Direct-qPCR Assay for Coupled Identification and Antimicrobial Susceptibility Testing of Neisseria gonorrhoeae

• DOI: 10.1021/acsinfecdis.8b00104
• Publication Date: 2018-07-12T13:15:07Z
• Authors: Liben Chen; Dong Jin Shin; Shuyu Zheng; Johan H. Melendez; Charlotte A. Gaydos; Tza-Huei Wang
• Keywords: Neisseria gonorrhoeae

Multidrug-resistant gonorrhea has become an urgent issue for global public health. As the causative agent of gonorrhea, Neisseria gonorrhoeae, been progressively developing resistance to nearly all prescribed antimicrobial drugs, monitoring its on a broader scale crucial agenda effective antibiotic stewardship. Unfortunately, gold standard susceptibility testing (AST) relies time and labor-intensive phenotypic assays, which lag behind current diagnostic workflow N. gonorrhoeae identification based nucleic acid amplification tests (NAAT). Newer assay technologies NAAT can rapidly identify from clinical specimen but fundamentally lack capacity provide AST information. Herein, we propose direct-quantitative PCR (direct-qPCR) that enables pathogen-specific via quantitative measurement growth directly liquid medium without any sample preprocessing. The analytical sensitivity 102 CFU/mL is highly specific in presence urogenital flora swab eluent. We tested seven strains against three agents, penicillin, tetracycline, ciprofloxacin, achieved 95.2% category agreement 85.7% essential with FDA-approved E-test. presented this work unique ability cell densities as low CFU/mL, demonstrating accelerated, sensitive, scalable performing both gonorrhoeae.