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blaNDM-5-Bearing IncFII-Type Plasmids of Klebsiella pneumoniae Sequence Type 147 Transmitted by Cross-Border Transfer of a Patient Juyoun Shin,a Jin Yang Baek,b Sun Young Cho,d Hee Jae Huh,c Nam Yong Lee,c Jae-Hoon Song,b,d Doo Ryeon Chung,b,d Kwan Soo Koa,b Department of Molecular Cell Biology, Sungkyunkwan University School of Medicine, Suwon, South Koreaa; Asia Pacific Foundation for Infectious Diseases (APFID), Seoul, South Koreab; Department of Laboratory Medicine, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, South Koreac; Division of Infectious Diseases, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, South Koread
The two plasmids extracted from Klebsiella pneumoniae sequence type 147 (ST147) isolates were analyzed. The first isolate was obtained from a patient transferred from United Arab Emirates to South Korea. The second isolate was obtained from a Korean patient and was suspected to be transmitted from the first patient. Sequences of two plasmids were almost the same, and genetic structures, including blaNDM-5, of these plasmids were similar to plasmids of NDM-1-producing Escherichia coli ST131 isolates found in Europe.
T
hus, far, 13 variants of NDM have been identified (1). Of these, NDM-5 has been repeatedly isolated, mainly from Escherichia coli found in European, Asian, and African countries (2–7). Recently, two NDM-5-producing Klebsiella pneumoniae isolates (CC1409-1 and CC1410-1) were identified in South Korea (8). The first isolate was collected from a patient who was transferred from Abu Dhabi, United Arab Emirates (UAE), to Seoul, South Korea, and the second isolate was collected from a Korean patient after 4 months. Because both the isolates belonged to the same clone and shared many antimicrobial resistance genes, the second isolate was thought to have been transmitted by cross-border transfer from the patient from UAE (8). In the present study, we determined the complete sequences of the two blaNDM-5-bearing plasmids (pCC1409-1 and pCC1410-1) isolated from the K. pneumoniae isolates. We confirmed that the two plasmids had similar sequences, with minor differences, and were very similar to blaNDM-1-bearing IncFII-type plasmids of E. coli isolates found in Europe. These plasmids were conjugated using E. coli J53, a sodium azide-resistant and plasmid-free strain, as a recipient, as in the previous study (9). The MiSeq desktop sequencer system (Illumina, Inc., San Diego, CA) was used to determine the complete sequence of the plasmids pCC1409-1 and pCC1410-1. A total of 3,768,155 and 3,825,823 sequence reads were generated, yielding an average sequencing depth of 11,845⫻ and 13,312⫻ for the plasmids pCC1409-1 and pCC1410-1, respectively. PCR combinations were used to close gaps between the contigs and to confirm the position of the contigs (see the supplemental material). Open reading frames were predicted, annotated using the Glimmer 3.0 system (http://ccb.jhu.edu/software/glimmer /index.shtml), and confirmed using DNAMAN 5.2.10 software (Lynnon BioSoft, Lynnon Corporation). Each predicted protein was compared against an all-protein database by using BLASTP (http://blast.ncbi.nlm.nih.gov/blast.cgi). Gene sequences were compared and aligned with those of genes in pGUE-NDM and pMC-NDM by using cross_match 1.080812 software and ClustalW (http://www.ebi.ac.uk/Tools/msa/clustalw2). Whole-plasmid sequencing indicated that plasmid pCC1409-1 isolated from the K. pneumoniae CC1409-1 isolate collected from the patient transferred from UAE was 89,745 bp in size and had 114 protein coding sequences (CDS). The other plasmid, pCC1410-1, of the K. pneumoniae CC1410-1 isolate col-
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lected from the Korean patient was 95,005 bp in size and was predicted to harbor 120 CDS. Both of the plasmids belonged to the incompatibility group IncFII, with their G⫹C contents being 53.04% and 52.96%, respectively. Comparison of their genetic structures indicated that these plasmids carried almost the same genes, except that pCC1410-1 had an ISCR3 insertion sequence containing erm genes that encoded rRNA adenine N-6-methyltransferase (Fig. 1). The strain isolated from the first patient may have been transmitted to the second patient through unknown transmitters, hospital equipment, hospital staffs, or other patients. During the transmission, a small downstream region of blaNDM-5 