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

Liben Chen, Dong Jin Shin, Shuyu Zheng, Johan H. Melendez, Charlotte A. Gaydos, Tza Huei Wang

Research output: Contribution to journalArticle

Abstract

Multidrug-resistant gonorrhea has become an urgent issue for global public health. As the causative agent of gonorrhea, Neisseria gonorrhoeae, has been progressively developing resistance to nearly all prescribed antimicrobial drugs, monitoring its antimicrobial resistance on a broader scale has become a crucial agenda for effective antibiotic stewardship. Unfortunately, gold standard antimicrobial susceptibility testing (AST) relies on time and labor-intensive phenotypic assays, which lag behind the current diagnostic workflow for N. gonorrhoeae identification based on nucleic acid amplification tests (NAAT). Newer assay technologies based on NAAT can rapidly identify N. gonorrhoeae from clinical specimen but fundamentally lack the capacity to provide phenotypic AST information. Herein, we propose a direct-quantitative PCR (direct-qPCR) assay that enables pathogen-specific identification and phenotypic AST via quantitative measurement of N. gonorrhoeae growth directly from a liquid medium without any sample preprocessing. The assay has an analytical sensitivity of 10 2 CFU/mL and is highly specific to N. gonorrhoeae in the presence of urogenital flora and clinical swab eluent. We tested seven N. gonorrhoeae strains against three antibiotic agents, penicillin, tetracycline, and ciprofloxacin, and achieved 95.2% category agreement and 85.7% essential agreement with the FDA-approved E-test. The assay presented in this work has the unique ability to identify N. gonorrhoeae and provide phenotypic AST directly from the liquid medium with cell densities as low as 10 2 CFU/mL, demonstrating an accelerated, sensitive, and scalable workflow for performing both identification and AST of N. gonorrhoeae.

Original languageEnglish (US)
Pages (from-to)1377-1384
Number of pages8
JournalACS Infectious Diseases
Volume4
Issue number9
DOIs
StatePublished - Jul 12 2018

Keywords

  • Neisseria gonorrhoeae
  • antimicrobial resistance
  • ciprofloxacin
  • direct PCR
  • growth measurement
  • pathogen identification
  • penicillin
  • phenotypic antimicrobial susceptibility testing
  • tetracycline

ASJC Scopus subject areas

  • Infectious Diseases

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