title: Purification of DNA eluates from the ProbeTec GC Q(x) assay on BD Viper™ XTR allows for further analysis and confirmation of gonorrhea.
abstract: Low positive predictive values of Neisseria gonorrhoeae nucleic acid amplification tests in low-prevalence populations are a major challenge for accurate diagnostics. It is therefore necessary to verify all positive N. gonorrhoeae results with a different assay and target gene, preferably using the same sample. The BD ProbeTec™ Q(x) Collection Kit for Endocervical or Lesion Specimens, which is recommended for BD Viper™ XTR, is incompatible with other commercial platforms. Therefore, a confirmatory PCR has not been available for samples received on this transport medium. To be able to verify results from these samples with another assay, our objective was to establish a procedure for using the DNA eluates from BD Viper™ XTR for further analysis. DNA eluates from BD Viper™ XTR were collected and analyzed in two in-house confirmatory real-time PCRs targeting the porA pseudogene and the opa multicopy gene. BD Viper™ XTR DNA eluates were analyzed directly and also after purification with the nucleic acid extraction system NucliSENS® easyMag®. Purification of BD Viper™ XTR DNA eluates with the nucleic acid extraction system NucliSENS® easyMag® provided a sensitivity of the in-house PCR comparable to BD Viper™. With the inclusion of two target genes in the confirmatory PCR, specific and reliable verification of results were obtained. This study presents a simple, inexpensive procedure which allows for rapid verification of gonorrhea from samples received on the BD ProbeTec™ Q(x) Collection Kit for Endocervical or Lesion Specimens.
authors.affiliation: Unit of Gene Technology and Department of Microbiology, Division of Diagnostics and Technology, Akershus University Hospital, Sykehusveien 27, 1478, Nordbyhagen, Norway, hesm@ahus.no.
authors.forename: H S
authors.initials: HS
authors.lastname: Tunsjø
authors.forename: E
authors.initials: E
authors.lastname: Smedsrud
authors.forename: P C
authors.initials: PC
authors.lastname: Holm
authors.forename: K
authors.initials: K
authors.lastname: Rokvam
authors.forename: C
authors.initials: C
authors.lastname: Schie
authors.forename: V
authors.initials: V
authors.lastname: Rognlien
authors.forename: I
authors.initials: I
authors.lastname: Augustin
authors.forename: A
authors.initials: A
authors.lastname: Fostervold
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D006069:Gonorrhea / Q000175:diagnosis*; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009344:Neisseria gonorrhoeae / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D018272:Porins / Q000235:genetics; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods*
chemical_list: D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial; D018272:Porins; C057699:porin protein, Neisseria; C090348:Opa protein, Neisseria
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