may have been incorporated into the second plasmid, pCC1410-1. Comparison of databases indicated that pCC1409-1 and pCC1410-1 shared 99% sequence similarity with both pGUENDM (GenBank accession no. JQ364967.1) and pMC-NDM (GenBank accession no. HG003695.1) carrying NDM-1 genes, which were isolated from carbapenem-resistant E. coli ST131 isolates found in France and Poland, respectively (Fig. 1) (10, 11). In particular, backbone structures, including replication, partition, and conjugative transfer loci, of pCC1409-1 and pCC1410-1 showed very high nucleotide similarity with those of pGUE-NDM and pMC-NDM. All of the 24 tra and 8 trb genes, which contributed to plasmid transferability of pCC1409-1 and pCC1410-1, showed 97% and 95% sequence similarity, respectively, to those in pGUE-NDM and pMC-NDM. These transfer loci of pCC1409-1 and pCC1410-1 were more similar to those of blaCTX-M-15-bearing
Received 10 November 2015 Returned for modification 6 December 2015 Accepted 21 December 2015 Accepted manuscript posted online 11 January 2016 Citation Shin J, Baek JY, Cho SY, Huh HJ, Lee NY, Song J-H, Chung DR, Ko KS. 2016. blaNDM-5-bearing IncFII-type plasmids of Klebsiella pneumoniae sequence type 147 transmitted by cross-border transfer of a patient. Antimicrob Agents Chemother 60:1932–1934. doi:10.1128/AAC.02722-15. Address correspondence to Doo Ryeon Chung, [email protected], or Kwan Soo Ko, [email protected]. Supplemental material for this article may be found at http://dx.doi.org/10.1128 /AAC.02722-15. Copyright © 2016, American Society for Microbiology. All Rights Reserved.
Antimicrobial Agents and Chemotherapy
March 2016 Volume 60 Number 3
blaNDM-5-Bearing K. pneumoniae Plasmids
FIG 1 Comparison of the major structural features of pCC1409-1 and pCC1410-1 with those of IncFII-type plasmids pGUE-NDM (GenBank accession no. JQ364967.1) and pMC-NDM (GenBank accession no. HG003695.1). White boxes indicate plasmid scaffold regions that are common among the plasmids. The tra loci are indicated within the white boxes by capital letters. Resistance genes are indicated by gray boxes, and transposon-related genes, integrase, and insertion sequences are indicated by diagonally lined boxes. Replicase genes are indicated by dots, and addiction system genes are indicated in black. Light gray shading denotes regions of homology, and arrows indicate blaNDM-type genes.
pC15-1a (GenBank accession no. AY458016.1) and pHK23a (GenBank accession no. JQ432559.1) isolated from E. coli isolates found in Canada and Hong Kong, respectively (12, 13). In addition, both pCC1409-1 and pCC1410-1 contained identical IncFII replicons repA2-repA6-repA1-repA4, a 2,166-bp fragment (G⫹C content, 51.89%; dotted boxes in Fig. 1), that showed high nucleotide similarity to IncFII replicons of pMCNDM and pGUE-NDM. pCC1409-1 and pCC1410-1 contained 11 and 13 antimicrobial resistance genes, respectively, including a blaNDM-type-bleMBL-trpF-tat module and the -lactamase gene blaTEM-1 (Fig. 1). In addition, genes conferring resistance against macrolides [mph(A), mrx, and mphR(A)], sulfonamides (sul1), quaternary ammonium compounds (qacE⌬1), aminoglycosides (aadA4 and rmtB), and trimethoprim (dfrA17) were identified in the antimicrobial resistance region of pCC1409-1 and pCC1410-1. The upstream and downstream structures of blaNDM-5 were almost identical to those of blaNDM-1 in pMC-NDM (Fig. 1). NDM-5 differs from NDM-1 by two amino acid substitutions at positions 88 (Val¡Leu) and 154 (Met¡Leu) (2). The blaNDM-5bearing plasmids examined in the present study had the same origin as pMC-NDM. However, the proximal part of the resistance region [lqsb]from insAB to mphR(A)] in pCC1409-1 and pCC1410-1 was different from that in pGUE-NDM and pMCNDM (Fig. 1). The adjacent structure of blaNDM-5 has been identified in other NDM-5-producing E. coli isolates. Although complete sequences of plasmids of these E. coli isolates have not been determined, the proximal structure of their resistance region is suggested to be different from that of plasmids examined in the present study (3, 5). Although blaNDM-5-bearing pCC1409-1 and pCC1040-1 had the same origin as blaNDM-1-bearing pMC-NDM, the proximal part of their resistance region might have originated from other plasmids. Further, blaNDM-5-containing region of pCC1409-1 and pCC1410-1 differed from that of a plasmid isolated from NDM1-producing K. pneumoniae isolates found in South Korea (14).
