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

Hdl Handle:
http://hdl.handle.net/10147/200274
Title:
Design of a multiplex PCR assay for the simultaneous detection and confirmation of Neisseria gonorrhoeae.
Authors:
O'Callaghan, Isabelle; Corcoran, Daniel; Lucey, Brigid
Affiliation:
Department of Medical Microbiology, Cork University Hospital, Wilton, Cork, Ireland.
Citation:
Design of a multiplex PCR assay for the simultaneous detection and confirmation of Neisseria gonorrhoeae. 2010, 63 (5):431-3 J. Clin. Pathol.
Journal:
Journal of clinical pathology
Issue Date:
May-2010
URI:
http://hdl.handle.net/10147/200274
DOI:
10.1136/jcp.2009.071431
PubMed ID:
20360139
Abstract:
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.; 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.
Item Type:
Article
Language:
en
Description:
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 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. 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 specificity of 100%, and a limit of detection of 1.25 ng of DNA. 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.
MeSH:
DNA, Bacterial; Genes, Bacterial; Gonorrhea; Humans; Neisseria gonorrhoeae; Polymerase Chain Reaction; Porins; Sensitivity and Specificity
ISSN:
1472-4146

Full metadata record

DC FieldValue Language
dc.contributor.authorO'Callaghan, Isabelleen
dc.contributor.authorCorcoran, Danielen
dc.contributor.authorLucey, Brigiden
dc.date.accessioned2012-01-05T14:09:20Z-
dc.date.available2012-01-05T14:09:20Z-
dc.date.issued2010-05-
dc.identifier.citationDesign of a multiplex PCR assay for the simultaneous detection and confirmation of Neisseria gonorrhoeae. 2010, 63 (5):431-3 J. Clin. Pathol.en
dc.identifier.issn1472-4146-
dc.identifier.pmid20360139-
dc.identifier.doi10.1136/jcp.2009.071431-
dc.identifier.urihttp://hdl.handle.net/10147/200274-
dc.descriptionOBJECTIVES: 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 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. 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 specificity of 100%, and a limit of detection of 1.25 ng of DNA. 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.en
dc.description.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.-
dc.description.abstractPCR 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.-
dc.description.abstractAll 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.-
dc.description.abstractThis multiplex PCR assay offers a sensitive and specific assay suitable for the detection of N gonorrhoeae, and offers real potential for diagnostic use.-
dc.language.isoenen
dc.subject.meshDNA, Bacterial-
dc.subject.meshGenes, Bacterial-
dc.subject.meshGonorrhea-
dc.subject.meshHumans-
dc.subject.meshNeisseria gonorrhoeae-
dc.subject.meshPolymerase Chain Reaction-
dc.subject.meshPorins-
dc.subject.meshSensitivity and Specificity-
dc.titleDesign of a multiplex PCR assay for the simultaneous detection and confirmation of Neisseria gonorrhoeae.en
dc.typeArticleen
dc.contributor.departmentDepartment of Medical Microbiology, Cork University Hospital, Wilton, Cork, Ireland.en
dc.identifier.journalJournal of clinical pathologyen
dc.description.provinceMunster-
All Items in Lenus, The Irish Health Repository are protected by copyright, with all rights reserved, unless otherwise indicated.