Comparison of three in-house multiplex PCR assays for the detection of Neisseria gonorrhoeae and Chlamydia trachomatis using real-time and conventional detection methodologies

Whiley, David M. and Sloots, Theo P. (2005) Comparison of three in-house multiplex PCR assays for the detection of Neisseria gonorrhoeae and Chlamydia trachomatis using real-time and conventional detection methodologies. Pathology, 37 5: 364-370. doi:10.1080/00313020500254552


Author Whiley, David M.
Sloots, Theo P.
Title Comparison of three in-house multiplex PCR assays for the detection of Neisseria gonorrhoeae and Chlamydia trachomatis using real-time and conventional detection methodologies
Journal name Pathology   Check publisher's open access policy
ISSN 0031-3025
Publication date 2005
Sub-type Article (original research)
DOI 10.1080/00313020500254552
Volume 37
Issue 5
Start page 364
End page 370
Total pages 7
Editor C. S. Lee
Place of publication Australia
Publisher Taylor & Francis
Collection year 2005
Language eng
Subject C1
270303 Virology
730101 Infectious diseases
Formatted abstract
Aims:
To develop and evaluate multiplex PCR assays for Chlamydia trachomatis and Neisseria gonorrhoeae, using real-time and conventional PCR detection methodologies.

Methods:

Two real-time multiplex PCR assays, using nuclease (TaqMan-AB17500) and hybridisation (LightCycler) probe formats, and a third assay using conventional PCR with solid-phase hybridisation and colour detection, were developed. The porA pseudogene was targeted for N. gonorrhoeae, and the major outer membrane protein gene for C. trachomatis. A total of 145 urogenital specimens were tested in all assays, and the results were compared with the Roche Cobas Amplicor assay.

Results:

There was little difference in clinical sensitivity and specificity, result discrimination and test cost for the three in-house assays. Our results showed that competitive inhibition of the PCR occurred in some samples that were positive for both organisms.

Conclusions:
These results highlight the suitability and versatility of three multiplex PCR methods for the detection of C. trachomatis and N. gonorrhoeae.
Keyword Gonorrhoea
Chlamydia
Real-time
Pcr
Multiplex
Pathology
Amplification Methods
Specimens
Amplicor
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

 
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Created: Wed, 15 Aug 2007, 06:31:38 EST