March 2016 Volume 60 Number 3
Both pCC1409-1 and pCC1410-1 were isolated from K. pneumoniae ST147 isolates. K. pneumoniae ST147 isolates that are resistant to diverse antibiotics have disseminated worldwide (15– 19); however, ST147 isolates, to our knowledge, have not been found in K. pneumoniae from South Korea. Therefore, we suggest that pCC1409-1 and pCC1410-1 were not derived from K. pneumoniae isolates found in South Korea. In this study, we determined the complete sequences of the two IncFII-type plasmids isolated from NDM-5-producing K. pneumoniae isolates and observed that the two plasmids had very similar sequences. To our knowledge, this is the first study to report the complete sequences of blaNDM-5-bearing plasmids. Nucleotide sequence accession numbers. The annotated sequences of pCC1409-1 and pCC1410-1 have been submitted to GenBank nucleotide sequence database under accession numbers KT725789 and KT725788, respectively. ACKNOWLEDGMENTS This research was supported by the Basic Science Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT, and Future Planning (grant NRF2013R1A2A2A0104103). J. Shin was partly supported by the Basic Science Research Program through the NRF funded by the Ministry of Education (grant 2013R1A12062884).
REFERENCES 1. Shrestha B, Tada T, Miyoshi-Akiyama T, Shimada K, Ohara H, Kirikae T, Pokhrel BM. 2015. Identification of a novel variant, NDM-13, from a multidrug-resistant Escherichia coli from Nepal. Antimicrob Agents Chemother 59:5847–5850. http://dx.doi.org/10.1128/AAC.00332-15. 2. Hornsey M, Phee L, Wareham DW. 2011. A novel variant, NDM-5, of the New Delhi metallo--lactamase in a multidrug-resistant Escherichia coli ST648 isolate recovered from a patient in the United Kingdom. Antimicrob Agents Chemother 55:5952–5954. http://dx.doi.org/10.1128/AAC .05108-11. 3. Yang P, Xie Y, Feng P, Zong Z. 2014. blaNDM-5 carried by an IncX3 plasmid in Escherichia coli sequence type 167. Antimicrob Agents Chemother 58:7548 –7552. http://dx.doi.org/10.1128/AAC.03911-14.
