379 ISSN 0026-8933, Molecular Biology, 2017, Vol. 51, No. 3, pp. 379–388. © Pleiades Publishing, Inc., 2017. Original Russian Text © A.A. Kubanov, A.T. Leinsoo, A.V. Chestkov, E.I. Dementieva, B.L. Shaskolskiy, V.S. Solomka, D.A. Gryadunov, D.G. Deryabin, 2017, published in Molekulyarnaya Biologiya, 2017, Vol. 51, No. 3, pp. 431–441. Drug Resistance Mutations and Susceptibility Phenotypes of Neisseria gonorrhoeae Isolates in Russia A. A. Kubanov a , A. T. Leinsoo b , A. V. Chestkov a , E. I. Dementieva b , B. L. Shaskolskiy b , V. S. Solomka a , D. A. Gryadunov b , and D. G. Deryabin a, * a State Research Center of Dermatovenerology and Cosmetology, Ministry of Health of Russian Federation, Moscow, 107076 Russia b Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 199991 Russia *e-mail: dgderyabin@yandex.ru Received March 30, 2016; in final form, June 14, 2016 AbstractSteady growth in the degree of antimicrobial resistance in Neisseria gonorrhoeae calls for the con- trol of the spreading of resistance mutations. Here we present the data describing drug resistance mutations, the results of antimicrobial susceptibility tests, and molecular genotypes of 128 recent N. gonorrhoeae isolates collected across 9 regions of the Russian Federation. The mutations in chromosome genes penA, ponA, rpsJ, gyrA, parC, which determine the susceptibility of N. gonorrhoeae to penicillins, tetracyclines, and fluoro- quinolones were detected by multiplex amplification followed by hybridization on a hydrogel microarray. The most frequent mutation was an insertion of an aspartate at position 345 of penA gene (76.6%), whereas muta- tions Leu421Pro in ponA gene, Val57Met in rpsJ gene, Ser91Phe in gyrA gene, Asp95Gly in gyrA gene, and Ser87Arg in parC gene were detected in 32.8–36.7% of strains. One third of studied N. gonorrhoeae isolates har- bored multiple drug resistance mutations in bacterial chromosome, resulting in the bimodal distribution of muta- tion profiles and related patterns of antimicrobial susceptibility. The spread of multiple resistance could be explained by the vertical transfer of the mutations resulting in the clonality of the N. gonorrhoeae population. Keywords: Neisseria gonorrhoeae, genetic determinants of drug resistance, phenotypic susceptibility, DNA microarray, statistical analysis, multiple drug resistance DOI: 10.1134/S0026893317030116 INTRODUCTION The emergence and rapid spread of microorganism resistance to antibiotics introduced into clinical prac- tice is a global problem in medicine [1]. The danger of the development of total resistance to antibacterial agents for a number of agents causing infectious dis- eases has been registered in WHO reports [2, 3]. Microorganisms of Neisseria gonorrhoeae species are of special concern since their progressive resistance creates the prospect for gonorrhea to transform into the category of a potentially incurable infection [4]. Known pathways of developing drug resistance are associated with two main mechanisms, i.e., (1) acquisi- tion of new genes that are frequently exposed to horizon- tal transfer within mobile elements (plasmids and trans- posons) and encode enzymes that inactivate drug mole- cules; (2) changes in the chromosomal genes that encode targets for their effect [5]. The analysis of drug resistance of gonococci [6] indicates the prevailing significance of the second mechanism associated with changes of their own genome. In particular, the N. gonorrhoeae resis- tance to penicillin caused by the presence of TEM-1- like β-lactamase is not widespread, while mutations in chromosomal ponA (Leu to Pro amino acid substitu- tion in the position 421) [7] and penA (insertion of three nucleotides with the appearance of Asp amino acid in the 345 position) [8] genes resulted in a signif- icant decrease in the affinity of penicillin-binding pro- teins encoded by these genes with a loss of therapeutic efficiency of penicillin during the treatment of gono- coccal infection in the 1970s. The resistance of N. gon- orrhoeae to tetracycline (developed at the same time) is associated with a point mutation in the rpsJ gene that encodes the ribosomal S10 protein [9] amplified by a combination of mutations in the mtrR genes (the repressor of mtrCDE operon transcription responsible for the regulation of the expression of the MtrC– MtrD–MtrE, efflux pump, which removes the drug from periplasmic space of gonococci), and penB gene (the porin protein, which provides the drug transport through the external membrane) [10]. Fluoroquinolo- nes, which were used to treat gonococcus infections in the 1980s, also maintained their efficiency for only a short time. Reports about failures in their use already began to appear 10 years later [11] and this group of Abbreviations: MIC, minimal inhibiting concentration. GENOMICS. TRANSCRIPTOMICS UDC 57.088.1,579.611