Title : Molecular drug susceptibility testing and genotyping of Neisseria gonorrhoeae isolates in Russia
Gonococcal infection belongs to the most common sexually transmitted infections. Due to the developing antimicrobial resistance, in particular, resistance to fluoroquinolones and 3rd generation cephalosporins, Neisseria gonorrhoeae is considered to be a high priority pathogen according to the WHO list.
In this work, we analyzed 270 N. gonorrhoeae clinical isolates collected in Russia in 2018–2019. The phenotypic susceptibility testing was performed using agar dilution method. Hydrogel microarray with immobilized oligonucleotide probes was developed for the identification of genetic markers causing the N. gonorrhoeae resistance to penicillins, fluoroquinolones, and cephalosporins: mutations in the penA gene encoding penicillin-binding protein PBP2 (Ala311Val, Ile312Met, Val316Thr/Pro, ins345Asp, Thr483Ser, Ala501Val/Pro/Thr, Asn512Tyr, Gly545Ser, Pro551Ser/Leu), ponA gene (Leu421Pro), mtrR promoter region (-35delA, -10insT), porB gene (Gly120Lys/Asp/Asn/Thr, Ala121Ala/Asp/Asn/Gly), gyrA gene (Ser91Phe/Thr, Asp95Ala/Gly), and parC gene (Ser87Arg/Asn/Ile, Glu 91Lys). The mutations in the penA gene, which are analyzed using microarray, can identify the presence of the mosaic penA alleles and alterations associated with resistance to 3rd generation cephalosporins. For validation microarray results and genotyping (N. gonorrhoeae multiantigen sequence typing, NG-MAST) Sanger sequencing was performed.