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4. Nakano R, Nakano A, Hikosaka K, Kawakami S, Matsunaga N, Asahara M, Ishigaki S, Furukawa T, Suzuki M, Shibayama K, Ono Y. 2014. First report of metallo--lactamase NDM-5-producing Escherichia coli in Japan. Antimicrob Agents Chemother 58:7611–7612. http://dx.doi.org/10 .1128/AAC.04265-14. 5. Pitart C, Solé M, Roca I, Román A, Moreno A, Vila J, Marco F. 2015. Molecular characterization of blaNDM-5 carried on an IncFII plasmid in an Escherichia coli isolate from a nontraveler patient in Spain. Antimicrob Agents Chemother 59:659 – 662. http://dx.doi.org/10.1128/AAC .04040-14. 6. Rahman M, Shukla SK, Prasad KN, Ovejero CM, Pati BK, Tripathi A, Singh A, Srivastava AK, Gonzalez-Zorn B. 2014. Prevalence and molecular characterization of New Delhi metallo--lactamases NDM-1, NDM-5, NDM-6, and NDM-7 in multidrug-resistant Enterobacteriaceae from India. Int J Antimicrob Agents 44:30 –37. http://dx.doi.org/10.1016 /j.ijantimicag.2014.03.003. 7. Sassi A, Loucif L, Gupta SK, Dekhil M, Chettibi H, Rolain JM. 2014. NDM-5 carbapenemase-encoding gene in multidrug-resistant clinical isolates of Escherichia coli from Algeria. Antimicrob Agents Chemother 58:5606 –5608. http://dx.doi.org/10.1128/AAC.02818-13. 8. Cho SY, Huh HJ, Baek JY, Chung NY, Ryu JG, Ki CS, Chung DR, Lee NY, Song JH. 2015. Klebsiella pneumoniae coproducing NDM-5 and OXA-181 carbapenemases, South Korea. Emerg Infect Dis 21:1088 –1089. http://dx.doi.org/10.3201/eid2106.150048. 9. Song W, Lee H, Lee K, Jeong SH, Bae IK, Kim JS, Kwak HS. 2009. CTX-M-14 and CTX-M-15 enzymes are the dominant type of extendedspectrum -lactamase in clinical isolates of Escherichia coli from Korea. J Med Microbiol 58:261–266. http://dx.doi.org/10.1099/jmm.0.004507-0. 10. Bonnin RA, Poirel L, Carattoli A, Nordmann P. 2012. Characterization of an IncFII plasmid of an IncFII plasmid encoding NDM-1 from Escherichia coli ST131. PLoS One 7:e34752. http://dx.doi.org/10.1371/journal .pone.0034752. 11. Fiett J, Baraniak A, Izdebski R, Sitkiewicz I, Z˙abicka D, Meler A, Filczak K, Hryniewicz W, Gniadkowski M. 2014. The first NDM metallo-lactamase-producing Enterobacteriaceae isolate in Poland: evolution of IncFII-type plasmids carrying the blaNDM-1 gene. Antimicrob Agents Chemother 58:1203–1207. http://dx.doi.org/10.1128/AAC.01197-13. 12. Boyd DA, Tyler S, Christianson S, McGeer A, Muller MP, Willey BM,
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Bryce E, Gardam M, Nordmann P, Mulvey MR. 2004. Complete nucleotide sequence of a 92-kilobase plasmid harboring the CTX-M-15 extended-spectrum -lactamase involved in an outbreak in long-term-care facilities in Toronto, Canada. Antimicrob Agents Chemother 48:3758 –3764. http://dx.doi.org/10.1128/AAC.48.10.3758-3764.2004. Ho PL, Chan J, Law PY, Law PW, Chow KH. 2013. Plasmid-mediated fosfomycin resistance in Escherichia coli isolated from pig. Vet Microbiol 162:964 –967. http://dx.doi.org/10.1016/j.vetmic.2012.09.023. Kim SY, Rhee JY, Shin SY, Ko KS. 2014. Characteristics of communityonset NDM-1-producing Klebsiella pneumoniae isolates. J Med Microbiol 63:86 – 89. Damjanova I, Tóth A, Pászti J, Hajbel-Vékony G, Jakab M, Berta J, Milch H, Füzi M. 2008. Expansion and countrywide dissemination of ST11, ST15, and ST147 ciprofloxacin-resistant CTX-M-15-type -lactamase-producing Klebsiella pneumoniae epidemic clones in Hungary in 2005: the new MRSAs? J Antimicrob Chemother 62:978 –985. http://dx .doi.org/10.1093/jac/dkn287. Wang G, Huang T, Surendraiah PK, Wang K, Komal R, Zhuge J, Chern CR, Kryszuk AA, King C, Wormser GP. 2013. CTX-M -lactamaseproducing Klebsiella pneumoniae in suburban New York city, New York, USA. Emerg Infect Dis 19:1803–1810. http://dx.doi.org/10.3201/eid1911 .121470. Richter SN, Frasson I, Franchin E, Bergo C, Lavezzo E, Barzon L, Cavallaro A, Palù G. 2012. KPC-mediated resistance in Klebsiella pneumoniae in two hospitals in Padua, Italy, June 2009 to December 2011: massive spreading of a KPC-3-encoding plasmid and involvement of nonintensive care units. Gut Pathog 4:7. Giakkoupi P, Papagiannitsis CC, Miriagou V, Pappa O, Polemis M, Tryfinopoulou K, Tzouvelekis LS, Vatopoulos AC. 2011. An update of the evolving epidemic of blaKPC-2-carrying Klebsiella pneumoniae in Greece (2009 to 2010). J Antimicrob Chemother 66:1510 –1513. http://dx .doi.org/10.1093/jac/dkr166. Lascols C, Peirano G, Hackel M, Laupland KB, Pitout JD. 2013. Surveillance and molecular epidemiology of Klebsiella pneumoniae isolates that produce carbapenemases: first report of OXA-48-like enzymes in North America. Antimicrob Agents Chemother 57:130 –136. http://dx.doi .org/10.1128/AAC.01686-12.
