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. 2005 Oct;37(5):364-70. doi: 10.1080/00313020500254552.


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-ABI7500) 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.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Chlamydia Infections / diagnosis
  • Chlamydia trachomatis / classification
  • Chlamydia trachomatis / genetics
  • Chlamydia trachomatis / isolation & purification*
  • DNA Primers / chemistry
  • DNA, Bacterial / analysis
  • Female
  • Gonorrhea / diagnosis
  • Humans
  • Male
  • Middle Aged
  • Neisseria gonorrhoeae / classification
  • Neisseria gonorrhoeae / genetics
  • Neisseria gonorrhoeae / isolation & purification*
  • Polymerase Chain Reaction / methods*
  • Reproducibility of Results
  • Sensitivity and Specificity


  • DNA Primers
  • DNA, Bacterial