A Multiplex Molecular Assay for Detection of Six penA Codons To Predict Decreased Susceptibility to Cephalosporins in Neisseria gonorrhoeae.
Yamei LiLulu ZhangLeshan XiuDi WangYaling ZengFeng WangYueping YinJunping PengPublished in: Antimicrobial agents and chemotherapy (2022)
The emerging cephalosporin-resistant Neisseria gonorrhoeae poses an urgent threat to the continued efficacy of the last-line monotherapy for gonorrhea. Consequently, high-throughput, accurate, and reasonable molecular assays are urgently needed for strengthening antimicrobial-resistance surveillance in N. gonorrhoeae. In this study, we designed a high-throughput multiplex method that incorporates high-resolution melting technology and is based on a 6-codon assay (among the most parsimonious assays) developed following comprehensive and systematic reviews. The results showed that our method can precisely distinguish specific single-nucleotide polymorphisms in resistance-associated genes with a specificity and sensitivity of 100% and a detection limit as low as 10 copies per reaction. This method can be directly applied to clinical samples without cumbersome culture and successfully predicted all cephalosporin-resistant isolates (sensitivity: 100%). The method presented here represents a technique for rapid testing of antimicrobial resistance and will serve as a valuable tool for tailor-made antimicrobial therapy and for monitoring the transmission of cephalosporin-resistant strains.
Keyphrases
- high throughput
- antimicrobial resistance
- high resolution
- single cell
- loop mediated isothermal amplification
- real time pcr
- systematic review
- public health
- escherichia coli
- gram negative
- staphylococcus aureus
- mass spectrometry
- stem cells
- randomized controlled trial
- genome wide
- gene expression
- bone marrow
- clinical trial
- label free
- multidrug resistant
- study protocol
- meta analyses
- replacement therapy