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blaNDM-5-Bearing IncFII-Type Plasmids of Klebsiella pneumoniae Sequence Type 147 Transmitted by Cross-Border Transfer of a Patient Juyoun Shin,a Jin Yang Baek,b Sun Young Cho,d Hee Jae Huh,c Nam Yong Lee,c Jae-Hoon Song,b,d Doo Ryeon Chung,b,d Kwan Soo Koa,b Department of Molecular Cell Biology, Sungkyunkwan University School of Medicine, Suwon, South Koreaa; Asia Pacific Foundation for Infectious Diseases (APFID), Seoul, South Koreab; Department of Laboratory Medicine, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, South Koreac; Division of Infectious Diseases, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, South Koread
The two plasmids extracted from Klebsiella pneumoniae sequence type 147 (ST147) isolates were analyzed. The first isolate was obtained from a patient transferred from United Arab Emirates to South Korea. The second isolate was obtained from a Korean patient and was suspected to be transmitted from the first patient. Sequences of two plasmids were almost the same, and genetic structures, including blaNDM-5, of these plasmids were similar to plasmids of NDM-1-producing Escherichia coli ST131 isolates found in Europe.
T
hus, far, 13 variants of NDM have been identified (1). Of these, NDM-5 has been repeatedly isolated, mainly from Escherichia coli found in European, Asian, and African countries (2–7). Recently, two NDM-5-producing Klebsiella pneumoniae isolates (CC1409-1 and CC1410-1) were identified in South Korea (8). The first isolate was collected from a patient who was transferred from Abu Dhabi, United Arab Emirates (UAE), to Seoul, South Korea, and the second isolate was collected from a Korean patient after 4 months. Because both the isolates belonged to the same clone and shared many antimicrobial resistance genes, the second isolate was thought to have been transmitted by cross-border transfer from the patient from UAE (8). In the present study, we determined the complete sequences of the two blaNDM-5-bearing plasmids (pCC1409-1 and pCC1410-1) isolated from the K. pneumoniae isolates. We confirmed that the two plasmids had similar sequences, with minor differences, and were very similar to blaNDM-1-bearing IncFII-type plasmids of E. coli isolates found in Europe. These plasmids were conjugated using E. coli J53, a sodium azide-resistant and plasmid-free strain, as a recipient, as in the previous study (9). The MiSeq desktop sequencer system (Illumina, Inc., San Diego, CA) was used to determine the complete sequence of the plasmids pCC1409-1 and pCC1410-1. A total of 3,768,155 and 3,825,823 sequence reads were generated, yielding an average sequencing depth of 11,845⫻ and 13,312⫻ for the plasmids pCC1409-1 and pCC1410-1, respectively. PCR combinations were used to close gaps between the contigs and to confirm the position of the contigs (see the supplemental material). Open reading frames were predicted, annotated using the Glimmer 3.0 system (http://ccb.jhu.edu/software/glimmer /index.shtml), and confirmed using DNAMAN 5.2.10 software (Lynnon BioSoft, Lynnon Corporation). Each predicted protein was compared against an all-protein database by using BLASTP (http://blast.ncbi.nlm.nih.gov/blast.cgi). Gene sequences were compared and aligned with those of genes in pGUE-NDM and pMC-NDM by using cross_match 1.080812 software and ClustalW (http://www.ebi.ac.uk/Tools/msa/clustalw2). Whole-plasmid sequencing indicated that plasmid pCC1409-1 isolated from the K. pneumoniae CC1409-1 isolate collected from the patient transferred from UAE was 89,745 bp in size and had 114 protein coding sequences (CDS). The other plasmid, pCC1410-1, of the K. pneumoniae CC1410-1 isolate col-
