Copy Number Change of the NDM-1 Sequence in a Multidrug-Resistant Klebsiella pneumoniae Clinical Isolate

Tzu Wen Huang, Te Li Chen, Ying Tsong Chen, Tsai Ling Lauderdale, Tsai Lien Liao, Yi Tzu Lee, Chien Pei Chen, Yen Ming Liu, Ann Chi Lin, Ya Hui Chang, Keh Ming Wu, Ralph Kirby, Jui Fen Lai, Mei Chen Tan, Leung Kei Siu, Chung Ming Chang, Chang Phone Fung, Shih Feng Tsai

Research output: Contribution to journalArticle

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Abstract

The genetic features of the antimicrobial resistance of a multidrug resistant Klebsiella pneumoniae strain harboring blaNDM-1 were investigated to increase our understanding of the evolution of NDM-1. The strain, KPX, came from a Taiwanese patient with a hospitalization history in New Delhi. Complete DNA sequencing was performed; and the genes responsible for antimicrobial resistance were systematically examined and isolated by library screening. KPX harbored two resistance plasmids, pKPX-1 and pKPX-2, which are 250-kb and 141-kb in size, respectively, with blaNDM-1 present on pKPX-1. The plasmid pKPX-1 contained genes associated with the IncR and IncF groups, while pKPX-2 belonged to the IncF family. Each plasmid carried multiple antimicrobial resistance genetic determinants. The gene responsible for resistance to carbapenems was found on pKPX-1 and that for resistance to aztreonam was found on pKPX-2. To our surprise, we discovered that blaNDM-1 exists on pKPX-1 as multiple copies in the form of tandem repeats. Amplification of blaNDM-1 was found to occur by duplication of an 8.6-kb unit, with the copy number of the repeat varying from colony to colony. This repeat sequence is identical to that of the pNDM-MAR except for two base substitutions. The copy number of blaNDM-1 of colonies under different conditions was assessed by Southern blotting and quantitative PCR. The blaNDM-1 sequence was maintained in the presence of the antimicrobial selection; however, removal of antimicrobial selection led to the emergence of susceptible bacterial populations with a reduced copy number or even the complete loss of the blaNDM-1 sequence. The dynamic nature of the NDM-1 sequence provides a strong argument for judicious use of the broad-spectrum antimicrobials in order to reduce the development and spread of antimicrobial resistance among pathogens.

Original languageEnglish
Article numbere62774
JournalPLoS One
Volume8
Issue number4
DOIs
Publication statusPublished - Apr 29 2013
Externally publishedYes

Fingerprint

Klebsiella pneumoniae
antibiotic resistance
Plasmids
Genes
plasmids
anti-infective agents
Aztreonam
Carbapenems
Tandem Repeat Sequences
aztreonam
carbapenems
Multiple Drug Resistance
Pathogens
Southern Blotting
DNA Sequence Analysis
Amplification
tandem repeat sequences
genes
Screening
Hospitalization

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Huang, T. W., Chen, T. L., Chen, Y. T., Lauderdale, T. L., Liao, T. L., Lee, Y. T., ... Tsai, S. F. (2013). Copy Number Change of the NDM-1 Sequence in a Multidrug-Resistant Klebsiella pneumoniae Clinical Isolate. PLoS One, 8(4), [e62774]. https://doi.org/10.1371/journal.pone.0062774

Copy Number Change of the NDM-1 Sequence in a Multidrug-Resistant Klebsiella pneumoniae Clinical Isolate. / Huang, Tzu Wen; Chen, Te Li; Chen, Ying Tsong; Lauderdale, Tsai Ling; Liao, Tsai Lien; Lee, Yi Tzu; Chen, Chien Pei; Liu, Yen Ming; Lin, Ann Chi; Chang, Ya Hui; Wu, Keh Ming; Kirby, Ralph; Lai, Jui Fen; Tan, Mei Chen; Siu, Leung Kei; Chang, Chung Ming; Fung, Chang Phone; Tsai, Shih Feng.

In: PLoS One, Vol. 8, No. 4, e62774, 29.04.2013.

Research output: Contribution to journalArticle

Huang, TW, Chen, TL, Chen, YT, Lauderdale, TL, Liao, TL, Lee, YT, Chen, CP, Liu, YM, Lin, AC, Chang, YH, Wu, KM, Kirby, R, Lai, JF, Tan, MC, Siu, LK, Chang, CM, Fung, CP & Tsai, SF 2013, 'Copy Number Change of the NDM-1 Sequence in a Multidrug-Resistant Klebsiella pneumoniae Clinical Isolate', PLoS One, vol. 8, no. 4, e62774. https://doi.org/10.1371/journal.pone.0062774
Huang, Tzu Wen ; Chen, Te Li ; Chen, Ying Tsong ; Lauderdale, Tsai Ling ; Liao, Tsai Lien ; Lee, Yi Tzu ; Chen, Chien Pei ; Liu, Yen Ming ; Lin, Ann Chi ; Chang, Ya Hui ; Wu, Keh Ming ; Kirby, Ralph ; Lai, Jui Fen ; Tan, Mei Chen ; Siu, Leung Kei ; Chang, Chung Ming ; Fung, Chang Phone ; Tsai, Shih Feng. / Copy Number Change of the NDM-1 Sequence in a Multidrug-Resistant Klebsiella pneumoniae Clinical Isolate. In: PLoS One. 2013 ; Vol. 8, No. 4.
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