019.10.010
Lei Wang, Qiongcheng Chen, Lulu Hu, Yaqun Qiu, Yiman Lin, Min Jiang, Yixiang Jiang, Xiaolu Shi, Qinghua Hu, Yinghui Li. Evaluation of multiple locus variable-number tandem repeat analysis for subtyping of diarrheagenic Escherichia coli in Shenzhen[J]. Disease Surveillance, 2019, 34(10): 905-911. doi: 10.3784/j.issn.1003-9961.2019.10.010
Citation: Lei Wang, Qiongcheng Chen, Lulu Hu, Yaqun Qiu, Yiman Lin, Min Jiang, Yixiang Jiang, Xiaolu Shi, Qinghua Hu, Yinghui Li. Evaluation of multiple locus variable-number tandem repeat analysis for subtyping of diarrheagenic Escherichia coli in Shenzhen[J]. Disease Surveillance, 2019, 34(10): 905-911. doi: 10.3784/j.issn.1003-9961.2019.10.010

深圳地区致泻性大肠埃希菌多位点可变数目串联重复序列分析分型方法的研究

doi: 10.3784/j.issn.1003-9961.2019.10.010
基金项目: 2013年度深圳市科技计划项目(No.201302142);深圳市医疗三名工程(No.SZSM20181107)
详细信息
    作者简介:

    王磊,男,河南省新乡市人,硕士,研究实习员,主要从事疑难菌的鉴定、细菌耐药和病原菌特征研究,Email:wangl@szcdc.net

    通讯作者:

    李迎慧,Tel:0755–25537785,Email:liyh@szcdc.net

  • 中图分类号: R378.2+1

Evaluation of multiple locus variable-number tandem repeat analysis for subtyping of diarrheagenic Escherichia coli in Shenzhen

Funds: This study was supported by the found of Shenzhen Science and Technology Project in 2013 (No.201302142); and Shenzhen's Sanming Project (No.SZSM20181107)
More Information
  • 摘要: 目的通过比较多位点可变数目串联重复序列分析(MLVA)和脉冲场凝胶电泳(PFGE)分型方法,探索更优的致泻性大肠埃希菌(DEC)的快速溯源方法。方法本研究采用筛选的可变数目串联重复序列(VNTR)位点,对DEC菌株进行MLVA分型,并将该方法与金标准PFGE进行比较评估。结果PFGE分型方法将58株DEC分为43种带型,其中3种带型成簇,多样性指数(DI)值为0.965。 MLVA分型方法将58株菌分为42种带型,4种型别成簇,DI值为0.955。 此外,2种方法对2起暴发菌株的分型结果一致。结论MLVA方法与PFGE方法对深圳地区的58株DEC菌株的分型能力差异无统计学意义,但MLVA具有快速、简便、通量高的特点,使得MLVA优于PFGE分型方法,在疾病监测、疾病暴发调查中将会发挥重要的应用价值。
  • 图  1  58株致泻性大肠埃希菌的PFGE分型和聚类分析

    注:ETEC. 肠产毒性大肠埃希菌;EPEC. 肠致病性大肠埃希菌;EIEC. 肠侵袭性大肠埃希菌;EAEC. 肠聚集性大肠埃希菌;STEC. 产志贺毒素大肠埃希菌;OUT. 与52种O抗原抗血清均不凝集;HNT. 与22种H抗原抗血清均不凝集;HNM. 无动力

    Figure  1.  PFGE patterns and cluster analysis results of 58 diarrheagenic E. coli strains

    图  2  MLVA与PFGE两种分型方法的聚类结果比较

    注:方框内菌株为与PFGE分型有差异的菌株,圆圈内表示CVN014位点有差异

    Figure  2.  Comparison of clustering results of M LVA and PFGE

    图  3  2起暴发疫情菌株的MLVA与PFGE聚类结果

    注:A、B各代表一起暴发事件;ADEC. 非典型致泻性大肠埃希菌;EPEC. 肠致病性大肠埃希菌

    Figure  3.  Comparison of MLVA and PFGE clustering results of two outbreak strains

