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Bacteriology

Identification and Characterization of Clostridium difficile Sequence Type 37 Genotype by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

Ruxin Li, Di Xiao, Jing Yang, Suju Sun, Samuel Kaplan, Zhirong Li, Yanan Niu, Cuixin Qiang, Yu Zhai, Xiaoming Wang, Xingzhen Zhao, Baoxin Zhao, Martin Welker, David H. Pincus, Dazhi Jin, Mini Kamboj, Guanghui Zheng, Guojun Zhang, Jianzhong Zhang, Yi-Wei Tang, Jianhong Zhao
Robin Patel, Editor
Ruxin Li
aDepartment of Clinical Microbiology, Second Hospital of Hebei Medical University, Hebei Provincial Center for Clinical Laboratories, Shijiazhuang, Hebei, China
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Di Xiao
bState Key Laboratory of Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China
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Jing Yang
aDepartment of Clinical Microbiology, Second Hospital of Hebei Medical University, Hebei Provincial Center for Clinical Laboratories, Shijiazhuang, Hebei, China
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Suju Sun
cPublic Health College of Hebei Medical University, Shijiazhuang, Hebei, China
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Samuel Kaplan
hDepartment of Laboratory Medicine, Memorial Sloan Kettering Cancer Center, New York, New York, USA
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Zhirong Li
aDepartment of Clinical Microbiology, Second Hospital of Hebei Medical University, Hebei Provincial Center for Clinical Laboratories, Shijiazhuang, Hebei, China
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Yanan Niu
aDepartment of Clinical Microbiology, Second Hospital of Hebei Medical University, Hebei Provincial Center for Clinical Laboratories, Shijiazhuang, Hebei, China
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Cuixin Qiang
aDepartment of Clinical Microbiology, Second Hospital of Hebei Medical University, Hebei Provincial Center for Clinical Laboratories, Shijiazhuang, Hebei, China
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Yu Zhai
aDepartment of Clinical Microbiology, Second Hospital of Hebei Medical University, Hebei Provincial Center for Clinical Laboratories, Shijiazhuang, Hebei, China
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Xiaoming Wang
aDepartment of Clinical Microbiology, Second Hospital of Hebei Medical University, Hebei Provincial Center for Clinical Laboratories, Shijiazhuang, Hebei, China
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Xingzhen Zhao
aDepartment of Clinical Microbiology, Second Hospital of Hebei Medical University, Hebei Provincial Center for Clinical Laboratories, Shijiazhuang, Hebei, China
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Baoxin Zhao
aDepartment of Clinical Microbiology, Second Hospital of Hebei Medical University, Hebei Provincial Center for Clinical Laboratories, Shijiazhuang, Hebei, China
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Martin Welker
dbioMérieux SA, Marcy-l'Étoile, France
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David H. Pincus
ebioMérieux, Inc., Hazelwood, Missouri, USA
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Dazhi Jin
fDepartment of Microbiology, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou, Zhejiang, China
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  • ORCID record for Dazhi Jin
Mini Kamboj
hDepartment of Laboratory Medicine, Memorial Sloan Kettering Cancer Center, New York, New York, USA
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Guanghui Zheng
gDepartment of Clinical Laboratory, Beijing Tiantan Hospital, Capital Medical University, Beijing, China
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Guojun Zhang
gDepartment of Clinical Laboratory, Beijing Tiantan Hospital, Capital Medical University, Beijing, China
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Jianzhong Zhang
bState Key Laboratory of Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China
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Yi-Wei Tang
hDepartment of Laboratory Medicine, Memorial Sloan Kettering Cancer Center, New York, New York, USA
iDepartment of Pathology and Laboratory Medicine, Weill Medical College of Cornell University, New York, New York, USA
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Jianhong Zhao
aDepartment of Clinical Microbiology, Second Hospital of Hebei Medical University, Hebei Provincial Center for Clinical Laboratories, Shijiazhuang, Hebei, China
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Robin Patel
Mayo Clinic
Roles: Editor
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DOI: 10.1128/JCM.01990-17
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    FIG 1

    Exemplary whole-cell mass spectra of C. difficile strains and isolates of ST37 (ATCC 43598, CD013, CD015, CD017, CD026, and CD126; red lines) and non-ST37 (ATCC BAA-1382, ATCC BAA-1804, ATCC BAA-1870, ATCC BAA-1812, CD105 (ST3), and CD151 (ST2); blue lines). The lower panel (b) shows an enlarged section of the full upper spectra (a).

  • FIG 2
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    FIG 2

    Cluster analysis of spectra of 49 ST37 isolates of different geographic origins. The dendrogram was computed in SARAMIS with a single-linkage agglomerative algorithm-based relative spectrum similarity (percentage of matching peaks within a tolerance of 0.08%). The dotted purple line indicates 65% similarity, which is generally the threshold for intraspecies similarity. Yellow, Hebei; green, Hangzhou; blue, New York; black, ATCC 43598.

