Journal of Clinical Microbiology
Journal of Clinical Microbiology
5 years ago

A Multiplex RT-PCR Assay for the Simultaneous Identification of Neisseria gonorrhoeae and Its Ciprofloxacin Susceptibility Status.

Perera SR, Parti RP, Dillon JR, Kusalik A, Horsman GB, Martin I, Khan NH, Levett PN, Taheri A
A real time PCR (RT-PCR) assay was designed for the simultaneous identification of N. gonorrhoeae and its ciprofloxacin susceptibility status. A SYBR-Green based multiplex RT-PCR format was used and 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 with genomic DNA from 254 N. gonorrhoeae isolates (82 were ciprofloxacin susceptible and 172 were ciprofloxacin resistant) and 23 non-N. gonorrhoeae species. 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. This 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. his assay has the potential to be an inexpensive and rapid test for the simultaneous identification of N. gonorrhoeae and its ciprofloxacin susceptibility status.

Publisher URL: https://www.ncbi.nlm.nih.gov/pubmed/28814585

DOI: PubMed:28814585

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