Evaluation of opa-based real-time PCR for detection of Neisseria gonorrhoeae
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Sepehr N Tabrizi1, Shujun Chen, John Tapsall
1Department of Molecular Microbiology, The Royal Women's Hospital, Carlton, Victoria 3053, Australia. Tabrizi@wch.org.au
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.