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lected from the Korean patient was 95,005 bp in size and was predicted to harbor 120 CDS. Both of the plasmids belonged to the incompatibility group IncFII, with their G⫹C contents being 53.04% and 52.96%, respectively. Comparison of their genetic structures indicated that these plasmids carried almost the same genes, except that pCC1410-1 had an ISCR3 insertion sequence containing erm genes that encoded rRNA adenine N-6-methyltransferase (Fig. 1). The strain isolated from the first patient may have been transmitted to the second patient through unknown transmitters, hospital equipment, hospital staffs, or other patients. During the transmission, a small downstream region of blaNDM-5 may have been incorporated into the second plasmid, pCC1410-1. Comparison of databases indicated that pCC1409-1 and pCC1410-1 shared 99% sequence similarity with both pGUENDM (GenBank accession no. JQ364967.1) and pMC-NDM (GenBank accession no. HG003695.1) carrying NDM-1 genes, which were isolated from carbapenem-resistant E. coli ST131 isolates found in France and Poland, respectively (Fig. 1) (10, 11). In particular, backbone structures, including replication, partition, and conjugative transfer loci, of pCC1409-1 and pCC1410-1 showed very high nucleotide similarity with those of pGUE-NDM and pMC-NDM. All of the 24 tra and 8 trb genes, which contributed to plasmid transferability of pCC1409-1 and pCC1410-1, showed 97% and 95% sequence similarity, respectively, to those in pGUE-NDM and pMC-NDM. These transfer loci of pCC1409-1 and pCC1410-1 were more similar to those of blaCTX-M-15-bearing
Received 10 November 2015 Returned for modification 6 December 2015 Accepted 21 December 2015 Accepted manuscript posted online 11 January 2016 Citation Shin J, Baek JY, Cho SY, Huh HJ, Lee NY, Song J-H, Chung DR, Ko KS. 2016. blaNDM-5-bearing IncFII-type plasmids of Klebsiella pneumoniae sequence type 147 transmitted by cross-border transfer of a patient. Antimicrob Agents Chemother 60:1932–1934. doi:10.1128/AAC.02722-15. Address correspondence to Doo Ryeon Chung, [email protected], or Kwan Soo Ko, [email protected]. Supplemental material for this article may be found at http://dx.doi.org/10.1128 /AAC.02722-15. Copyright © 2016, American Society for Microbiology. All Rights Reserved.
Antimicrobial Agents and Chemotherapy
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blaNDM-5-Bearing K. pneumoniae Plasmids
FIG 1 Comparison of the major structural features of pCC1409-1 and pCC1410-1 with those of IncFII-type plasmids pGUE-NDM (GenBank accession no. JQ364967.1) and pMC-NDM (GenBank accession no. HG003695.1). White boxes indicate plasmid scaffold regions that are common among the plasmids. The tra loci are indicated within the white boxes by capital letters. Resistance genes are indicated by gray boxes, and transposon-related genes, integrase, and insertion sequences are indicated by diagonally lined boxes. Replicase genes are indicated by dots, and addiction system genes are indicated in black. Light gray shading denotes regions of homology, and arrows indicate blaNDM-type genes.
