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


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


METHODS
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.


CONCLUSIONS
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.
Tabrizi
Sepehr N
SN
Department of Molecular Microbiology, The Royal Women's Hospital, Carlton, Victoria 3053, Australia. Tabrizi@wch.org.au
Chen
Shujun
S
Tapsall
John
J
Garland
Suzanne M
SM
D001425:Bacterial Outer Membrane Proteins / Q000032:analysis / Q000235:genetics*; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis; D006069:Gonorrhea / Q000453:epidemiology / Q000382:microbiology / Q000517:prevention & control*; D006801:Humans; D009344:Neisseria gonorrhoeae / Q000235:genetics / Q000302:isolation & purification*; D009517:New South Wales / Q000453:epidemiology; D016133:Polymerase Chain Reaction / Q000379:methods; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity; D014739:Victoria / Q000453:epidemiology
D001425:Bacterial Outer Membrane Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; C090348:Opa protein, Neisseria