Dataset Information

Separation and confirmation of nine Enterobacteriaceae strains that carry the blaNDM-1 gene.

ABSTRACT: The aim of the present study was to confirm the existence of carbapenem-resistant Enterobacteriaceae carrying the blaNDM-1 gene in clinics in Hainan province, China. Collected clinical bacterial isolates that were Enterobacteriaceae strains suspected of producing carbapenemase were used as experimental strains. Drug resistance to imipenem, meropenem and other antibacterial agents was tested. Imipenem/imipenem inhibitor (IP/IPI) E-testing was conducted to identify the bacterial strains that produced metallo-?-lactamases. The blaNDM-1 drug resistance gene was amplified by polymerase chain reaction (PCR), and agarose gel electrophoresis (AGE) and sequencing were conducted to identify the products. The species of the strains carrying the blaNDM-1 gene were determined using a biochemical identification system. Through the IP/IPI E-test, 21 of the 30 collected Enterobacteriaceae strains were found to be positive, indicating that 70% of the strains produced metallo-?-lactamases. Following blaNDM-1 gene PCR amplification, AGE and sequencing tests confirmed that nine of the strains carried the blaNDM-1 drug resistance gene. The biochemical identification system indicated that four of the strains were Klebsiella pneumoniae, two were Escherichia coli, two were Enterobacter cloacae and one was Enterobacter aerogenes. Drug susceptibility testing in vitro demonstrated that the strains were 100% resistant to a broad spectrum antibiotic plus lactamase inhibitor, cephalosporins and carbapenems. However, they had high sensitivity rates to polymyxin B and tigecycline of 100 and 88.9%, respectively. The sensitivity rate to amikacin was also high at 77.8%, whereas sensitivity to ciprofloxacin and gentamicin was moderate at rates of 44.4 and 33.3% respectively. This clinical study of Enterobacteriaceae strains that carry the blaNDM-1 gene in Hainan shows a bacterial tolerance that is different from that in previous studies, which requires further in-depth study.

SUBMITTER: Li TJ

PROVIDER: S-EPMC4353756 | biostudies-literature | 2015 Apr

REPOSITORIES: biostudies-literature

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Separation and confirmation of nine Enterobacteriaceae strains that carry the blaNDM-1 gene.

Li Tian-Jiao TJ Li Chen-Xue CX Cheng Shu-Ping SP Wang Xu-Ming XM Fu Sheng-Miao SM Li Xiao-Juan XJ Huang Tao T Fu Hui-Qun HQ Lin Song S Lu Y E YE

Experimental and therapeutic medicine 20150203 4

The aim of the present study was to confirm the existence of carbapenem-resistant Enterobacteriaceae carrying the blaNDM-1 gene in clinics in Hainan province, China. Collected clinical bacterial isolates that were Enterobacteriaceae strains suspected of producing carbapenemase were used as experimental strains. Drug resistance to imipenem, meropenem and other antibacterial agents was tested. Imipenem/imipenem inhibitor (IP/IPI) E-testing was conducted to identify the bacterial ...[more]

PMID: 25780416

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Project description:Objective:Regional dissemination is the major cause of the widespread prevalence of a plasmid-encoding NDM-1 enzyme. We investigated the drug resistance, joint efficiency, and gene environment of a Klebsiella pneumoniae strain carrying bla NDM-1 gene. Materials and Methods:Carbapenem-non-susceptible strains were analyzed using the VITEK 2 Compact. Strains carrying bla NDM-1 were identified using polymerase chain reaction and sequencing. Antimicrobial susceptibility testing and plasmid conjugation experiments were then conducted. Strains carrying bla NDM-1 were subjected to Southern blot analysis. After the gene mapping of bla NDM-1, library construction, and sequencing, plasmids were subsequently spliced and genotyped using the software Glimmer 3.0, and then analyzed using Mauve software. Results:Among 1735 carbapenem-non-susceptible strains, 54 strains of bla NDM-1-positive bacteria were identified, which consisted of 44 strains of K. pneumoniae, 8 strains of Acinetobacter baumannii and 2 strains of Escherichia coli. Strains carrying bla NDM-1 had a resistance rate of more than 50% in most antibiotics. Plasmid conjugation between strains carrying bla NDM-1 and E. coli strain J53 had a success rate of 50%. Southern blot analysis indicated that each strain had multiple plasmids containing bla NDM-1. Among the five plasmids containing bla NDM-1 in K. pneumoniae for sequencing, two plasmids with complete sequences were obtained. The findings were as follows: (i) The p11106 and p12 plasmids were highly similar to pNDM-BTR; (ii) the p11106 and p12 plasmids showed differences in the 20-30 kb region (orf00032-orf00043) from the other six plasmids; and (iii) bla NDM-1 was located at orf00037, while ble was found at orf00038. Two tnpA genes were located in the upstream region, and orf00052 (tnpA) in the 36 kb region was in the downstream sequence. Conclusion:bla NDM-1-containing bacteria exhibit multidrug resistance, which rapidly spreads and is transferred through efficient plasmid conjugation; the multidrug resistance of these bacteria may be determined by analyzing their drug-resistant plasmids. The presence of ble and tnpA genes suggests a possible hypothesis that bla NDM-1 originates from A. baumannii, which is retained in K. pneumoniae over a long period by transposition of mobile elements.

