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Citrobacter spp. simultaneously harboring blaCTX-M, blaTEM, blaSHV, blaampC, and insertion sequences IS26 and orf513: an evolutionary phenomenon of recent concern for antibiotic resistance.

ABSTRACT: A collection of 40 confirmed isolates of Citrobacter spp., obtained from the culture of 5,732 clinical samples during 2006 to 2008, was studied for the presence of blaCTX-M, blaTEM, blaSHV, and blaampC by monoplex and multiplex PCRs. These isolates were also looked at for the presence of insertion elements IS26 and orf513, which play an important role in the resistance gene pool. blaCTX-M, blaTEM, blaSHV, and blaampC were noticed in 67.5%, 40%, 25%, and 40% of isolates, respectively. Sequencing for the specific CTX-M type revealed the presence of CTX-M-15-type extended-spectrum beta-lactamases (ESBL). sul1-type integrons were detected in 32.5% (13/40) of isolates, and a single plasmid of ca. approximately 22 kb was also noticed in all the isolates harboring bla genes. A total of 48.2% (13/27) of isolates harboring blaCTX-M alleles were found to carry IS26 elements and 53.4% (7/13) of isolates harboring the sul1-type integrons were found to carry orf513. Alarmingly, a few isolates simultaneously carried IS26 and orf513, reflecting the occurrence of complex evolution. All the Citrobacter isolates were noticed to produce an amplicon of 400 bp in an IS26 PCR, similar to that of the predominant UK CTX-M-15-producing Escherichia coli clone (clone A) and suggesting a probable genetic relatedness between the two, which could not be proved with British and Indian E. coli isolates in our earlier studies. Here, a first report is being presented describing the occurrence of blaCTX-M-15 in Indian Citrobacter spp. This is also the first report describing the simultaneous occurrence of blaTEM, blaSHV, and blaampC along with blaCTX-M in class 1 integrons. The recent findings indicate that the genetic environment of blaCTX-M-15 has changed through the insertion of IS26 and orf513, although the consequences of these events remain uncertain. It perhaps also suggests a "turnover" of these mobile elements in the population over time.

SUBMITTER: Shahid M

PROVIDER: S-EPMC2863875 | biostudies-literature | 2010 May

REPOSITORIES: biostudies-literature

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Citrobacter spp. simultaneously harboring blaCTX-M, blaTEM, blaSHV, blaampC, and insertion sequences IS26 and orf513: an evolutionary phenomenon of recent concern for antibiotic resistance.

Shahid M M

Journal of clinical microbiology 20100310 5

A collection of 40 confirmed isolates of Citrobacter spp., obtained from the culture of 5,732 clinical samples during 2006 to 2008, was studied for the presence of blaCTX-M, blaTEM, blaSHV, and blaampC by monoplex and multiplex PCRs. These isolates were also looked at for the presence of insertion elements IS26 and orf513, which play an important role in the resistance gene pool. blaCTX-M, blaTEM, blaSHV, and blaampC were noticed in 67.5%, 40%, 25%, and 40% of isolates, respectively. Sequencing ...[more]

