A duplex Neisseria gonorrhoeae real-time polymerase chain reaction assay targeting the gonoccoccal porA pseudogene and multicopy opa genes

Goire, Namraj, Nissen, Michael D., LeCornec, Genevera M., Sloots, Theo P. and Whiley, David, M. (2008) A duplex Neisseria gonorrhoeae real-time polymerase chain reaction assay targeting the gonoccoccal porA pseudogene and multicopy opa genes. Diagnostic Microbiology and Infectious Disease, 61 1: 6-12. doi:10.1016/j.diagmicrobio.2007.12.007


Author Goire, Namraj
Nissen, Michael D.
LeCornec, Genevera M.
Sloots, Theo P.
Whiley, David, M.
Title A duplex Neisseria gonorrhoeae real-time polymerase chain reaction assay targeting the gonoccoccal porA pseudogene and multicopy opa genes
Formatted title
A duplex Neisseria gonorrhoeae real-time polymerase chain reaction assay targeting the gonoccoccal porA pseudogene and multicopy opa genes
Journal name Diagnostic Microbiology and Infectious Disease   Check publisher's open access policy
ISSN 0732-8893
1879-0070
Publication date 2008-01-01
Year available 2008
Sub-type Article (original research)
DOI 10.1016/j.diagmicrobio.2007.12.007
Open Access Status
Volume 61
Issue 1
Start page 6
End page 12
Total pages 7
Editor R. N. Jones
Place of publication Burlington, MA, U.S.A.
Publisher Elsevier
Language eng
Subject 060502 Infectious Agents
060506 Virology
060503 Microbial Genetics
920109 Infectious Diseases
Abstract Cross-reactions of gonococcal nucleic acid amplification tests (NAATs) with commensal Neisseria strains are well documented. Recent data now indicate that sequence-related false-negative results can occur in gonococcal NAATs, whereby target sequences either are absent or contain several mutations. In this study, a duplex Neisseria gonorrhoeae real-time polymerase chain reaction (PCR) (NGduplex) assay targeting the gonococcal porA pseudogene and multicopy opa genes was developed. The NGduplex was evaluated by testing 596 clinical specimens, including 292 urogenital specimens and 304 throat swab specimens. The results were compared with those obtained using a consensus reference standard comprising 3 monoplex real-time PCR assays. The results show that the NGduplex assay is highly suitable for routine screening for gonorrhea, providing an overall clinical sensitivity and specificity of 100% and 99.3%, respectively, for both urogenital and throat swab specimens. In addition, the 2-target system of the NGduplex assay decreases the potential for sequence-related false-negative results and can provide simultaneous confirmation of positive results. (C) 2008 Elsevier Inc. All rights reserved.
Formatted abstract
Cross-reactions of gonococcal nucleic acid amplification tests (NAATs) with commensal Neisseria strains are well documented. Recent data now indicate that sequence-related false-negative results can occur in gonococcal NAATs, whereby target sequences either are absent or contain several mutations. In this study, a duplex Neisseria gonorrhoeae real-time polymerase chain reaction (PCR) (NGduplex) assay targeting the gonococcal porA pseudogene and multicopy opa genes was developed. The NGduplex was evaluated by testing 596 clinical specimens, including 292 urogenital specimens and 304 throat swab specimens. The results were compared with those obtained using a consensus reference standard comprising 3 monoplex real-time PCR assays. The results show that the NGduplex assay is highly suitable for routine screening for gonorrhea, providing an overall clinical sensitivity and specificity of 100% and 99.3%, respectively, for both urogenital and throat swab specimens. In addition, the 2-target system of the NGduplex assay decreases the potential for sequence-related false-negative results and can provide simultaneous confirmation of positive results.

Keyword Neisseria gonorrhoeae GC
Real-time PCR
Opa
porA
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: 2009 Higher Education Research Data Collection
Excellence in Research Australia (ERA) - Collection
 
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Citation counts: TR Web of Science Citation Count  Cited 33 times in Thomson Reuters Web of Science Article | Citations
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Created: Thu, 02 Apr 2009, 01:07:01 EST by Lesley Arnicar on behalf of Clinical Medical Virology Centre