- Rapid determination of carbapenem resistance by low-cost colorimetric methods: Propidium Iodide and alamar blue staining
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Jiyoon Choi , Jiwon Baek , Daehyuk Kweon , Kwan Soo Ko , Hyunjin Yoon
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J. Microbiol. 2020;58(5):415-421. Published online March 28, 2020
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DOI: https://doi.org/10.1007/s12275-020-9549-x
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Abstract
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Carbapenems are a class of β-lactam antibiotics with a broad
antimicrobial activity spectrum. Owing to their sturdy structures
resistant to most β-lactamases, they have been regarded
as one of the last-resort antibiotics for combating multidrugresistant
bacterial infections. However, the emergence of carbapenem
resistance increases predominantly in nosocomial
pathogens. To prevent spread of carbapenem resistance in
early stages, it is imperative to develop rapid diagnostic tests
that will substantially reduce the time and cost in determining
carbapenem resistance. Thus, we devised a staining-based
diagnostic method applicable to three different Gram-negative
pathogens of Acinetobacter baumannii, Escherichia coli,
and Klebsiella pneumoniae, all with the high potential to develop
carbapenem resistance. Regardless of the resistance mechanisms
presented by bacterial species and strains, double
staining with propidium iodide (PI) and alamar blue (AB)
identified resistant bacteria with an average sensitivity of
95.35%, 7 h after imipenem treatments in 343 clinical isolates.
Among the three species tested, A. baumannii showed the
highest diagnostic sensitivity of 98.46%. The PI and ABmediated
staining method could be a promising diagnostic
method with high-throughput efficacy and low cost.
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- ppGpp-Mediated Stationary Phase Induction of the Genes Encoded by Horizontally Acquired Pathogenicity Islands and cob/pdu Locus in Salmonella enterica serovar Typhimurium
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Miryoung Song , Hyun-Ju Kim , Sangryeol Ryu , Hyunjin Yoon , Jiae Yun , Hyon E. Choy
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J. Microbiol. 2010;48(1):89-95. Published online March 11, 2010
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DOI: https://doi.org/10.1007/s12275-009-0179-6
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Salmonella enterica is highly diverse in terms of genome structure, which is at least partly due to the horizontal transfer of genetic elements from various sources. In this study, we examined the expression profiles of such genes in Salmonella Pathogenicity Islands (SPIs) and the cob/pdu locus, horizontally acquired large DNA segments, during growth under standard growth conditions. Transcripts from exponentially growing and early stationary phase Salmonellae were compared using various methods including cDNA microarray analysis. Nearly all genes encoded by SPIs and the cob/pdu locus were induced at the onset of the stationary phase in a stringent molecule ppGpp-dependent but stationary phase σ, σ38-independent manner. Although, it has been suggested that ppGpp acts in concert with DksA, we found the stationary phase induction of those SPI genes was not DksA dependent. It is suggested that ppGpp stimulates the expression of these stress-inducible genes encoded by horizontally acquired DNA, by itself or in concert with DksA.
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