Tables

  • Figures
  • TABLE 1

    Sources and sequence types of clinical isolates recruited in this study

    Geographic originSequence typeTotal no. of isolates
    Hebei, China3731
    329
    5423
    214
    3513
    85
    265
    485
    2055
    Other STs48
    Hangzhou, China375
    Hong Kong, China372
    New York City, USA378
    36
    South Korea372
    Singapore371
    351
    Australia21
  • TABLE 2

    Experimental variability of peak height values in spectra of ST37 and non-ST37 isolatesa

    Exptm/zMean peak ht value (integrated in thousands) ± SD for following isolate of ST37 genotype:Mean ± SDCV (%)Mean peak ht value (integrated in thousands) ± SD for following isolate of non-ST37 genotype:Mean ± SDCV (%)ST37/non-ST37 ratioMann-Whitney UP
    CD001CD038CD090CD003CD045CD086
    13,2421,219 ± 1,0355,881 ± 5,4831,516 ± 1,7222,871 ± 3,89913625 ± 4915 ± 268 ± 1416 ± 3220017951<0.001
    3,2866,325 ± 6,15119,169 ± 16,8455,854 ± 6,13110,449 ± 12,249117452 ± 86100151 ± 5253486917<0.001
    3,37014 ± 36142 ± 203280 ± 293145 ± 2271575 ± 970 ± 9941 ± 6431 ± 5618152960.193
    23,242941 ± 1,1771,370 ± 3,142832 ± 8001,048 ± 1,92818411 ± 105 ± 77 ± 148 ± 1113813116<0.001
    3,2864,076 ± 5,0845,082 ± 10,5443,548 ± 3,2154,236 ± 6,7651600000NANA<0.001<0.001
    3,37054 ± 8610 ± 1920 ± 3131 ± 561819 ± 124 ± 729 ± 2813 ± 2015422870.305
    33,242659 ± 6939,448 ± 1,076204 ± 1443,437 ± 7,17320939 ± 5026 ± 2018 ± 2828 ± 331181231<0.001
    3,2861,266 ± 2,11117,937 ± 15,370459 ± 2886,554 ± 11,6821780000NANA<0.001<0.001
    3,370197 ± 10563 ± 12717 ± 2393 ± 11812724 ± 296 ± 717 ± 2516 ± 221386500.182
    • ↵a For experimental conditions and the no. of replicates, see the text. NA, not applicable; SD, standard deviation; CV, coefficient of variation.

  • TABLE 3

    Molecular characteristics of ST37 and non-ST37 C. difficile isolates

    GenotypeGeographic originNo. of isolates testedMSa peak (no. present [%]) at m/z:tcdA or tcdB phenotypeb
    3,2423,2863,370A+B+A−B+A−B−
    ST37Hebei, China3131 (100)30 (96.8)30 (96.8)0 (0)31 (100)0 (0)
    Hangzhou, China55 (100)4 (80.0)5 (100)0 (0)5 (100)0 (0)
    Asia Pacific region55 (100)5 (100)5 (100)0 (0)5 (100)0 (0)
    New York, USA88 (100)8 (100)1 (12.5)NDNDND
    All ST37 origins4949 (100)47 (95.9)41 (83.7)
    Non-ST37Hebei, China14762 (42.2)1 (0.7)135 (91.8)116 (78.9)1 (0.7)30 (20.4)
    Asia Pacific region20 (0)0 (0)2 (100)2 (100)0 (0)0 (0)
    New York, USA62 (33.3)1 (16.7)0 (0)NDNDND
    All non-ST37 origins15564 (41.3)2 (1.3)137 (88.4)
    • ↵a MS, mass spectrometry.

    • ↵b ND, not done.

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Identification and Characterization of Clostridium difficile Sequence Type 37 Genotype by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry
Ruxin Li, Di Xiao, Jing Yang, Suju Sun, Samuel Kaplan, Zhirong Li, Yanan Niu, Cuixin Qiang, Yu Zhai, Xiaoming Wang, Xingzhen Zhao, Baoxin Zhao, Martin Welker, David H. Pincus, Dazhi Jin, Mini Kamboj, Guanghui Zheng, Guojun Zhang, Jianzhong Zhang, Yi-Wei Tang, Jianhong Zhao
Journal of Clinical Microbiology Apr 2018, 56 (5) e01990-17; DOI: 10.1128/JCM.01990-17

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Identification and Characterization of Clostridium difficile Sequence Type 37 Genotype by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry
Ruxin Li, Di Xiao, Jing Yang, Suju Sun, Samuel Kaplan, Zhirong Li, Yanan Niu, Cuixin Qiang, Yu Zhai, Xiaoming Wang, Xingzhen Zhao, Baoxin Zhao, Martin Welker, David H. Pincus, Dazhi Jin, Mini Kamboj, Guanghui Zheng, Guojun Zhang, Jianzhong Zhang, Yi-Wei Tang, Jianhong Zhao
Journal of Clinical Microbiology Apr 2018, 56 (5) e01990-17; DOI: 10.1128/JCM.01990-17
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    • ABSTRACT
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    • MATERIALS AND METHODS
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KEYWORDS

Clostridium difficile
genotype ST37
MALDI-TOF MS
m/z 3,242
m/z 3,286

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