Project description:BackgroundThe bla NDM-1 (New Delhi Metallo-β-lactamase-1) gene has disseminated around the globe. NDM-1 producers are found to co-harbour resistance genes against many antimicrobials, including fluoroquinolones. The spread of large plasmids, carrying both bla NDM and plasmid-mediated fluoroquinolone resistance (PMQR) markers, is one of the main reasons for the failure of these essential antimicrobials.MethodsEnterobacteriaceae (n = 73) isolated from the blood of septicaemic neonates, admitted at a neonatal intensive care unit (NICU) in Kolkata, India, were identified followed by PFGE, antibiotic susceptibility testing and determination of MIC values for meropenem and ciprofloxacin. Metallo-β-lactamases and PMQRs were identified by PCR. NDM-positive isolates were studied for mutations in GyrA & ParC and for co-transmission of bla NDM and PMQR genes (aac(6')-Ib-cr, qnrB, qnrS) through conjugation or transformation. Plasmid types, integrons, plasmid addiction systems, and genetic environment of the bla NDM gene in NDM-positive isolates and their transconjugants/ transformants were studied.ResultsIsolated Enterobacteriaceae comprised of Klebsiella pneumoniae (n = 55), Escherichia coli (n = 16), Enterobacter cloacae (n = 1) and Enterobacter aerogenes (n = 1). The rates of ciprofloxacin (90%) and meropenem (49%) non-susceptibility were high. NDM was the only metallo-β-lactamase found in this study. NDM-1 was the predominant metallo-β-lactamase but NDM-5, NDM-7, and NDM-15 were also found. There was no significant difference in ciprofloxacin non-susceptibility (97% vs 85%) and the prevalence of PMQRs (85% vs 77%) between NDM-positive and NDM-negative isolates. Among the PMQRs, aac(6')-Ib-cr was predominant followed by qnrB1 and qnrS1. Twenty-nine isolates (40%) co-harboured PMQRs and bla NDM, of which 12 co-transferred PMQRs along with bla NDM in large plasmids of IncFIIK, IncA/C, and IncN types. Eighty-two percent of NDM-positive isolates possessed GyrA and/or ParC mutations. Plasmids carrying only bla NDM were of IncHIB-M type predominantly. Most of the isolates had ISAba125 in the upstream region of the bla NDM gene.ConclusionWe hypothesize that the spread of PMQRs was independent of the spread of NDM-1 as their co-transfer was confirmed only in a few isolates. However, the co-occurrence of these genes poses a great threat to the treatment of neonates.

Project description:Aim of this study was to genetically characterize two carbapenemase-producing Escherichia coli strains obtained from a pediatric patient affected by diarrhea, expressing OXA-181 and/or NDM-5 type enzymes. The above microorganisms were collected in the same Desenzano hospital (Northern Italy) where the bla NDM- 5 gene was detected for the first time in Italy 3 years ago. One strain (5P), belonged to sequence type ST405/ST477 (according to Pasture/Oxford schemes) and serotype O102:H6. It was characterized by a 130562 bp multi-replicon plasmid IncFII/IncFIA/IncFIB (pVSI_NDM-5) enclosing two main antibiotic resistance islands: (i) ARI-I, 10030 bp in size, carried genes coding for β-lactam- (bla OXA- 1, bla CTX-M- 15), fluoroquinolone/aminoglycoside- (aac(6')-lb-cr) and phenicol- resistance (catB3), (ii) ARI-II, 15326 bp in size, carried genes coding for sulfonamide- (sul1), β-lactam- (bla NDM- 5, bla TEM- 1 B), phenicol- (catB3), trimethoprim- (dfrA17), antiseptic- (qacEΔ1), and aminoglycoside- (aadA5, rmtB) resistance. The other isolate (5M), belonged to sequence type ST2659/ST759 and serotype O50/02:H18, and carried four plasmids: a 153866 bp multi-replicon IncFII/IncFIA/IncFIB (pISV_IncFII_NDM-5), an 89866 bp IncI1 plasmid, a 51480 bp IncX3 plasmid (pISV_IncX3_OXA181), and a 41143 bp IncI plasmid (pISV_IncI_CMY-42). pISV_IncFII_NDM-5 carried two main antibiotic resistance islands: (i) ARI-III, 12220 bp in size, carried genes coding for β-lactam- (bla OXA- 1), fluoroquinolone/aminoglycoside- (aac(6')-lb-cr), tetracycline- (tet(B)) and phenicol- resistance (catB3, catA1), and ii) ARI-IV, 26527 bp in size, carried determinants coding for macrolide- (erm(B), mph(A)), sulfonamide- (sul1), beta-lactam- (bla NDM- 5, bla TEM- 1 B), trimethoprim- (dfrA14, dfrA12), antiseptic- (qacEΔ1), and aminoglycoside- resistance (aadA5). pISV_IncI_CMY-42 harbored the bla CMY- 42 gene coding for beta-lactam resistance, pISV_IncX3_OXA181 harbored genes encoding fluoroquinolone- (qnrS1) and beta-lactams- resistance (bla OXA- 181). In conclusion, the detection of two different NDM-5 E. coli strains from a pediatric patient with a history of travel to the Far East countries strongly highlight an increasing trend and risk of importation from such areas.

Dataset Information

Separation and confirmation of nine Enterobacteriaceae strains that carry the blaNDM-1 gene.

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Separation and confirmation of nine Enterobacteriaceae strains that carry the <i>bla</i><sub>NDM-1</sub> gene.

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