Detection of point mutations associated with antibiotic resistance in Pseudomonas aeruginosa

Neda Gorgani, Scott Ahlbrand, Andrew J Patterson, Nader Pourmand

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Excessive use of broad-spectrum antibiotics in hospitals has led to the emergence of highly resistant strains of Pseudomonas aeruginosa. To reduce the selection pressure for resistance, it is important to determine the antibiotic susceptibility pattern of bacteria so that hospital patients can be treated with more narrow-spectrum and target-specific antibiotics. This study describes the development of a technique for detecting point muations in the fluoroquinolone resistance-determining region of the gyrA and parC genes as well as the efflux regulatory genes mexR, mexZ and mexOZ that are associated with fluoroquinolone and aminoglycoside resistance. The assay is based on a short DNA sequencing method using multiplex-fast polymerase chain reaction (PCR) and Pyrosequencing™ for amplification and sequencing of the selected genes. Fifty-nine clinical isolates of P. aeruginosa were examined for mutations in the abovementioned genes. Mutations related to antibiotic resistance were detected in codons 83 and 87 of gyrA and codon 126 of the mexR regulatory gene. Results of this study suggest Pyrosequencing™ as a substitute for traditional methods as it provides a rapid and reliable technique for determining the antibiotic resistance pattern of a given bacterial strain in <1 h.

Original languageEnglish (US)
Pages (from-to)414-418
Number of pages5
JournalInternational Journal of Antimicrobial Agents
Volume34
Issue number5
DOIs
StatePublished - Nov 1 2009

Fingerprint

Microbial Drug Resistance
Point Mutation
Pseudomonas aeruginosa
Fluoroquinolones
Regulator Genes
Anti-Bacterial Agents
Codon
Genes
Mutation
Multiplex Polymerase Chain Reaction
Aminoglycosides
DNA Sequence Analysis
Bacteria
Pressure

Keywords

  • Antibiotic resistance
  • Pseudomonas aeruginosa
  • Pyrosequencing

ASJC Scopus subject areas

  • Microbiology (medical)
  • Pharmacology (medical)
  • Infectious Diseases

Cite this

Detection of point mutations associated with antibiotic resistance in Pseudomonas aeruginosa. / Gorgani, Neda; Ahlbrand, Scott; Patterson, Andrew J; Pourmand, Nader.

In: International Journal of Antimicrobial Agents, Vol. 34, No. 5, 01.11.2009, p. 414-418.

Research output: Contribution to journalArticle

Gorgani, Neda ; Ahlbrand, Scott ; Patterson, Andrew J ; Pourmand, Nader. / Detection of point mutations associated with antibiotic resistance in Pseudomonas aeruginosa. In: International Journal of Antimicrobial Agents. 2009 ; Vol. 34, No. 5. pp. 414-418.
@article{2402203779aa4546b7fd36fcd268114a,
title = "Detection of point mutations associated with antibiotic resistance in Pseudomonas aeruginosa",
abstract = "Excessive use of broad-spectrum antibiotics in hospitals has led to the emergence of highly resistant strains of Pseudomonas aeruginosa. To reduce the selection pressure for resistance, it is important to determine the antibiotic susceptibility pattern of bacteria so that hospital patients can be treated with more narrow-spectrum and target-specific antibiotics. This study describes the development of a technique for detecting point muations in the fluoroquinolone resistance-determining region of the gyrA and parC genes as well as the efflux regulatory genes mexR, mexZ and mexOZ that are associated with fluoroquinolone and aminoglycoside resistance. The assay is based on a short DNA sequencing method using multiplex-fast polymerase chain reaction (PCR) and Pyrosequencing™ for amplification and sequencing of the selected genes. Fifty-nine clinical isolates of P. aeruginosa were examined for mutations in the abovementioned genes. Mutations related to antibiotic resistance were detected in codons 83 and 87 of gyrA and codon 126 of the mexR regulatory gene. Results of this study suggest Pyrosequencing™ as a substitute for traditional methods as it provides a rapid and reliable technique for determining the antibiotic resistance pattern of a given bacterial strain in <1 h.",
keywords = "Antibiotic resistance, Pseudomonas aeruginosa, Pyrosequencing",
author = "Neda Gorgani and Scott Ahlbrand and Patterson, {Andrew J} and Nader Pourmand",
year = "2009",
month = "11",
day = "1",
doi = "10.1016/j.ijantimicag.2009.05.013",
language = "English (US)",
volume = "34",
pages = "414--418",
journal = "International Journal of Antimicrobial Agents",
issn = "0924-8579",
publisher = "Elsevier",
number = "5",

