This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Hurtle, W. Articles by Norwood, D. Search for Related Content PubMed PubMed Citation Articles by Hurtle, W. Articles by Norwood, D.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, July 2003, p. 3273-3283, Vol. 41, No. 7
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.7.3273-3283.2003

Detection and Identification of Ciprofloxacin-Resistant Yersinia pestis by Denaturing High-Performance Liquid Chromatography

William Hurtle,¹ Luther Lindler,² Wei Fan,² David Shoemaker,³ Erik Henchal,³ and David Norwood^3*

Clinical Research Management, North Royalton, Ohio,¹ Walter Reed Army Institute of Research, Silver Spring, Maryland,² U.S. Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland³

Received 25 September 2002/ Returned for modification 19 December 2002/ Accepted 27 March 2003

Denaturing high-performance liquid chromatography (DHPLC) has been used extensively to detect genetic variation. We used this method to detect and identify Yersinia pestis KIM5 ciprofloxacin-resistant isolates by analyzing the quinolone resistance-determining region (QRDR) of the gyrase A gene. Sequencing of the Y. pestis KIM5 strain gyrA QRDR from 55 ciprofloxacin-resistant isolates revealed five mutation types. We analyzed the gyrA QRDR by DHPLC to assess its ability to detect point mutations and to determine whether DHPLC peak profile analysis could be used as a molecular fingerprint. In addition to the five mutation types found in our ciprofloxacin-resistant isolates, several mutations in the QRDR were generated by site-directed mutagenesis and analyzed to further evaluate this method for the ability to detect QRDR mutations. Furthermore, a blind panel of 42 samples was analyzed by screening for two mutant types to evaluate the potential diagnostic value of this method. Our results showed that DHPLC is an efficient method for detecting mutations in genes that confer antibiotic resistance.

---

* Corresponding author. Mailing address: U.S. Army Medical Research Institute of Infectious Diseases, Diagnostic Systems Division, 1425 Porter St., Fort Detrick, MD 21702. Phone: (301) 619-4721. Fax: (301) 619-2492. E-mail: David.Norwood{at}det.amedd.army.mil.

---

Journal of Clinical Microbiology, July 2003, p. 3273-3283, Vol. 41, No. 7
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.7.3273-3283.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

This article has been cited by other articles:

• Arlen, P. A., Singleton, M., Adamovicz, J. J., Ding, Y., Davoodi-Semiromi, A., Daniell, H. (2008). Effective Plague Vaccination via Oral Delivery of Plant Cells Expressing F1-V Antigens in Chloroplasts. Infect. Immun. 76: 3640-3650 [Abstract] [Full Text]  
• Xu, L., Evans, J., Ling, T., Nye, K., Hawkey, P. (2007). Rapid Genotyping of CTX-M Extended-Spectrum {beta}-Lactamases by Denaturing High-Performance Liquid Chromatography. Antimicrob. Agents Chemother. 51: 1446-1454 [Abstract] [Full Text]  
• Anisimov, A. P., Amoako, K. K. (2006). Treatment of plague: promising alternatives to antibiotics.. J Med Microbiol 55: 1461-1475 [Abstract] [Full Text]  
• Woodford, N., Sundsfjord, A. (2005). Molecular detection of antibiotic resistance: when and where?. J Antimicrob Chemother 56: 259-261 [Abstract] [Full Text]