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Antimicrobial Agents and Chemotherapy, January 2001, p. 117-123, Vol. 45, No. 1
0066-4804/01/$04.00+0   DOI: 10.1128/AAC.45.1.117-123.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Molecular Basis of High-Level Ciprofloxacin Resistance in Neisseria gonorrhoeae Strains Isolated in Denmark from 1995 to 1998

Xiaohong Sudagger and Inga Lind*

Neisseria Unit, Department of Respiratory Infections, Meningitis and STIs, Statens Serum Institut, Copenhagen, Denmark

Received 14 April 2000/Returned for modification 12 July 2000/Accepted 3 October 2000

In Denmark surveillance of the in vitro susceptibility to ciprofloxacin of Neisseria gonorrhoeae was established in 1990. The proportion of N. gonorrhoeae strains with decreased susceptibility or resistance to ciprofloxacin (MIC >=  0.06 µg/ml) was low (0.3 to 2.3%) up to 1995. Between 1995 and 1998 the rate of less-susceptible and resistant strains rose from 6.9 to 13.2%. Among ciprofloxacin-resistant strains (MIC >=  1 µg/ml), 81% were highly resistant (MIC >=  4 µg/ml). Thirty-five N. gonorrhoeae strains (40 isolates) for which ciprofloxacin MICs were 4 to 32 µg/ml were investigated for the frequency and patterns of mutations within the gyrA and parC genes. The quinolone resistance-determining regions of the gyrA and parC genes were amplified by PCR, and the amplicons were directly sequenced. Alterations at Ser-91 and Asp-95 in GyrA and a single or double alteration in ParC were identified in 32 strains (91%). Ser-91-to-Phe and Asp-95-to-Gly alterations in GyrA were detected in 28 strains (80%). The most common ParC alteration, Asp-86 to Asn, was found in 19 strains (54%). The strains were analyzed for genetic relationship by pulsed-field gel electrophoresis (PFGE). The analysis showed that nine strains with the same mutation pattern in the gyrA and parC genes, originating from different geographical areas over 3 years, had the same PFGE patterns after SpeI as well as NheI digestion (only one strain with one band difference in the NheI pattern), suggesting that a resistant clone had spread worldwide. The results from this study strongly suggest that double gyrA mutations plus a parC mutation(s) play an important role in the development of high-level fluoroquinolone resistance in N. gonorrhoeae.

* Corresponding author. Mailing address: Neisseria Unit, Department of Respiratory Infections, Meningitis and STIs, Statens Serum Institut, Artillerivej 5, 2300 Copenhagen S, Denmark. Phone: (45) 3268 3475. Fax: (45) 32683142. E-mail: il{at}ssi.dk.

dagger Present address: Department of STD, National Center for STD and Leprosy Control, Institute of Dermatology, Chinese Academy of Medical Sciences, Nanjing 210042, People's Republic of China.

Antimicrobial Agents and Chemotherapy, January 2001, p. 117-123, Vol. 45, No. 1
0066-4804/01/$04.00+0   DOI: 10.1128/AAC.45.1.117-123.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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