ArticleJournal of clinical microbiology2001
Detection of ciprofloxacin-resistant Yersinia pestis by fluorogenic PCR using the LightCycler.
Article in Journal of clinical microbiology, 2001. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed, 52 citations in OpenAlex.
- Multiplex PCR System for the Diagnosis of Plague.Current molecular medicine · 2025Review
- Applications of polymerase chain reaction-based methods for the diagnosis of plague (Review).Experimental and therapeutic medicine · 2022Review
- Beating the Bio-Terror Threat with Rapid Antimicrobial Susceptibility Testing.Microorganisms · 2021Review
- Computer-Aided Rational Engineering of Signal Sensitivity of Quorum Sensing Protein LuxR in a Whole-Cell Biosensor.Frontiers in molecular biosciences · 2021Article
- A Survey of Antimicrobial Resistance Determinants in Category A Select Agents, Exempt Strains, and Near-Neighbor Species.International journal of molecular sciences · 2020Article
- Genome-wide assessment of antimicrobial tolerance inMicrobial genomics · 2019Article
- Rapid Detection of Genetic Engineering, Structural Variation, and Antimicrobial Resistance Markers in Bacterial Biothreat Pathogens by Nanopore Sequencing.Scientific reports · 2019Article
- A Rapid Antimicrobial Susceptibility Test for DeterminingFrontiers in microbiology · 2019Article
- Targeted next-generation sequencing for the detection of ciprofloxacin resistance markers using molecular inversion probes.Scientific reports · 2016Article
- A Rapid Molecular Test for Determining Yersinia pestis Susceptibility to Ciprofloxacin by the Quantification of Differentially Expressed Marker Genes.Frontiers in microbiology · 2016Article
- Genome-level transcription data of Yersinia pestis analyzed with a new metabolic constraint-based approach.BMC systems biology · 2012Article
- Use of an in vitro pharmacodynamic model to derive a moxifloxacin regimen that optimizes kill of Yersinia pestis and prevents emergence of resistance.Antimicrobial agents and chemotherapy · 2011Article
- Ambient stable quantitative PCR reagents for the detection of Yersinia pestis.PLoS neglected tropical diseases · 2010Article
- Diagnostic bioluminescent phage for detection of Yersinia pestis.Journal of clinical microbiology · 2009Article
- Real-time PCR in clinical microbiology: applications for routine laboratory testing.Clinical microbiology reviews · 2006Review
- Real-time Fluorescent PCR Techniques to Study Microbial-Host Interactions.Methods in microbiology · 2004Review
- Ciprofloxacin susceptibility of Pseudomonas aeruginosa isolates from keratitis.The British journal of ophthalmology · 2003Article
- Real-time PCR assay for detection of fluoroquinolone resistance associated with grlA mutations in Staphylococcus aureus.Journal of clinical microbiology · 2003Article
- Detection and identification of ciprofloxacin-resistant Yersinia pestis by denaturing high-performance liquid chromatography.Journal of clinical microbiology · 2003Article
- Detection of rpoB mutations in Mycobacterium tuberculosis with LightCycler technology.Journal of clinical microbiology · 2002Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We have developed a fluorescence resonance energy transfer (FRET)-based assay to detect ciprofloxacin resistant (Cp(r)) mutants of the biothreat agent Yersinia pestis. We selected spontaneous mutants of the attenuated Y. pestis KIM 5 strain that were resistant to a ciprofloxacin (CIP) concentration of at least 1 microg/ml. DNA sequencing of gyrA encoded by 65 of these mutants revealed that all isolates contained one of four different point mutations within the quinolone resistance-determining region of gyrA. We developed a FRET-based assay that detected all of these mutations by using a single pair of fluorescent probes with sequences complementary to the wild-type Y. pestis gyrA sequence. Melting peak analysis revealed that the probe-PCR product hybrid was less stable when amplification occurred from any of the four mutant templates. This instability resulted in the PCR product obtained from the Cp(r) Y. pestis strains displaying a 4 to 11 degrees C shift in probe melting temperature. Following optimization of the reaction conditions, we were able to detect approximately 10 pg of purified wild-type template DNA or the presence of approximately 4 CFU of wild-type Y. pestis KIM 5 or Cp(r) mutants in crude lysates. Taken together, our results demonstrate the utility of FRET-based assays for detection of Cp(r) mutants of Y. pestis. This method is both sensitive and rapid.
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