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Journal of Clinical Microbiology, November 2000, p. 3971-3978, Vol. 38, No. 11
National Food Safety and Toxicology
Center,1 Department of Food Science and
Human Nutrition,4 Department of
Microbiology,2 and DOE Plant Research
Laboratory,3 Michigan State University, East
Lansing, Michigan
Received 23 March 2000/Returned for modification 27 May
2000/Accepted 8 August 2000
Fluoroquinolones are one class of antimicrobial agents commonly
used to treat severe Campylobacter jejuni infection.
C. jejuni strains resistant to high levels of the
fluoroquinolone ciprofloxacin (MIC
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Identification of Ciprofloxacin-Resistant
Campylobacter jejuni by Use of a Fluorogenic PCR
Assay
16 µg/ml) have been
predominantly characterized with a C
T transition in codon 86 of
gyrA. The gyrA gene encodes one subunit of
DNA gyrase, which is a primary target for fluoroquinolone antibiotics. This study establishes a rapid PCR-based TaqMan method for identifying ciprofloxacin-resistant C. jejuni strains that carry the
C
T transition in codon 86 of gyrA. The assay
uses real-time detection, eliminating the need for gel
electrophoresis. Optimization of the assay parameters using
purified Campylobacter DNA resulted in the ability to
detect femtogram levels of DNA. The method should be useful for
monitoring the development of ciprofloxacin resistance in C. jejuni. Compiled nucleotide sequence data on the quinolone
resistance-determining region of gyrA in
Campylobacter indicate that sequence comparison of this region is a useful method for tentative identification of
Campylobacter isolates at the species level.
*
Corresponding author. Mailing address: National Food
Safety and Toxicology Center, Michigan State University, East
Lansing, MI 48824. Phone: (517) 353-9624. Fax: (517) 432-2310. E-mail: jlinz{at}msu.edu.
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