Molecular Analysis of the Antibiotic Resistant NDM-1 Gene in Clinical Isolates of Enterobacteriaceae

Ejaz, Hasan; Alzahrani, Badr; Hamad, Mutaz F S; Abosalif, Khalid Omer Abdalla; Junaid, Kashaf; Abdalla, Abualgasim Elgaili; Elamir, Mohammed Yagoub Mohammed; Aljaber, Nawaf J; Hamam, Sanaa S.; Younas, Sonia

doi:10.7754/clin.lab.2019.190727

Cited by 18 publications

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“…Polymyxin B and colistin have saved patients' lives and are considered a vital regimen for treating XDR bacterial infections [12]. Other studies have reported the use of colistin, aminoglycosides, co-trimoxazole, piperacillin-tazobactam, cefoperazone-sulbactam, and tigecycline to treat multidrug-resistant bacteria [18]. Unfortunately, Col-R isolates have emerged worldwide because of the injudicious use of colistin, particularly in veterinary medicine [5].…”

Section: Discussionmentioning

confidence: 99%

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Emergence of plasmid-mediated mcr genes from Gram-negative bacteria at the human-animal interface

Javed

Zafar²,

Ghafoor

et al. 2020

Gut Pathog

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Background The global emergence of plasmid-mediated colistin resistance (Col-R) conferred by mcr genes in gram-negative rods (GNRs) has jeopardized the last treatment option for multidrug-resistant bacterial infections in humans. This study aimed to assess the emergence of mcr gene-mediated Col-R in GNRs isolated from humans and animals in Pakistan. Methods Animal and clinical specimens collected from various sources were prospectively analysed using standard microbiological procedures. Pathogens were identified using the API 20E and API 20NE systems (bioMerieux). Minimum inhibitory concentration (MIC) against colistin was determined using the MIC detection methods, and multiplex polymerase chain reaction (PCR) was used to amplify the mcr-1 to mcr-5 genes. Results We isolated 126 (88.1%) animal and 17 (11.9%) human Col-R phenotypes, among which there was a significant association (P < 0.01) of Escherichia coli and Proteus mirabilis with animals and of Acinetobacter baumannii with humans. Animal strains exhibited statistically significant (P < 0.05) resistance to co-trimoxazole, chloramphenicol, and moxifloxacin, and the human pathogens exhibited statistically significant (P < 0.05) antibiotic resistance to cephalosporins, carbapenems, and piperacillin-tazobactam. For Col-R strains, MIC50 values were > 6 µg/mL and > 12 µg/mL for human and animal isolates, respectively. mcr genes were detected in 110 (76.9%) bacterial strains, of which 108 (98.2%) were mcr-1 and 2 (1.8%) were mcr-2. Conclusions The detection of a considerable number of mcr-1 and mcr-2 genes in animals is worrisome, as they are now being detected in clinical pathogens. The acquisition of mcr genes by colistin-susceptible bacteria could leave us in a post-antibiotic era.

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Section: Discussionmentioning

confidence: 99%

“…The bacterial DNA from the freshly cultured GNR strains was thermally extracted by emulsifying 2-3 colonies in 200 µL of Tris EDTA (TE) buffer and boiling for 10 min [18]. Previously described mcr1 to mcr5 primers were used in the multiplex polymerase chain reaction (PCR) [19].…”

Section: Molecular Detection Of Mcr Genes and Data Analysismentioning

confidence: 99%

Emergence of plasmid-mediated mcr genes from Gram-negative bacteria at the human-animal interface

Javed

Zafar²,

Ghafoor

et al. 2020

Gut Pathog

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“…The DNA from each specimen was extracted by the boiling method from the lysed bacterial strains, and concentration was measured using a NanoDrop spectrophotometer. The supernatant was stored at -20˚C and used for the amplification of the genes [17].…”

Section: Dna Extraction and Molecular Analysismentioning

confidence: 99%

Molecular analysis of blaSHV, blaTEM, and blaCTX-M in extended-spectrum β-lactamase producing Enterobacteriaceae recovered from fecal specimens of animals

