title: Characteristics of the m2000 automated sample preparation and multiplex real-time PCR system for detection of Chlamydia trachomatis and Neisseria gonorrhoeae.
abstract: We evaluated a new real-time PCR-based prototype assay for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae developed by Abbott Molecular Inc. This assay is designed to be performed on an Abbott m2000 real-time instrument system, which consists of an m2000sp instrument for sample preparation and an m2000rt instrument for real-time PCR amplification and detection. The limit of detection of this prototype assay was determined to be 20 copies of target DNA for both C. trachomatis and N. gonorrhoeae, using serially diluted linearized plasmids. No cross-reactivity could be detected when 55 nongonococcal Neisseria isolates and 3 non-C. trachomatis Chlamydia isolates were tested at 1 million genome equivalents per reaction. Concordance with the Roche Amplicor, BDProbeTec ET, and Gen-Probe APTIMA Combo 2 tests was assessed using unlinked/deidentified surplus clinical specimens previously analyzed with these tests. For C. trachomatis, concordance for positive results ranged from 93.7% to 100%, while concordance for negative results ranged from 98.2% to 100%. For N. gonorrhoeae, concordance for positive and negative results ranged from 91.4% to 100% and 99.3% to 100%, respectively. A workflow analysis of the prototype assay was conducted to obtain information on throughput under laboratory conditions. At 48 samples/run, the time to first result for both C. trachomatis and N. gonorrhoeae was 4.5 h. A total of 135 patient specimens could be analyzed in 8.9 h, with 75 min of hands-on time. This study demonstrated the technical and clinical feasibility of the new Abbott real-time PCR C. trachomatis/N. gonorrhoeae assay.
authors.affiliation: Abbott Molecular Inc., Des Plaines, IL 60018, USA.
authors.forename: R
authors.initials: R
authors.lastname: Marshall
authors.forename: M
authors.initials: M
authors.lastname: Chernesky
authors.forename: D
authors.initials: D
authors.lastname: Jang
authors.forename: E W
authors.initials: EW
authors.lastname: Hook
authors.forename: C P
authors.initials: CP
authors.lastname: Cartwright
authors.forename: B
authors.initials: B
authors.lastname: Howell-Adams
authors.forename: S
authors.initials: S
authors.lastname: Ho
authors.forename: J
authors.initials: J
authors.lastname: Welk
authors.forename: J
authors.initials: J
authors.lastname: Lai-Zhang
authors.forename: J
authors.initials: J
authors.lastname: Brashear
authors.forename: B
authors.initials: B
authors.lastname: Diedrich
authors.forename: K
authors.initials: K
authors.lastname: Otis
authors.forename: E
authors.initials: E
authors.lastname: Webb
authors.forename: J
authors.initials: J
authors.lastname: Robinson
authors.forename: H
authors.initials: H
authors.lastname: Yu
mesh_terms: D001331:Automation; D002690:Chlamydia Infections / Q000175:diagnosis* / Q000382:microbiology; D002692:Chlamydia trachomatis / Q000235:genetics / Q000302:isolation & purification*; D005260:Female; D052776:Female Urogenital Diseases / Q000175:diagnosis / Q000382:microbiology; D006069:Gonorrhea / Q000175:diagnosis* / Q000382:microbiology; D006801:Humans; D008280:Magnetics; D008297:Male; D052801:Male Urogenital Diseases / Q000175:diagnosis / Q000382:microbiology; D009344:Neisseria gonorrhoeae / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods
chemical_list: D011933:Reagent Kits, Diagnostic