ABSTRACT
We developed a multiplexed DNA microarray-based assay allowing identification of 12 causative agents of reproductive tract infections with the simultaneous detection of 47 genetic determinants of resistance to antimicrobial substances. The microarray was tested on 93 isolates of Neisseria gonorrhoeae, 32 isolates of Treponema pallidum and 29 samples of Ureaplasma spp./Mycoplasma spp. The N. gonorrhoeae isolates had multiple mutations in the penA, ponA, rpsJ, gyrA, parC, and mtrR genes; their prognostic value significantly increased when combinations of mutations were detected. In the analyzed T. pallidum isolates, single A2058G substitution in the 23S rRNA gene responsible for macrolide resistance was found. DNA sequences of Ureaplasma spp./Mycoplasma spp. were determined as wild type, which was not fully consistent with the results of analysis of their antimicrobial susceptibility.
Subject(s)
Anti-Bacterial Agents/pharmacology , Gonorrhea/diagnosis , Molecular Diagnostic Techniques , Syphilis/diagnosis , Ureaplasma Infections/diagnosis , DNA, Bacterial/genetics , Drug Resistance, Bacterial , Gonorrhea/microbiology , Humans , Microbial Sensitivity Tests , Microchip Analytical Procedures , Neisseria gonorrhoeae/genetics , Neisseria gonorrhoeae/isolation & purification , Nucleic Acid Hybridization , Syphilis/microbiology , Treponema pallidum/genetics , Treponema pallidum/isolation & purification , Ureaplasma/genetics , Ureaplasma/isolation & purification , Ureaplasma Infections/microbiologyABSTRACT
9alpha-Hydroxy derivatives were prepared from 11 steroids ofandrostane and pregnane series using Rhodococcus erythropolis VKPM Ac-1740 culture with 0.5-20 g/l substrate concentration in the reaction mixture. 9alpha-Monohydroxylation proceeded regardless of the substituent structure at C17. However, the structure of the steroid molecule influenced the time of complete conversion of the substrate and the yield of the transformation product. 9alpha-Hydroxy-androstenedione was obtained in 35 h in a yield of 85% when the maximum concentration of androstenedione (AD) was 10 g/l. 9alpha-Hydroxy-AD was also formed by the actinobacterium cells entrapped in poly(vinyl alcohol) cryogel beads. Nine successive transformation cycles were carried out using immobilized cells at 4.0 g/l concentration of AD in the medium. The yield of 9alpha-hydroxy-AD formed during six cycles (from two to eight with the duration of each cycle for 22-24 h) was 98%.