et al. 2021

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Colonization of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae as animal gut microbiota is a substantial global threat. This study aimed to determine the molecular characterization of blaSHV, blaTEM, and blaCTX-M variants in animals, as well as to evaluate the antimicrobial resistance conferred by these genes. We prospectively analyzed 1273 fecal specimens of farm and domestic animals for the isolation of enterobacteria that had the ESBL phenotype by using biochemical methods. The extracted genes were amplified by polymerase chain reaction and sequenced for the characterization of blaSHV, blaTEM, and blaCTX-M variants. The drug-resistance spectrum and hierarchical clusters were analyzed against 19 antibacterial agents. Out of 245 (19.2%) ESBL enterobacteria, 180 (75.5%) Escherichia coli and 34 (13.9%) Klebsiella pneumoniae were prevalent species. A total of 73.9% blaCTX-M, 26.1% blaTEM, and 14.2% blaSHV were found among the enterobacteria; however, their association with farm or domestic animals was not statistically significant. The distribution of bla gene variants showed the highest number of blaCTX-M-1 (133; 54.3%), followed by blaCTX-M-15 (28; 11.4%), blaTEM-52 (40; 16.3%), and blaSHV-12 (22; 9%). In addition, 84.5% of the enterobacteria had the integrons intI1. We observed ±100% enterobacteria resistant to cephalosporin, 7 (2.9%) to colistin (minimum inhibitory concentration breakpoint ≥4 μg/mL), 9 (3.7%) to piperacillin-tazobactam, 11 (4.5%) to imipenem, 14 (5.7%) to meropenem, and 18 (7.3%) to cefoperazone-sulbactam, without statistically significant association. Animal gut microbiota contain a considerable number of blaCTX-M, blaTEM, blaSHV, and integrons, which are a potential source of acquired extensive drug resistance in human strains and leaves fewer therapeutic substitutes.

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“…The plasmids containing mcr-1 have been identified in MDR Enterobacteriaceae isolates, including species that produce carbapenemases, such as K. pneumoniae carbapenemase (KPC), Verona integron-encoded metallo-β-lactamase (MBL)-producing (VIM) species, or New Delhi MBL (NDM)-producing species [ 12 ]. Since the discovery of NDM-1 in India, over 24 NDM variants have been identified [ 13 ]. The co-existence of mcr and NDM has been reported in specimens isolated from food-producing animals and clinical samples, increasing the public health burden associated with antimicrobial resistance [ 14 ].…”

Section: Introductionmentioning

confidence: 99%

Molecular Epidemiology of Extensively Drug-Resistant mcr Encoded Colistin-Resistant Bacterial Strains Co-Expressing Multifarious β-Lactamases

et al. 2021

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Plasmid-mediated colistin resistance (Col-R) conferred by mcr genes endangers the last therapeutic option for multifarious β-lactamase-producing bacteria. The current study aimed to explore the mcr gene molecular epidemiology in extensively drug-resistant (XDR) bacteria. Col-R gram-negative bacterial strains were screened using a minimum inhibitory concentration (MIC) breakpoint ≥4 µg/mL. Resistant isolates were examined for mcr variants, extended-spectrum β-lactamase, AmpC, and carbapenemase genes using polymerase chain reaction (PCR). The MIC breakpoints for mcr-positive strains were determined using broth microdilution and E-test strips. Overall, 19/718 (2.6%) gram-negative rods (GNRs) harboring mcr were identified, particularly in pus (p = 0.01) and tracheal secretions (p = 0.03). Molecular epidemiology data confirmed 18/19 (95%) mcr-1 and 1/19 (5%) mcr-2 genes. Integron detection revealed 15/17 (88%) Int-1 and 2/17 (12%) Int-2. Common co-expressing drug-resistant β-lactamase genes included 8/16 (50%) blaCTM-1, 3/16 (19%) blaCTM-15, 3/3 (100%) blaCMY-2, 2/8 (25%) blaNDM-1, and 2/8 (25%) blaNDM-5. The MIC50 and MIC90 values (µg/mL) were as follows: Escherichia coli, 12 and 24; Klebsiella pneumoniae, 12 and 32; Acinetobacter baumannii, 8 and 12; and Pseudomonas aeruginosa, 32 and 64, respectively. Treatment of XDR strains has become challenging owing to the co-expression of mcr-1, mcr-2, multifarious β-lactamase genes, and integrons.

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Molecular Analysis of the Antibiotic Resistant NDM-1 Gene in Clinical Isolates of Enterobacteriaceae

Cited by 18 publications

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Emergence of plasmid-mediated mcr genes from Gram-negative bacteria at the human-animal interface

Emergence of plasmid-mediated mcr genes from Gram-negative bacteria at the human-animal interface

Molecular analysis of blaSHV, blaTEM, and blaCTX-M in extended-spectrum β-lactamase producing Enterobacteriaceae recovered from fecal specimens of animals

Molecular Epidemiology of Extensively Drug-Resistant mcr Encoded Colistin-Resistant Bacterial Strains Co-Expressing Multifarious β-Lactamases

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