}

TY - JOUR

T1 - Detection of point mutations associated with antibiotic resistance in Pseudomonas aeruginosa

AU - Gorgani, Neda

AU - Ahlbrand, Scott

AU - Patterson, Andrew J

AU - Pourmand, Nader

PY - 2009/11/1

Y1 - 2009/11/1

N2 - Excessive use of broad-spectrum antibiotics in hospitals has led to the emergence of highly resistant strains of Pseudomonas aeruginosa. To reduce the selection pressure for resistance, it is important to determine the antibiotic susceptibility pattern of bacteria so that hospital patients can be treated with more narrow-spectrum and target-specific antibiotics. This study describes the development of a technique for detecting point muations in the fluoroquinolone resistance-determining region of the gyrA and parC genes as well as the efflux regulatory genes mexR, mexZ and mexOZ that are associated with fluoroquinolone and aminoglycoside resistance. The assay is based on a short DNA sequencing method using multiplex-fast polymerase chain reaction (PCR) and Pyrosequencing™ for amplification and sequencing of the selected genes. Fifty-nine clinical isolates of P. aeruginosa were examined for mutations in the abovementioned genes. Mutations related to antibiotic resistance were detected in codons 83 and 87 of gyrA and codon 126 of the mexR regulatory gene. Results of this study suggest Pyrosequencing™ as a substitute for traditional methods as it provides a rapid and reliable technique for determining the antibiotic resistance pattern of a given bacterial strain in <1 h.

AB - Excessive use of broad-spectrum antibiotics in hospitals has led to the emergence of highly resistant strains of Pseudomonas aeruginosa. To reduce the selection pressure for resistance, it is important to determine the antibiotic susceptibility pattern of bacteria so that hospital patients can be treated with more narrow-spectrum and target-specific antibiotics. This study describes the development of a technique for detecting point muations in the fluoroquinolone resistance-determining region of the gyrA and parC genes as well as the efflux regulatory genes mexR, mexZ and mexOZ that are associated with fluoroquinolone and aminoglycoside resistance. The assay is based on a short DNA sequencing method using multiplex-fast polymerase chain reaction (PCR) and Pyrosequencing™ for amplification and sequencing of the selected genes. Fifty-nine clinical isolates of P. aeruginosa were examined for mutations in the abovementioned genes. Mutations related to antibiotic resistance were detected in codons 83 and 87 of gyrA and codon 126 of the mexR regulatory gene. Results of this study suggest Pyrosequencing™ as a substitute for traditional methods as it provides a rapid and reliable technique for determining the antibiotic resistance pattern of a given bacterial strain in <1 h.

KW - Antibiotic resistance

KW - Pseudomonas aeruginosa

KW - Pyrosequencing

UR - http://www.scopus.com/inward/record.url?scp=69949120910&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=69949120910&partnerID=8YFLogxK

U2 - 10.1016/j.ijantimicag.2009.05.013

DO - 10.1016/j.ijantimicag.2009.05.013

M3 - Article

VL - 34

SP - 414

EP - 418

JO - International Journal of Antimicrobial Agents

JF - International Journal of Antimicrobial Agents

SN - 0924-8579

IS - 5

ER -