Design of a multiplex PCR assay for the simultaneous detection and confirmation of Neisseria gonorrhoeae.
AffiliationDepartment of Medical Microbiology, Cork University Hospital, Wilton, Cork, Ireland.
Polymerase Chain Reaction
Sensitivity and Specificity
MetadataShow full item record
CitationDesign of a multiplex PCR assay for the simultaneous detection and confirmation of Neisseria gonorrhoeae. 2010, 63 (5):431-3 J. Clin. Pathol.
JournalJournal of clinical pathology
AbstractTo 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.
PCR primers were designed to detect two N gonorrhoeae genes, namely porA and pgi1. Primers for an internal control targeting the ompW gene of Vibrio cholerae were also designed and incorporated in the assay. The DNA 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.
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 specificity of 100%, and a limit of detection of 1.25 ng of DNA.
This multiplex PCR assay offers a sensitive and specific assay suitable for the detection of N gonorrhoeae, and offers real potential for diagnostic use.
- A real-time PCR assay for the detection of Neisseria gonorrhoeae in genital and extragenital specimens.
- Authors: Whiley DM, Buda PP, Freeman K, Pattle NI, Bates J, Sloots TP
- Issue date: 2005 May
- Comparison of three in-house multiplex PCR assays for the detection of Neisseria gonorrhoeae and Chlamydia trachomatis using real-time and conventional detection methodologies.
- Authors: Whiley DM, Sloots TP
- Issue date: 2005 Oct
- A duplex Neisseria gonorrhoeae real-time polymerase chain reaction assay targeting the gonococcal porA pseudogene and multicopy opa genes.
- Authors: Goire N, Nissen MD, LeCornec GM, Sloots TP, Whiley DM
- Issue date: 2008 May
- Validation of a laboratory-developed real-time PCR protocol for detection of Chlamydia trachomatis and Neisseria gonorrhoeae in urine.
- Authors: Hopkins MJ, Ashton LJ, Alloba F, Alawattegama A, Hart IJ
- Issue date: 2010 Jun
- Clinical validation of a real-time polymerase chain reaction detection of Neisseria gonorrheae porA pseudogene versus culture techniques.
- Authors: Hjelmevoll SO, Olsen ME, Sollid JU, Haaheim H, Melby KK, Moi H, Unemo M, Skogen V
- Issue date: 2008 May