Design of a multiplex PCR assay for the simultaneous detection and confirmation of Neisseria gonorrhoeae

Objectives To improve the detection of Neisseria gonorrhoeae by designing a multiplex PCR assay using two N gonorrhoeae-specific genes as targets, thereby providing detection and confirmation of a positive result simultaneously. Methods PCR primers were designed to detect two N gonorrhoeae genes, namely porA and pgi1. Primers for an internal controltargeting the ompW gene of Vibrio cholerae werealsodesignedandincorporatedintheassay.TheDNA of 45 clinical isolates including 33 N gonorrhoeae isolates, seven non-gonococcal Neisseria species, and five non-Neisseria species was tested using the multiplex PCR assay. Results All 33 N gonorrhoeae isolates were successfully detected by the assay and none of the non-gonococcal isolates was detected. The assay showed a sensitivity and specificityof100%,andalimitofdetectionof1.25 ngofDNA. Conclusion This multiplex PCR assay offers a sensitive and specific assay suitable for the detection of N gonorrhoeae, and offers real potential for diagnostic use.