pC15-1a (GenBank accession no. AY458016.1) and pHK23a (GenBank accession no. JQ432559.1) isolated from E. coli isolates found in Canada and Hong Kong, respectively (12, 13). In addition, both pCC1409-1 and pCC1410-1 contained identical IncFII replicons repA2-repA6-repA1-repA4, a 2,166-bp fragment (G⫹C content, 51.89%; dotted boxes in Fig. 1), that showed high nucleotide similarity to IncFII replicons of pMCNDM and pGUE-NDM. pCC1409-1 and pCC1410-1 contained 11 and 13 antimicrobial resistance genes, respectively, including a blaNDM-type-bleMBL-trpF-tat module and the -lactamase gene blaTEM-1 (Fig. 1). In addition, genes conferring resistance against macrolides [mph(A), mrx, and mphR(A)], sulfonamides (sul1), quaternary ammonium compounds (qacE⌬1), aminoglycosides (aadA4 and rmtB), and trimethoprim (dfrA17) were identified in the antimicrobial resistance region of pCC1409-1 and pCC1410-1. The upstream and downstream structures of blaNDM-5 were almost identical to those of blaNDM-1 in pMC-NDM (Fig. 1). NDM-5 differs from NDM-1 by two amino acid substitutions at positions 88 (Val¡Leu) and 154 (Met¡Leu) (2). The blaNDM-5bearing plasmids examined in the present study had the same origin as pMC-NDM. However, the proximal part of the resistance region [lqsb]from insAB to mphR(A)] in pCC1409-1 and pCC1410-1 was different from that in pGUE-NDM and pMCNDM (Fig. 1). The adjacent structure of blaNDM-5 has been identified in other NDM-5-producing E. coli isolates. Although complete sequences of plasmids of these E. coli isolates have not been determined, the proximal structure of their resistance region is suggested to be different from that of plasmids examined in the present study (3, 5). Although blaNDM-5-bearing pCC1409-1 and pCC1040-1 had the same origin as blaNDM-1-bearing pMC-NDM, the proximal part of their resistance region might have originated from other plasmids. Further, blaNDM-5-containing region of pCC1409-1 and pCC1410-1 differed from that of a plasmid isolated from NDM1-producing K. pneumoniae isolates found in South Korea (14).
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Both pCC1409-1 and pCC1410-1 were isolated from K. pneumoniae ST147 isolates. K. pneumoniae ST147 isolates that are resistant to diverse antibiotics have disseminated worldwide (15– 19); however, ST147 isolates, to our knowledge, have not been found in K. pneumoniae from South Korea. Therefore, we suggest that pCC1409-1 and pCC1410-1 were not derived from K. pneumoniae isolates found in South Korea. In this study, we determined the complete sequences of the two IncFII-type plasmids isolated from NDM-5-producing K. pneumoniae isolates and observed that the two plasmids had very similar sequences. To our knowledge, this is the first study to report the complete sequences of blaNDM-5-bearing plasmids. Nucleotide sequence accession numbers. The annotated sequences of pCC1409-1 and pCC1410-1 have been submitted to GenBank nucleotide sequence database under accession numbers KT725789 and KT725788, respectively. ACKNOWLEDGMENTS This research was supported by the Basic Science Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT, and Future Planning (grant NRF2013R1A2A2A0104103). J. Shin was partly supported by the Basic Science Research Program through the NRF funded by the Ministry of Education (grant 2013R1A12062884).
REFERENCES 1. Shrestha B, Tada T, Miyoshi-Akiyama T, Shimada K, Ohara H, Kirikae T, Pokhrel BM. 2015. Identification of a novel variant, NDM-13, from a multidrug-resistant Escherichia coli from Nepal. Antimicrob Agents Chemother 59:5847–5850. http://dx.doi.org/10.1128/AAC.00332-15. 2. Hornsey M, Phee L, Wareham DW. 2011. A novel variant, NDM-5, of the New Delhi metallo--lactamase in a multidrug-resistant Escherichia coli ST648 isolate recovered from a patient in the United Kingdom. Antimicrob Agents Chemother 55:5952–5954. http://dx.doi.org/10.1128/AAC .05108-11. 3. Yang P, Xie Y, Feng P, Zong Z. 2014. blaNDM-5 carried by an IncX3 plasmid in Escherichia coli sequence type 167. Antimicrob Agents Chemother 58:7548 –7552. http://dx.doi.org/10.1128/AAC.03911-14.
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Antimicrobial Agents and Chemotherapy
March 2016 Volume 60 Number 3
