Polymerase chain reaction for the detection of Neisseria gonorrhoeae in clinical samples.

B, Ho; Wang, Feng; Wong, B. K. C.; Egglestone, S. I.

doi:10.1136/jcp.45.5.439

Cited by 94 publications

(48 citation statements)

References 17 publications

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“…The cppB gene has frequently been used as an N. gonorrhoeae-specific target in PCR assays (10), but the specificity for N. gonorrhoeae of the cppB gene has recently been challenged (2,3,15,17,23,29). Although found on the cryptic plasmid pJD1, which is often at higher copy numbers than chromosomal genes, there are some N. gonorrhoeae strains that are cppB negative by PCR due to lack of the plasmid or chromosomal integration (2,3,15,17,23).…”

Section: Discussionmentioning

confidence: 99%

Neisseria Species Identification Assay for the Confirmation of Neisseria gonorrhoeae -Positive Results of the COBAS Amplicor PCR

Mangold

Regner

Tajuddin³

et al. 2007

J Clin Microbiol

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Screening assays for Neisseria gonorrhoeae exhibit low positive predictive values, particularly in low-prevalence populations. A new real-time PCR assay that detects and identifies individual Neisseria spp. using melt curve analysis was compared to two previously published supplementary assays. NsppID, a 16S rRNA real-time PCR/melt curve assay developed to distinguish N. gonorrhoeae from other Neisseria spp., was compared to real-time PCR assays targeting genes reportedly specific for N. gonorrhoeae, the cppB gene and the porA pseudogene. A total of 408 clinical specimens (324 female endocervical swabs and 84 male urine or urogenital swab specimens) were screened using the COBAS Amplicor assay for Chlamydia trachomatis and N. gonorrhoeae (CT/NG) (Roche Diagnostics, Indianapolis, IN) followed by confirmatory testing via real-time PCR. The NsppID assay detected Neisseria spp. in 150/181 COBAS-positive specimens (82%), including six dual infections, and identified N. gonorrhoeae in 102 (56%) specimens. Sixty-nine of 181 (38%) specimens were positive for N. gonorrhoeae by porA pseudogene, and 115/181 (64%) were positive for cppB. However, cppB was also positive in 15% of COBAS-negative specimens, more than either NsppID (4%) or porA pseudogene (2%) assays. The porA pseudogene assay had the highest specificity for both genders but the lowest sensitivity, especially in female specimens. NsppID had a slightly lower specificity but greater sensitivity and overall accuracy. The least desirable confirmatory assay was cppB, due to poor specificity. The NsppID assay is an accurate confirmatory assay for N. gonorrhoeae detection. In addition, the NsppID assay can identify the non-N. gonorrhoeae species responsible for the majority of false-positive results from the COBAS Amplicor CT/NG assay.Neisseria gonorrhoeae is the second most prevalent sexually transmitted bacterium after Chlamydia trachomatis (4). Many N. gonorrhoeae infections are asymptomatic, especially in women, and if left untreated can develop long-term consequences, including chronic pelvic pain, pelvic inflammatory disease, and ascending infection of the fallopian tubes. Accurate diagnosis of symptomatic and asymptomatic infection is required to prevent these complications and to control the transmission of infection.Molecular assays such as the COBAS Amplicor CT/NG have high sensitivity and specificity in detecting both C. trachomatis and N. gonorrhoeae infections, and screening of high-prevalence populations results in positive predictive values (PPVs) of Ͼ80%. However, even with very specific assays, screening of low-prevalence populations lowers the positive predictive value of any test. For N. gonorrhoeae analysis of low-prevalence populations performed by the COBAS Amplicor CT/NG assay (6, 12, 32), unacceptable positive predictive values have been reported, particularly when the optical density (OD) reading is in the gray-zone of Ն0.2 to Ͻ3.5 (6). Repeat testing of the gray-zone and positive specimens has been shown to improve the specificity of the COBA...

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Section: Discussionmentioning

confidence: 99%

Neisseria Species Identification Assay for the Confirmation of Neisseria gonorrhoeae -Positive Results of the COBAS Amplicor PCR

Mangold

Regner

Tajuddin³

et al. 2007

J Clin Microbiol

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“…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).…”

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confidence: 99%

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

et al. 2004

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“…It is proven that clinical isolates of Neisseria subflava and Neisseria cinerea, belonging to the commensal flora of the human respiratory or genital tract, may exhibit cross-reactivity in the AMPLICOR PCR (5,11). Accurate confirmation assays include other commercially available NAATs as well as noncommercial PCRs targeting the ccpB gene on the 2.6-MDa cryptic plasmid or DNA encoding the 16S rRNA (a prototype of the 16S rRNA PCR was developed by Roche Diagnostics Systems, Branchburg, N.J.) (3,(5)(6)(7).…”

mentioning

confidence: 99%

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

et al. 2001

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Polymerase chain reaction for the detection of Neisseria gonorrhoeae in clinical samples.

Cited by 94 publications

References 17 publications

Neisseria Species Identification Assay for the Confirmation of Neisseria gonorrhoeae -Positive Results of the COBAS Amplicor PCR

Neisseria Species Identification Assay for the Confirmation of Neisseria gonorrhoeae -Positive Results of the COBAS Amplicor PCR

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

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

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