PMID: 20220171

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Project description:Multidrug-resistant (MDR) patterns due to extended-spectrum β-lactamase (ESBL) production in pathogenic bacteria are now becoming prevalent in hospitals worldwide, posing a public health challenge. The aim of the present study was to determine the antibiotic susceptibility patterns and distribution of the blaTEM, blaCTX-M, blaSHV and blaOXA ESBL resistance genes in MDREnterobacteriaceae and Acinetobacter baumannii (A. baumannii). A cross-sectional study was conducted between September 2017 and August 2018 in the King Abdullah Hospital (Bisha, Saudi Arabia). Bacterial isolates were collected from the clinical samples of patients; these were identified and screened for ESBL production and their antibiotic susceptibility was examined using standard microbiology methods. Multiplex-PCR runs were performed to identify genes encoding ESBL producers. DNA sequencing analysis was used to identify the specific gene variants. Of the 274 isolates, 173 (63.1%) exhibited MDR patterns to different antibiotics. A. baumannii revealed the highest resistance rates for cefuroxime (100%), gentamicin (88%) and amikacin (86%). Klebsiella pneumoniae (K. pneumoniae) isolates had the highest resistance rates for cefuroxime (98%), aztreonam and trimethoprim/sulfamethoxazole (87% for each). Escherichia coli (E. coli) exhibited high resistance rates for trimethoprim/sulfamethoxazole (92%) and cefuroxime (87%). Of the 173 MDR isolates, 78 (45.1%) exhibited ESBL production. Of these, 88.9% (72/78) carried ESBL genes. The most prevalent gene-encoding isolates were blaTEM (84.7%), followed by blaCTX-M (33.3%), blaSHV (2.7%) and blaOXA-1 (1.4%). A single blaTEM gene was predominantly produced by K. pneumoniae (60.7%), A. baumannii (78.9%) and Proteus mirabilis (80%), whereas blaCTX-M was harbored by E. coli (33.3%). The co-existence of two different genes in a single bacterium was revealed in 22.2% of isolates, commonly between blaTEM and blaCTX-M (19.4%). Sequencing analysis revealed that blaCTX-M-15 and blaTEM-1 were predominant variants of the blaCTX-M and blaTEM genes, respectively. The present study revealed a diversity of ESBL genes in Gram-negative bacterial isolates, with blaTEM being the most prevalent type. The emergence of various ESBL genes with several co-existing genotypes is alarming, rendering extensive surveillance studies necessary to understand the transmission and epidemiology of such resistant gene-carrying isolates.

Project description:UnlabelledCarbapenemase-producing Enterobacteriaceae (CPE), which are resistant to most or all known antibiotics, constitute a global threat to public health. Transposable elements are often associated with antibiotic resistance determinants, suggesting a role in the emergence of resistance. One insertion sequence, IS26, is frequently associated with resistance determinants, but its role remains unclear. We have analyzed the genomic contexts of 70 IS26 copies in several clinical and surveillance CPE isolates from the National Institutes of Health Clinical Center. We used target site duplications and their patterns as guides and found that a large fraction of plasmid reorganizations result from IS26 replicative transpositions, including replicon fusions, DNA inversions, and deletions. Replicative transposition could also be inferred for transposon Tn4401, which harbors the carbapenemase blaKPC gene. Thus, replicative transposition is important in the ongoing reorganization of plasmids carrying multidrug-resistant determinants, an observation that carries substantial clinical and epidemiological implications for understanding how such extreme drug resistance phenotypes evolve.ImportanceAlthough IS26 is frequently reported to reside in resistance plasmids of clinical isolates, the characteristic hallmark of transposition, target site duplication (TSD), is generally not observed, raising questions about the mode of transposition for IS26. The previous observation of cointegrate formation during transposition implies that IS26 transposes via a replicative mechanism. The other possible outcome of replicative transposition is DNA inversion or deletion, when transposition occurs intramolecularly, and this would also generate a specific TSD pattern that might also serve as supporting evidence for the transposition mechanism. The numerous examples we present here demonstrate that replicative transposition, used by many mobile elements (including IS26 and Tn4401), is prevalent in the plasmids of clinical isolates and results in significant plasmid reorganization. This study also provides a method to trace the evolution of resistance plasmids based on TSD patterns.

Dataset Information

Citrobacter spp. simultaneously harboring blaCTX-M, blaTEM, blaSHV, blaampC, and insertion sequences IS26 and orf513: an evolutionary phenomenon of recent concern for antibiotic resistance.

Publications

Citrobacter spp. simultaneously harboring blaCTX-M, blaTEM, blaSHV, blaampC, and insertion sequences IS26 and orf513: an evolutionary phenomenon of recent concern for antibiotic resistance.

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