Evaluation of opa-based real-time PCR for detection of Neisseria gonorrhoeae

Sex Transm Dis. 2005 Mar;32(3):199-202. doi: 10.1097/01.olq.0000154495.24519.bf.

Abstract

Objectives: Detection of Neisseria gonorrhoeae by commercial and in-house-based assays has been hampered by false-positive and false-negative results. The current study describes a sensitive and specific real-time 5'-nuclease PCR assay targeting a 90-bp region of the multicopy opa gene.

Goal: To evaluate the sensitivity and specificity of this assay in detection of gonococcus.

Study: Sensitivity and specificity were determined by testing a panel of 173 microorganisms. In addition, 135 clinical samples previously evaluated by 4 nucleic acid amplification methods were also tested.

Results: A sensitivity of 1 copy per reaction was achieved. Positive results were only obtained for N gonorrhoeae strains including 20 cppB-negative strains. Overall, 134 of 135 clinical sample results agreed with the consensus nucleic amplification methods.

Conclusion: This study demonstrates opa-based target can be used as an accurate and rapid PCR assay for the detection of N gonorrhoeae in clinical specimens.

Publication types

  • Evaluation Study

MeSH terms

  • Bacterial Outer Membrane Proteins / analysis
  • Bacterial Outer Membrane Proteins / genetics*
  • DNA Primers
  • DNA, Bacterial / analysis
  • Gonorrhea / epidemiology
  • Gonorrhea / microbiology
  • Gonorrhea / prevention & control*
  • Humans
  • Neisseria gonorrhoeae / genetics
  • Neisseria gonorrhoeae / isolation & purification*
  • New South Wales / epidemiology
  • Polymerase Chain Reaction / methods
  • Predictive Value of Tests
  • Sensitivity and Specificity
  • Victoria / epidemiology

Substances

  • Bacterial Outer Membrane Proteins
  • DNA Primers
  • DNA, Bacterial
  • Opa protein, Neisseria