title: Multiplex bead suspension array for screening Neisseria gonorrhoeae antibiotic resistance genetic determinants in noncultured clinical samples.
abstract: The increasing threat of antibiotic-resistant Neisseria gonorrhoeae highlights the need for new diagnostic options. A high-throughput multiplex bead suspension array assay was developed for profiling 29 N. gonorrhoeae genomic mutations and 2 plasmid genes conferring resistance to 6 antimicrobial agents: penicillin, ciprofloxacin, cefixime, tetracycline, azithromycin, and spectinomycin. The three steps of this assay include amplification of 12 N. gonorrhoeae chromosomal and plasmid loci, multiplex allele-specific primer extension reaction, and multiplex bead suspension array detection. Antibiotic resistance genetic determinants were identified successfully in 239 cervicovaginal N. gonorrhoeae-positive noncultured swab samples. This molecular assay can be used for detection of gonococci in clinical specimens, molecular typing, mutation profiling, and predictive assessment of N. gonorrhoeae susceptibility to antibiotics without the need for culture.
authors.affiliation: Femeris Women's Health Research Center, Medical Diagnostic Laboratories, LLC, a Member of the Genesis Biotechnology Group, Hamilton, New Jersey 08690, USA. sbalashov@mdlab.com
authors.forename: Sergey
authors.initials: S
authors.lastname: Balashov
authors.forename: Eli
authors.initials: E
authors.lastname: Mordechai
authors.forename: Martin E
authors.initials: ME
authors.lastname: Adelson
authors.forename: Scott E
authors.initials: SE
authors.lastname: Gygax
mesh_terms: D000483:Alleles; D000890:Anti-Infective Agents / Q000494:pharmacology; D017963:Azithromycin / Q000494:pharmacology; D020682:Cefixime / Q000494:pharmacology; D002511:Cephalosporins / Q000494:pharmacology; D002876:Chromosomes, Bacterial / Q000235:genetics; D002939:Ciprofloxacin / Q000494:pharmacology; D004269:DNA, Bacterial / Q000235:genetics; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D020869:Gene Expression Profiling; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D008826:Microbial Sensitivity Tests; D009154:Mutation; D009344:Neisseria gonorrhoeae / Q000187:drug effects* / Q000235:genetics* / Q000254:growth & development; D010406:Penicillins / Q000494:pharmacology; D010957:Plasmids; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods; D000198:Spectinomycin / Q000494:pharmacology; D013752:Tetracycline / Q000494:pharmacology
chemical_list: D000890:Anti-Infective Agents; D002511:Cephalosporins; D004269:DNA, Bacterial; D010406:Penicillins; D002939:Ciprofloxacin; D017963:Azithromycin; D000198:Spectinomycin; D020682:Cefixime; D013752:Tetracycline