A comparison of three real-time PCR assays for the confirmation of Neisseria gonorrhoeae following detection of N. gonorrhoeae using Roche COBAS AMPLICOR

Abstract

Three different real-time PCR assays were evaluated as confirmatory tests for Neisseria gonorrhoeae after initial screening using the COBAS AMPLICOR Chlamydia trachomatis and N. gonorrhoeae duplex assay. The target genes used for the confirmation were the gyr, cppB and 16S rRNA genes. Analytical specificity was determined by testing 60 strains belonging to different bacterial species and/or serogroups. The primers chosen from the 16S rRNA gene for confirmation of N. gonorrhoeae were highly specific, showed no cross-reactivity with other bacteria included in the study, and had an analytical sensitivity of 1 CFU. Of 192 clinical specimens that were positive for N. gonorrhoeae according to the COBAS AMPLICOR assay, 42 were confirmed as positive using the 16S rRNA gene target, 26 were confirmed using the cppB target, and 30 were confirmed using the gyr target. It was concluded that the real-time PCR assay targeting the 16S rRNA gene is a useful confirmatory assay to complement the COBAS AMPLICOR screening test for N. gonorrhoeae.

Process Evaluation of a Fully Automated Molecular Diagnostics System

“…Several partially or fully automated molecular diagnostic systems are currently available, such as the Roche COBAS AMPLICOR System (Roche Diagnostics, Indianapolis, IN), 10,11 the Abbott m2000 (Abbott Diagnostics, Abbott Park, IL), 9 the GenProbe Tigris DTS (GenProbe, San Diego, CA), 9 and the BD Viper System with XTR (BD Diagnostic Systems, Sparks, MD). 9 Because limited information is available regarding the process efficiency, skill level, and task intensity necessary to operate these instruments, we undertook to analyze the BD Viper System with XTR, which is designed for unattended extraction, amplification, and detection of CT and GC in urine or swabs of vaginal and urethral mucosa.…”

Characterization of Gibberellin Biosynthetic Gene Cluster from Fusarium proliferatum

“…In this report, we describe the isolation and functional characterization of the GA biosynthesis genes cluster in GA-producing F. proliferatum KGL0401. Real-time gene expression studies [6,10,21] of GA biosynthesis genes in regard to high (carbon : nitrogen = 0.5 M : 0.17 M), and low (carbon : nitrogen = 0.25 M : 0.47 M) GA production revealed significant differences in their regulation.…”

An amplification-free, 16S rRNA test for Neisseria gonorrhoeae in urine

“…The complementary PNA probe sequence with PEG linker for detecting N. gonorrhoeae 16S rRNA consisted of NH 2 -(CH 2 CH 2 OCH 2 CH 2 -OCH 2 CO) 6 -TTG CCA ATA TCG GCG GCC. 13 In order to prepare microspheres to be conjugated with PNA, one μL of 820 nm-diameter carboxylic group-functionalized polystyrene microspheres, at a concentration of ∼3.25 × 10 11 mL −1 , was suspended and washed three times in 100 mM MES buffer (pH 4.5). For each wash step, after centrifugation at 14000 rpm for 15 minutes, the sedimented pellet microspheres were resuspended in fresh MES buffer.…”