    表  1  实验菌株

    Table  1.   Strains used in the study

    菌株编号致病型别血清型 菌株编号致病型别血清型
    E12194EAECO18E14064ETECO15∶HNT
    E11184EAECO44E11450ETECO159∶H34
    E12117EAECO86aE10219ETECO159∶H7
    E10113EAECOUTE10060ETECO159∶HNT
    E13141EAECOUTE11405ETECO169∶H41
    E13096EAECOUTE14083ETECO25∶H4
    E14011EAECOUTE11454ETECO25∶H12
    E12251EAECOUTE10178ETECO27∶H7
    E13035EAECOUTE09065ETECO27∶H7
    E11588EIECO1E11439ETECO27∶HNT
    E11564EIECO153E09021EIECO27∶HNT
    E10200EIECO28acE11345ETECO6∶H16
    E10118EIECO28acE14078ETECO6∶HNM
    E14068EPECO111E14045ETECO148∶H28
    E14042EPECO128E14049ETECO148∶H28
    E14053EPECO145E15006ETECO148∶H28
    E10387EPECO153E11116ETECO148∶H28
    E11178EPECO166E11325ETECO148∶H28
    E11167EPECO18E10345ETECO148∶HNT
    E13063EPECO25E11452ETECO148∶H28
    E13095EPECO26E11266ETECO148∶H28
    E11537EPECO27E11267ETECO148∶H28
    E10359EPECO29E11299ETECO148∶H28
    E11296EPECOUTE11346ETECO148∶H28
    E14030EPECOUTE11363ETECO148∶H28
    E10341STECO1E11424ETECO148∶H28
    E11008ETECO159∶H34E11449ETECO148∶H28
    E14047ETECO159∶H34E10361ETECO148∶HNT
    E11385ETECO115∶HNTE10403ETECO148∶HNT
    E13015ADECO112ac∶H7E13020ADECO112ac∶H7
    E13016ADECO112ac∶H7E10065EPECO29
    E13017ADECO112ac∶H7E10066EPECO29
    E13019ADECO112ac∶H7E10067EPECO29
      注:ETEC. 肠产毒性大肠埃希菌;EPEC. 肠致病性大肠埃希菌;EIEC. 肠侵袭性大肠埃希菌;EAEC. 肠聚集性大肠埃希菌;STEC. 产志贺毒素大肠埃希菌;ADEC. 非典型致泻性大肠埃希菌;OUT. 与52种O抗原抗血清均不凝集;HNT. 与22种H抗原抗血清均不凝集;HNM. 无动力
    下载: 导出CSV

    表  2  实验采用的7个VNTR位点的序列和PCR扩增引物

    Table  2.   Sequences of the 7 VNTR loci and primers for PCR amplification used in the study

    VNTR位点序列长度(bp)引物名称引物序列(5′ ~ 3′)
    CVN001CAGCAGCCGCAACAACCGGTT39001FAACCGGCTGGGGCGAATCC
    GCGCCGCAGCAGCAATAT001RGGCGGCGGTGTCAGCAAATC
    CVN002TTAAATAATTCACAGGAG18002FAACCGTTATGAARGRAAGTCCT
    002RTCGCCCAGTAAGTATGAAATC
    CVN003TGCTACCCTGGACGG15003FAAAAATCCGGATGAGWTGGTC
    003RTTGCGTTGTCAGTAATTTGTTCAG
    CVN004AAAGCAGCAGCTGAA15004FMGCTGCGGCRCTGAAGAAGA
    004RCCCGGCAGGCGAAGCATTGT
    CVN007CATCATCACGATCACGAA18007FACCGTGGCTCCAGYTGATTTC
    007RACCAGTGTTGCGCCCAGTGTC
    CVN014TGCAGG 6014FTCCCCGCAATCAGCAAMACAAAGA
    014RGCAGCRGGGACAACGGAAGC
    CVN015CATTACCACCAC12015FTAGGCATAGCGCACAGACAGATAA
    015RGTACCGCCGAACTTCAACACTC
    下载: 导出CSV
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  • 收稿日期:  2019-03-29
  • 网络出版日期:  2019-10-08
  • 刊出日期:  2019-10-01

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