title: Evaluation of six commercial nucleic acid amplification tests for detection of Neisseria gonorrhoeae and other Neisseria species.
abstract: Molecular detection of Neisseria gonorrhoeae in extragenital samples may result in false-positive results due to cross-reaction with commensal Neisseria species or Neisseria meningitidis. This study examined 450 characterized clinical culture isolates, comprising 216 N. gonorrhoeae isolates and 234 isolates of nongonococcal Neisseria species (n = 218) and 16 isolates of other closely related bacteria, with six commercial nucleic acid amplification tests (NAATs). The six NAATs tested were Gen-Probe APTIMA COMBO 2 and APTIMA GC, Roche COBAS Amplicor CT/NG and COBAS 4800 CT/NG tests, BD ProbeTec GC Qx amplified DNA assay, and Abbott RealTime CT/NG test. All assays except COBAS Amplicor CT/NG test where four (1.9%) isolates were not detected showed a positive result with all N. gonorrhoeae isolates (n = 216). Among the 234 nongonococcal isolates examined, initial results from all assays displayed some false-positive results due to cross-reactions. Specifically, the COBAS Amplicor and ProbeTec tests showed the highest number of false-positive results, detecting 33 (14.1%) and 26 (11%) nongonococcal Neisseria isolates, respectively. On the first testing, APTIMA COMBO 2, APTIMA GC, Abbott RealTime, and Roche COBAS 4800 showed lower level of cross-reactions with five (2.1%), four (1.7%), two (1%), and two (1%) of the isolates showing low-level positivity, respectively. Upon retesting of these nine nongonococcal isolates using freshly cultured colonies, none were positive by the APTIMA COMBO 2, Abbott RealTime, or COBAS 4800 test. In conclusion, the COBAS Amplicor and ProbeTec tests displayed high number of false-positive results, while the remaining NAATs showed only sporadic low-level false-positive results. Supplementary testing for confirmation of N. gonorrhoeae NAATs remains recommended with all samples tested, in particular those from extragenital sites.
authors.affiliation: Department of Microbiology and Infectious Diseases, The Royal Women's Hospital, Bio 21 Institute, Parkville 3052, Australia. sepehr.tabrizi@thewomens.org.au
authors.forename: Sepehr N
authors.initials: SN
authors.lastname: Tabrizi
authors.forename: Magnus
authors.initials: M
authors.lastname: Unemo
authors.forename: Athena E
authors.initials: AE
authors.lastname: Limnios
authors.forename: Tiffany R
authors.initials: TR
authors.lastname: Hogan
authors.forename: Stig-Ove
authors.initials: SO
authors.lastname: Hjelmevoll
authors.forename: Susanne M
authors.initials: SM
authors.lastname: Garland
authors.forename: John
authors.initials: J
authors.lastname: Tapsall
mesh_terms: D001431:Bacteriological Techniques / Q000379:methods*; D005189:False Positive Reactions; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009343:Neisseria / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D016870:Neisseriaceae Infections / Q000175:diagnosis* / Q000382:microbiology*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity