A new confirmatory Neisseria gonorrhoeae real-time PCR assay targeting the porA pseudogene

Whiley, D. M., Buda, P. J., Bayliss, J., Cover, L., Bates, J. and Sloots, T. P. (2004) A new confirmatory Neisseria gonorrhoeae real-time PCR assay targeting the porA pseudogene. European Journal of Clinical Microbiology & Infectious Diseases, 23 9: 705-710. doi:10.1007/s10096-004-1170-0


Author Whiley, D. M.
Buda, P. J.
Bayliss, J.
Cover, L.
Bates, J.
Sloots, T. P.
Title A new confirmatory Neisseria gonorrhoeae real-time PCR assay targeting the porA pseudogene
Journal name European Journal of Clinical Microbiology & Infectious Diseases   Check publisher's open access policy
ISSN 0934-9723
Publication date 2004-09-01
Year available 2004
Sub-type Article (original research)
DOI 10.1007/s10096-004-1170-0
Open Access Status Not yet assessed
Volume 23
Issue 9
Start page 705
End page 710
Total pages 6
Editor I. Braveny
Place of publication Heidleberg, Germany
Publisher Springer-Verlag
Language eng
Subject C1
270303 Virology
730101 Infectious diseases
Abstract The Roche Cobas Amplicor system is widely used for the detection of Neisseria gonorrhoeae but is known to cross react with some commensal Neisseria spp. Therefore, a confirmatory test is required. The most common target for confirmatory tests is the cppB gene of N. gonorrhoeae. However, the cppB gene is also present in other Neisseria spp. and is absent in some N. gonorrhoeae isolates. As a result, laboratories targeting this gene run the risk of obtaining both false-positive and false-negative results. In the study presented here, a newly developed N. gonorrhoeae LightCycler assay (NGpapLC) targeting the N. gonorrhoeae porA pseudogene was tested. The NGpapLC assay was used to test 282 clinical samples, and the results were compared to those obtained using a testing algorithm combining the Cobas Amplicor System (Roche Diagnostics, Sydney, Australia) and an in-house LightCycler assay targeting the cppB gene (cppB-LC). In addition, the specificity of the NGpapLC assay was investigated by testing a broad panel of bacteria including isolates of several Neisseria spp. The NGpapLC assay proved to have comparable clinical sensitivity to the cppB-LC assay. In addition; testing of the bacterial panel showed the NGpapLC assay to be highly specific for N. gonorrhoeae DNA. The results of this study show the NGpapLC assay is a suitable alternative to the cppB-LC assay for confirmation of N. gonorrhoeae-positive results obtained with Cobas Amplicor.
Keyword Neisseria Gonorrhoeae
Real-time Pcr Assay
Pora Pseudogene
Infectious Diseases
Microbiology
Polymerase-chain-reaction
Clinical-samples
Meningitidis
Gene
Q-Index Code C1
Institutional Status UQ

 
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Created: Wed, 15 Aug 2007, 14:22:21 EST