Multiplex PCR Simultaneously Identifies N. gonorrhoeae and Predicts Ciprofloxacin Susceptibility
A SYBR green-based multiplex real-time PCR assay was developed to simultaneously detect Neisseria gonorrhoeae and determine ciprofloxacin susceptibility by targeting gyrA single nucleotide polymorphisms. Validated across panels totalling over 400 isolates, the assay achieved greater than 99% sensitivity and specificity for both pathogen identification and resistance prediction. This dual-purpose test could enable rapid point-of-care treatment decisions for gonorrhoea while supporting antimicrobial stewardship.
The original study
Multiplex Real-Time PCR Assay for Simultaneous Identification of Neisseria gonorrhoeae and Its Ciprofloxacin Susceptibility Status.
- Authors
- Perera SR, Khan NH, Martin I, Taheri A, Parti RP, Levett PN, et al.
- Journal
- Journal of clinical microbiology
- Type
- Journal Article, Validation Study
- PMID
- 28814585
Original abstract
A real-time PCR (RT-PCR) assay was designed for the simultaneous identification of Neisseria gonorrhoeae and its ciprofloxacin susceptibility status. A SYBR green-based multiplex RT-PCR format was used; it comprised two different forward primers and a common reverse primer to detect single nucleotide polymorphisms (SNPs) in gyrA of N. gonorrhoeae The primer pairs were evaluated for their sensitivity and specificity using genomic DNA from 254 N. gonorrhoeae isolates (82 were ciprofloxacin susceptible and 172 were ciprofloxacin resistant) and 23 non-N. gonorrhoeae species isolates. The performance of the primers was validated using genomic DNA from 100 different N. gonorrhoeae isolates (46 were ciprofloxacin susceptible and 54 were ciprofloxacin resistant) and 52 non-N. gonorrhoeae isolates. The latter panel was revalidated by testing 99 (46 isolates were ciprofloxacin susceptible and 53 isolates were ciprofloxacin resistant) of the N. gonorrhoeae isolates and 23 non-N. gonorrhoeae isolates. These primers detected N. gonorrhoeae and its ciprofloxacin susceptibility status with over 99% sensitivity and specificity for all panels tested. This assay has the potential to be an inexpensive and rapid test for the simultaneous identification of N. gonorrhoeae and its ciprofloxacin susceptibility status.