Evaluation of the Roche Neisseria gonorrhoeae 16S rRNA PCR for Confirmation of AMPLICOR PCR-Positive Samples and Comparison of Its Diagnostic Performance According to Storage Conditions and Preparation of Endocervical Specimens

Abstract

The AMPLICOR PCR was used to detect Neisseria gonorrhoeae in endocervical specimens. A 16S rRNA PCR performed on N. gonorrhoeae-positive samples showed sensitivities of 73.2, 64.3, and 94.4% for samples treated directly with AMPLICOR lysis buffer, samples suspended in 2-sucrose phosphate, and samples suspended in diluted phosphate-buffered saline, respectively.

Evaluation of an automated extraction method for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae by Cobas Amplicor PCR from different sample materials

“…Samples yielding different results with both methods were re-extracted in parallel on both the MP instrument and by spin columns. Because of earlier studies indicating nonspecific reactions of NG Amplicor primers with some Neisseria species other than gonorrhoeae (such as N. subflava, N. cinera and N. lactamica) [16,17], all samples yielding a result with an OD value greater than 0.2 for NG were also subjected to a NG 16S rDNA confirmation test (Roche; [18,19]). …”

Multicenter Validation of the cppB Gene as a PCR Target for Detection of Neisseria gonorrhoeae

“…Confirmatory tests using the 16S rRNA genes and the cppB gene have been reported (2,5,13,14); however, the cryptic plasmid on which the cppB gene is located is suspected to be missing in some clinical isolates (11). Therefore, we determined the frequency of the cppB gene in well-characterized N. gonorrhoeae strains cultured from STI patients by using realtime PCR technologies as developed in different diagnostic laboratories in The Netherlands (Table 1).…”

Specific andSensitive Detection of Neisseria gonorrhoeae in ClinicalSpecimens by Real-Time PCR

“…The COBAS AMPLICOR test for N. gonorrhoeae (COBAS AMPLICOR CT/NG; Roche Diagnostics Nederland BV, Almere, The Netherlands), for instance, produces false-positive results with certain nonpathogenic Neisseria species (Neisseria subflava and Neisseria cinerea) and lactobacilli, and a subsequent confirmation test is necessary (5,12,15,30,38). CppB-and 16S rRNA gene-based assays are used for confirmation (35); however, about 5% of N. gonorrhoeae strains do not carry the CppB plasmid (5, 7), and not all 16S rRNA-based tests are sensitive and specific enough (12,39).…”