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Antimicrobial Agents and Chemotherapy, June 2008, p. 2175-2182, Vol. 52, No. 6
0066-4804/08/$08.00+0     doi:10.1128/AAC.01420-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Relation between Genetic Markers of Drug Resistance and Susceptibility Profile of Clinical Neisseria gonorrhoeae Strains{triangledown}

Elena N. Ilina,1* Vladimir A. Vereshchagin,1 Alexandra D. Borovskaya,1 Maja V. Malakhova,1 Sergei V. Sidorenko,2,3 Nazar C. Al-Khafaji,2 Anna A. Kubanova,2 and Vadim M. Govorun1

Research Institute of Physico-Chemical Medicine, Moscow, Russian Federation,1 Central Research Institute of Dermatology and Venerology, Moscow, Russian Federation,2 National Research Centre for Antibiotics, Moscow, Russian Federation3

Received 2 November 2007/ Returned for modification 15 January 2008/ Accepted 20 March 2008

The main goal of this work is to clarify the predictive value of known genetic markers of Neisseria gonorrhoeae resistance to penicillin, tetracycline, and fluoroquinolones. The correlation between the presence of certain genetic markers and susceptibility of N. gonorrhoeae isolates to penicillin, tetracycline, and fluoroquinolones has been analyzed by means of statistical methods. Susceptibility testing with penicillin, tetracycline, and fluoroquinolones was performed by the agar dilution method. N. gonorrhoeae genomic DNA was isolated. The presence of blaTEM-1 and tet(M) genes was analyzed by PCR. A novel method of polymorphism discovery based on a minisequencing reaction followed by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry was applied for the analysis of chromosomal N. gonorrhoeae genes involved in antimicrobial resistance development. Clinical N. gonorrhoeae isolates (n = 464) were collected. Susceptibility levels to penicillin, tetracycline, and fluoroquinolones were found to be 25.9%, 35.9%, and 54.1%, respectively. Among the 19 N. gonorrhoeae isolates with penicillin MICs of ≥4 µg/ml, the blaTEM-1 gene was detected in 12. The Tet(M) determinant was found in 4 of 12 N. gonorrhoeae isolates with tetracycline MICs of ≥16 µg/ml. The chromosomal genetic markers of penicillin and tetracycline resistance were detected especially in isolates with penicillin MICs of 0.25 to 2.0 µg/ml and tetracycline MICs of 0.5 to 4 µg/ml. Mutations in GyrA and ParC were found in 208 of 211 quinolone-resistant N. gonorrhoeae isolates. This work is the first representative molecular research of the N. gonorrhoeae population in Russia. Information about the prevalence of antibiotic resistance mechanisms and the positive predictive value of certain genetic determinants is given. The positive predictive values of the analyzed genetic markers were found to be different for fluoroquinolones (90.3%), penicillin (91.1%), and tetracycline (81.9%).

* Corresponding author. Mailing address: Institute of Physico-Chemical Medicine, Malaya Pirogovskaya St., 1a, 119992 Moscow, Russia. Phone: 7-495-2450471. Fax: 7-495-2467721. E-mail: ilinaEN{at}gmail.com

{triangledown} Published ahead of print on 31 March 2008.

Antimicrobial Agents and Chemotherapy, June 2008, p. 2175-2182, Vol. 52, No. 6
0066-4804/08/$08.00+0     doi:10.1128/AAC.01420-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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